Flura-seq for Evaluating the Effects of Different Hyperthermic Intraperitoneal Chemotherapy Regimens on the Transcriptome of Pseudomyxoma Peritonei: Complete Statistical Analysis

1. Trial at a Glance

This page covers the registry results for Flura-seq for Evaluating the Effects of Different Hyperthermic Intraperitoneal Chemotherapy Regimens on the Transcriptome of Pseudomyxoma Peritonei (NCT06839378), a randomized na study. The primary outcome is Changes in RNA in Tumor Tissues Before and After HIPEC.

2. Clinical Question

The study is intended to examine the effect of the regimens on newly generated tumor RNA using Flura-seq. Its primary outcome is a measure of transcriptomic change, not a direct clinical outcome such as survival, symptoms, or tumor control. The registry describes more changes in newly generated RNA as indicating a greater impact on the tumor transcriptome; it presents that interpretation as a possible indicator of efficacy, not as a validated clinical measure.

The record also includes a secondary comparison of transcriptomic changes detected by Flura-seq and bulk RNA-seq. That comparison concerns measurement approaches rather than the randomized treatment contrast. Both questions are relevant to understanding how the treatment regimens and the RNA-sequencing approach relate to observed molecular changes, but they answer different questions.

3. Trial Design

The registry describes a randomized, open-label, parallel-group study. Enrollment is listed as 36, with 18 participants in the cisplatin group and 18 in the cisplatin plus docetaxel group. The study phase is listed as NA. The sponsor is Beijing Tsinghua Chang Gung Hospital.

Both groups receive HIPEC after cytoreductive surgery. The cisplatin group receives cisplatin alone; the other group receives cisplatin and docetaxel. Each regimen is described as a perfusion in heated saline at 43 ℃, with a flow rate of 400 ml/min for 60 minutes. The record separately lists an intervention of intravenous 5-FU at 400 mg/m2. How that intervention relates to the two HIPEC groups or to the timing of the tissue collection is not reported in the registry record.

Tumor tissue was collected before and after HIPEC for RNA sequencing. The stated collection window runs from initiation of cytoreductive surgery to completion of HIPEC; the registry describes CRS as taking 4-8 hours and HIPEC as taking 1 hour. This makes the primary outcome a within-tumor pre/post measurement summarized for comparison between treatment groups. The record does not give further details on how tissue samples were selected, processed, or matched to individual participants.

The registry lists the study as completed, with a start date of 2025-02-17 and primary completion date of 2025-08-06. Those status details describe the registry record and do not, by themselves, establish the completeness of outcome data or the conduct of every planned analysis.

4. Participant Flow and Accounting

The registry reports how participants moved through each stage of the study. All counts below are exactly as posted; missing counts are not inferred from other stages.

Before assignment: After providing informed consent, 36 enrolled participants entered a screening phase to confirm eligibility based on predefined inclusion/exclusion criteria. All 36 participants successfully completed screening and proceeded to randomization. No participants were excluded or withdrew during this phase. Using a randomization table provided by the Beijing Tsinghua Changgung Hospital Clinical Data Management Center, the 36 participants were randomly assigned to two groups (18 participants per group

Overall Study

5. Baseline Characteristics

The table reproduces the baseline characteristics posted in the registry, which describe the participants analysed at baseline and help show whether the randomized groups were comparable.

Population: All 36 participants who were randomized and assigned to the two study arms (18 per group) are included in the baseline analysis. There was no discrepancy between the number of participants enrolled, randomized, and included in the baseline characteristics analysis. No participants were excluded post-randomization prior to baseline assessment.

6. Primary Endpoint

The registry lists the primary outcome as Changes in RNA in Tumor Tissues Before and After HIPEC, measured over From the initiation of Cytoreductive Surgery (CRS) to the completion of Hyperthermic Intraperitoneal Chemotherapy (HIPEC). CRS takes 4-8 hours, HIPEC takes 1 hour. Pre and post-HIPEC tumor tissues were collected for RNA-sequencing..

The primary endpoint is titled “Changes in RNA in Tumor Tissues Before and After HIPEC.” It measures the reported number of changed RNA measurements, with results expressed as a mean and standard deviation. The tissue samples were collected before and after HIPEC, and the registry frames the main comparison as the amount of differentially expressed genes and the magnitude of change between the cisplatin and cisplatin plus docetaxel groups.

This molecular outcome is used to describe how much the tumor transcriptome changes during treatment. A larger count of changed newly generated RNA could indicate a greater transcriptomic impact, according to the registry’s stated rationale. However, a transcriptomic change is not itself a patient-centered clinical outcome, and its clinical meaning cannot be inferred from the count alone.

The registry says results are to be summarized as mean ± standard deviation and analyzed using a t-test or rank-sum test, with an unpaired t-test for the between-group comparison. It states that differences are considered statistically significant when P < 0.05. The record does not provide a detailed operational definition of a changed RNA measurement, the threshold for classifying genes as differentially expressed, or a complete description of how the pre- and post-HIPEC measurements contribute to the reported group-level count.

7. Results Posted in the Registry

• Experimental : Cisplatin Group: mean 1,413 versus 1,927 on control; difference -514.00 (95% CI -1,125 to 97.38), standardized effect d = -0.55; z-test p = 0.099.

8. Secondary Efficacy Endpoints

The registry also posts results for secondary outcomes. For each outcome below, the endpoint type, the test applied to the posted summary statistics, the result and its interpretation are given in turn. Secondary endpoints are usually not adjusted for multiple testing, so they support rather than confirm the primary finding.

Comparison of Transcriptomic Changes Detected by Flura-seq vs. Bulk RNA-seq

• Endpoint type: Continuous: a mean value per participant, compared as a difference in means.
• Test: Unadjusted two-sample z-test for the difference in means, using the posted standard deviations.
• Result:
  • Transcriptomic Changes Detected by Bulk RNA-seq: mean 340.00 versus 1,690 on control; difference -1,350 (95% CI -1,672 to -1,028), standardized effect d = -1.93; z-test p = <0.001.
• Interpretation:
  • The 95% confidence interval excludes no difference, so the mean is statistically lower for Transcriptomic Changes Detected by Bulk RNA-seq than for the control. This is an unadjusted, secondary comparison, so it is suggestive rather than confirmatory.

The secondary outcome compares transcriptomic changes detected by Flura-seq with those detected by bulk RNA-seq. The posted result favors a larger detected count with Flura-seq, and the reported interval and unadjusted p-value indicate a clear difference between the two measurement approaches in this dataset.

This finding is related to the primary result because the treatment comparison relies on measuring RNA changes in tumor tissue. A method that detects a different amount of transcriptomic change may affect how the primary biological signal is described. However, the secondary comparison does not test whether adding docetaxel changes the response to HIPEC. It compares measurement approaches and should be interpreted separately from the randomized treatment contrast.

The record does not explain whether the method comparison used matched samples, how each method’s results were defined, or how differences in detection thresholds and processing were handled. It also does not describe how the secondary comparison was selected in relation to the primary analysis. These details matter when assessing whether the observed method difference reflects sensitivity, differing analytic definitions, or other factors.

Unadjusted secondary results carry less weight when multiple outcomes or analyses may have been examined, because the chance of seeing an apparently unusual result can increase across repeated testing. The registry lists one secondary outcome and does not report a multiplicity adjustment or a broader analysis plan. The secondary result is therefore useful as a descriptive comparison, but it should not be treated as independent confirmation of the primary treatment finding or as proof of clinical benefit.

Secondary outcomes registered for this trial, for reference (only those above were recalculated):

• Comparison of Transcriptomic Changes Detected by Flura-seq vs. Bulk RNA-seq

9. Statistical Interpretation

10. Statistical Methods Explained

The primary outcome is continuous in the registry’s classification and is summarized as a mean with a standard deviation. A mean describes the average reported count across the group, while the standard deviation describes how spread out individual observations are around that average. The standard deviation is not a confidence interval for the mean, and the two convey different kinds of information.

What does the mean difference represent?

The primary contrast is reported as cisplatin minus cisplatin plus docetaxel. A negative estimate therefore indicates a lower average count in the cisplatin group under that subtraction order. The estimate of -514 is the reported difference in group means, not a difference calculated from an individual participant’s paired pre- and post-treatment values. The record does not provide enough detail to reconstruct the participant-level outcome or the aggregation of the tissue measurements.

What does a confidence interval add?

A confidence interval describes statistical uncertainty around an estimate under the assumptions of the method used to calculate it. A narrower interval indicates greater precision; a wider interval leaves more uncertainty about the size and direction of the underlying difference. For the primary contrast, the interval extends from -1125 to 97.38 and includes zero. That is consistent with the unadjusted p-value of 0.099 and indicates that the posted data do not clearly distinguish the group means using the stated significance threshold.

What do the p-values say?

A p-value measures how unusual data at least as inconsistent with a specified null hypothesis would be if that hypothesis and the statistical model were true. It is not the probability that the null hypothesis is true, and it does not measure clinical importance. The record states that P < 0.05 is the threshold for statistical significance. The primary p-value is 0.099; the secondary method comparison is reported as p < 0.001. These are unadjusted results, and the registry does not report a multiplicity procedure.

Why might a rank-sum test be considered?

As a general statistical point, a rank-sum test compares the ordering of observations and can be considered when a distribution is poorly suited to a mean-based test. It does not test the same quantity as a comparison of means in every setting. The registry mentions either a t-test or rank-sum test for the primary outcome and an unpaired t-test for the between-group comparison, but it does not state which test produced the posted primary p-value. The outcome distribution and test-selection rationale are not reported in the registry record.

Why does over-dispersion matter?

As a general explanation, over-dispersion occurs when variability is greater than a model’s assumptions allow. If counts vary substantially across observations, a model that assumes less variability can understate uncertainty and yield intervals or p-values that are too optimistic. The posted endpoint is a count-like measure reported with means and standard deviations. The registry does not report a distributional assessment or a count-model analysis, so the appropriateness of any particular distributional model cannot be judged from the record.

The randomized parallel design supports a comparison between the assigned regimens, but the record’s statistical description is brief. It does not report adjustment variables, a prespecified analysis population, a missing-data strategy, or detailed procedures for handling the pre/post tissue measurements. Those omissions limit independent assessment of whether the posted summaries correspond to the intended analysis plan.

11. Confidence Intervals and Precision

The primary estimated mean difference is -514, with a 95% confidence interval from -1125 to 97.38. The interval spans zero and includes both a sizable negative contrast and a modest positive value. This means the observed estimate favors a higher average count in the cisplatin plus docetaxel group, but the uncertainty does not rule out a difference in the opposite direction. The result is therefore not precise enough to establish the direction of the underlying group difference with confidence.

The standard deviations are substantial relative to the means: 876 in the cisplatin group and 992 in the combination group. This indicates considerable spread in the reported outcome within each group. A group mean can summarize the center of a distribution, but it does not show whether observations are clustered near that mean or vary widely. The registry does not provide distribution plots, participant-level results, or additional summaries that would clarify the shape of the outcome distribution.

The confidence interval is more informative about uncertainty than the p-value alone. A non-significant p-value does not show that the treatment groups are identical, and it does not establish equivalence. Establishing equivalence or non-inferiority would require an appropriate design and a prespecified margin; such a margin is not reported in the registry record.

The secondary method comparison has a confidence interval from -1672 to -1028, which does not include zero. That interval indicates a more clearly separated estimate for the measurement-method contrast than for the primary treatment contrast. It answers a different question, however, and its apparent precision should not be mistaken for precision about the effect of the HIPEC regimens.

12. Planning a Similar Trial: Sample Size

This is a planning illustration, not a re-evaluation of the original trial. It asks: if a new study wanted to detect a difference like the one observed, or a smaller one, how large would it need to be? It is deliberately not “observed power” of the finished trial, which is not a meaningful quantity.

The planning figures are participants per arm, not total event counts. They are illustrations based on the observed effect, not the trial’s original design calculation. The registry record does not report the original sample-size justification or its assumptions.

The planning assumptions are a two-sided significance level of 0.05, equal allocation between the two arms, and a normal approximation. Each planning row considers an effect equal to a stated fraction of the observed effect. The 80% power column gives the estimated participants per arm under those assumptions for an analysis to have an 80% chance of detecting the specified effect if it were present. The 90% power column gives the corresponding planning target for a 90% chance.

The rows based on a smaller fraction of the observed effect require more participants per arm. This is a general feature of power planning: smaller differences are harder to distinguish from random variation, so more information is needed to detect them at the same significance level and power target. The observed groups are smaller than some of the planning illustrations, but these calculations should not be read as a retrospective measure of whether the study was adequately powered.

This is not observed power. The planning figures depend on the observed effect, which is uncertain, and are not evidence about the probability that the study’s conclusion is correct. They are best read as hypothetical planning scenarios under the listed assumptions.

13. How the Sample Size Is Derived

Power is the probability that a study of a given size rejects the null hypothesis when a specific true effect exists. It depends on four things: the effect to detect, the variability of the outcome, the significance level (here a two-sided α of 0.05, so z_1−α/2 = 1.960) and the desired power (z_1−β = 0.842 for 80% and 1.282 for 90%).

Formula

n is the number of participants in each arm, Δ is the difference in means to detect and σ is the common standard deviation.

Worked calculation for this trial

1. Observed difference Δ = -514.000; pooled standard deviation σ = 935.799.
2. n = 2 × (1.96 + 0.842)² × 935.799² / -514.000² = 52.0, rounded up to 53 participants per arm for 80% power.

The smaller-effect rows in the table above repeat the same calculation with the effect multiplied by 0.75 and 0.5 (for ratios, applied to the logarithm of the ratio).

R code

alpha <- 0.05
z_a   <- qnorm(1 - alpha / 2)          # two-sided
delta <- -514                      # observed difference in means
sigma <- 935.79912374398                     # pooled standard deviation
n_per_arm <- function(power, frac = 1) {
  ceiling(round(2 * (z_a + qnorm(power))^2 * sigma^2 / (delta * frac)^2, 6))
}
for (f in c(1, 0.75, 0.5))
  cat(sprintf("%3.0f%% of observed effect: n per arm = %s (80%% power), %s (90%% power)\n", 100 * f,
              format(n_per_arm(0.80, f), big.mark = ","), format(n_per_arm(0.90, f), big.mark = ",")))

# Check against R's built-in function (t-based, so very slightly larger)
cat("power.t.test:", round(power.t.test(delta = delta, sd = sigma, power = 0.80)$n, 1), "\n")

# Simulation: exact two-sample t-test using simulated means and variances
set.seed(6839378)
n    <- n_per_arm(0.80)
reps <- 20000
m_diff <- rnorm(reps, delta, sigma * sqrt(2 / n))             # difference in sample means
s2     <- sigma^2 * rchisq(reps, 2 * (n - 1)) / (2 * (n - 1))   # pooled sample variance
tstat  <- m_diff / sqrt(s2 * 2 / n)
cat(sprintf("Simulated power at n = %s per arm: %.3f (%d replicates)\n", format(n, big.mark = ","), mean(abs(tstat) > qt(0.975, 2 * (n - 1))), reps))

Output from running the code

100% of observed effect: n per arm = 53 (80% power), 70 (90% power)
 75% of observed effect: n per arm = 93 (80% power), 124 (90% power)
 50% of observed effect: n per arm = 209 (80% power), 279 (90% power)
power.t.test: 53 
Simulated power at n = 53 per arm: 0.801 (20000 replicates)

The simulation draws many sets of group means and variances at the planned sample size and counts how often the two-sample t-test rejects the null. It should land close to the target power; at very small required sample sizes it can fall below it, because the normal approximation is rough there.

14. Multiplicity, Missing Data and Analysis Populations

The registry describes one primary endpoint and one secondary outcome. It states that statistical significance is defined as P < 0.05 and reports unadjusted p-values for the posted comparisons. No multiplicity adjustment is reported in the registry record. When several outcomes, contrasts, or analytic choices are examined, the chance of at least one apparently significant result can increase; adjustment procedures or a prespecified testing hierarchy can address that concern. Without a reported strategy, the secondary result should be interpreted cautiously, and the family of analyses relevant to the reported p-values is unclear.

The registry outcome description mentions a t-test or rank-sum test and says that the between-group comparison uses an unpaired t-test. It does not identify which test was used for the posted primary result, whether the analysis was adjusted for other variables, or whether the test choice was made before outcomes were examined. The number of reported analyses is listed as 0, and the reported-analyses list is empty. This does not supply a fuller analysis plan or clarify the provenance of the posted summary statistics.

Missing-data handling is not reported in the registry record. There is no description of whether tissue samples or RNA measurements were unavailable, how incomplete pre/post pairs were treated, or whether any imputation was performed. The analysis population is also not reported: the record does not state whether results include all randomized participants, only participants with evaluable tissue, or another defined group.

The registry lists enrollment and group counts, but it does not explain participant flow through surgery, tissue collection, sequencing, and analysis. Without those details, it is not possible to determine whether the reported groups represent all randomized participants or whether exclusions could have affected the comparison. These reporting gaps do not establish that data were missing or that analyses were biased; they leave those questions unresolved.

15. Safety Results

The registry reports serious adverse events as affected participants divided by participants at risk.

Deaths

Other (non-serious) adverse events

Participants with at least one non-serious event at or above the registry's 0% reporting threshold.

Most frequent serious adverse event terms

Serious events with the highest arm-level rate: venous thromboembolism occurrence (time, symptoms and signs, ultrasound/cta, measures, prognosis) (0.0%), symptomatic deep - vein thrombosis (0.0%), pulmonary embolism (0.0%), post - operative gastrointestinal fistula (0.0%), bone marrow suppression (0.0%).

The registry reports zero serious adverse events in each group among 18 participants at risk per group, and zero deaths in both groups. The listed serious-event categories—including venous thromboembolism, symptomatic deep-vein thrombosis, pulmonary embolism, postoperative gastrointestinal fistula, bone marrow suppression, and intestinal obstruction—are each reported as 0.0%.

These figures describe the recorded safety results during the period covered by the registry record. They do not establish that no adverse effects occurred: the record lists no participants at risk for other adverse events, so non-serious event results are not reported. The record also does not provide detailed safety follow-up duration, event ascertainment procedures, or explanations of how events were attributed to treatment.

With limited group sizes, an absence of recorded serious events should be read as a description of what was reported, not as evidence that the regimens carry no serious risk. The safety figures should also be considered separately from the molecular endpoint, which addresses transcriptomic changes rather than adverse events.

16. What the Result Does, and Does Not, Mean

The primary result describes a difference in the average number of changed RNA measurements between the two HIPEC groups. The point estimate is in the direction of a higher count in the cisplatin plus docetaxel group, but its confidence interval includes zero and the unadjusted p-value does not meet the registry’s stated significance threshold. The data therefore do not provide a precise estimate of the direction or size of the treatment-group difference.

That conclusion is about the measured transcriptomic endpoint. It does not establish that either regimen improves survival, symptoms, quality of life, or tumor control. The registry’s rationale is that more newly generated RNA changes may indicate greater transcriptomic impact and possibly better efficacy. That proposed link is not demonstrated by the primary result alone, and molecular change should not be equated with clinical benefit.

The method comparison suggests that the two sequencing approaches do not yield the same amount of detected change in the posted analysis. This can matter when interpreting molecular endpoints, but it does not establish that one method is more clinically valid or that results from one method predict patient outcomes better. Such conclusions would require validation not reported in the registry record.

The completed status and the presence of posted outcome summaries show that results have been entered in the registry. They do not resolve uncertainties about the analysis population, missing tissue data, outcome definitions, or the statistical plan. Overall, the findings are best viewed as a small randomized study’s reported molecular comparisons, with uncertainty around the treatment contrast and limited information for drawing broader clinical conclusions.

17. Why This Trial Matters Statistically

This study raises a statistical question that is important in molecular oncology research: whether a treatment-related change can be measured reliably in tumor tissue, and whether different sequencing approaches describe that change similarly. Those questions are distinct. A randomized comparison of HIPEC regimens addresses treatment allocation, while a comparison of Flura-seq and bulk RNA-seq addresses measurement.

The primary endpoint also illustrates the difference between a biologically interesting signal and a clinically meaningful outcome. A count of changed RNA measurements can characterize transcriptomic activity, but interpreting it as evidence of efficacy requires a demonstrated relationship with outcomes that matter to patients. The registry presents that relationship as a possibility, not an established result.

The uncertainty around the primary estimate is statistically important. The point estimate alone suggests a directional difference, but the interval includes zero and the p-value does not reach the stated threshold. This is a reminder that a non-significant result is not proof of no difference, while a directional estimate is not proof of a treatment effect.

Finally, the record illustrates why transparent analysis reporting matters. Details about sample availability, endpoint construction, test selection, multiplicity, and the analysis population help readers distinguish a robust planned comparison from an exploratory one. Such information is particularly useful when molecular endpoints involve multiple samples, genes, or measurement methods.

18. Limitations

• The posted summary statistics are the only data these results rest on; patient-level data, full protocols and statistical analysis plans are not available.
• Recalculated results are simple, unadjusted calculations from posted summaries. They do not replace the sponsor's prespecified analysis.

The registry does not give an operational definition of “changed RNA” or specify the threshold used to identify differentially expressed genes. Without those details, it is difficult to assess the biological and statistical interpretation of the reported count. The record also does not describe the processing of pre- and post-HIPEC specimens, the handling of measurements within a participant, or whether tissue sampling was standardized across groups.

The primary outcome is described as a mean number of changed RNA measurements with standard deviations, but no distributional assessment is reported. The analysis description mentions more than one possible test and does not identify which test generated the posted result. Details about adjustments, missing samples, and the analysis population are also not reported in the registry record.

The record lists intravenous 5-FU at 400 mg/m2 as an intervention, while the two arm descriptions focus on the HIPEC regimens. Its role in the treatment sequence or its relation to the comparison is not explained. The registry also does not describe blinding of outcome assessment; masking is listed as none, and RNA analysis procedures are not detailed.

The primary endpoint is a molecular surrogate-like measure, and the registry does not report clinical outcomes that would establish whether transcriptomic differences correspond to patient benefit. The small group sizes and wide primary confidence interval leave substantial uncertainty about the treatment contrast. No original sample-size justification is reported, and the planning illustrations based on the observed effect cannot replace it.

19. Related Tutorials

20. Related Statistical Calculators

21. Sources

• ClinicalTrials.gov: Flura-seq for Evaluating the Effects of Different Hyperthermic Intraperitoneal Chemotherapy Regimens on the Transcriptome of Pseudomyxoma Peritonei — NCT06839378.
• Registry record last updated 2026-01-06; status last verified 2025-12.
• Results first posted 2026-01-06.
• Data retrieved from ClinicalTrials.gov on 2026-11-09.
• Group sizes, means, event counts and sponsor-reported p-values are reproduced as posted. Effect estimates, confidence intervals and p-values not attributed to the sponsor are independently calculated from the posted summary statistics.

22. Record Summary

NCT06839378 is a completed, randomized, open-label, parallel-group study sponsored by Beijing Tsinghua Chang Gung Hospital. It enrolled 36 participants with pseudomyxoma peritonei to compare cisplatin HIPEC with cisplatin plus docetaxel HIPEC after cytoreductive surgery.

The primary outcome measures changes in RNA in tumor tissue before and after HIPEC. The reported mean difference is -514 for cisplatin relative to the combination group, with a 95% confidence interval from -1125 to 97.38 and an unadjusted p-value of 0.099. The secondary comparison concerns transcriptomic changes detected by Flura-seq and bulk RNA-seq.

The registry reports no serious adverse events or deaths in either group. It does not report the original sample-size assumptions, missing-data handling, analysis populations, or multiplicity procedures. The findings are molecular comparisons and do not, on their own, establish clinical benefit.