Objective To investigate the effects of ischemia time and storage periods on RNA quality in fresh-frozen breast cancer(BC)and esophageal cancer(EC)tissue samples in order to establish evidence-based protocols for biobank sample management.Methods The tumor(T)and paired normal(N)tissue samples from 6 cases of BC and 6 cases of EC were collected and cryopreserved in Biobank,Shantou Central Hospital.Mirror paraffin-embedded tissues were simultaneously prepared into sections for morphological analysis.The samples were divided into two groups of<15 min and 15-30 min according to ischemia time,and RNA quality was analyzed at 4 storage periods of 8-10 months(T1),14-16 months(T2),26-28 months(T3)and 38-40 months(T4).Results In 96 analyzed samples,93.8%(90/96)exhibited high quality(RIN≥6),with 89.6%(43/48)in BC and 97.9%(47/48)in EC.Significant differences in RIN were observed between BC group and EC group(8.050 vs.8.600,P=0.009).In EC group,RIN value was significantly negatively correlated with RNA yield(P<0.001).Moreover,RIN values of tumor-normal pairs exhibited markedly significant differences(7.550 vs.9.000,P<0.001).In contrast,no significant difference was detected in BC group(8.200 vs.7.700,P=0.348).Statistical analysis showed that RIN value was positively correlated with 28S/18S(P<0.001),but had no correlation with tumor content(P=0.676)and necrotic content(P=0.055).Neither ischemia time(<15 min vs.15-30 min:8.200 vs.8.300,P=0.932)nor storage periods(T1-T4:8.400,7.700,8.450,8.600,P=0.163)compromised RNA quality.Conclusion Organ origin and tissue type could influence RNA quality of fresh-frozen tissue samples.However,limited ischemia time(≤30 min)and long-term storage period(38-40 months)do not adversely affect RNA quality in fresh-frozen breast cancer and esophageal cancer tissue samples.

In view of the characteristics of H5N6 subtype avian influenza virus(AIV)that it has both high pathogenicity and the risk of cross-species transmission,posing a serious threat to the poultry farming industry and public health security,in order to effectively prevent and control the spread of H5N6 avian influenza,a rapid,sensitive and specific detection technolo-gy was established in this study.The specific monoclonal antibodies against the neuraminidase N6 protein of avian influenza A virus subtype H5N6 were obtained through hybridoma and monoclonal antibody technology.These antibodies were coupled and labeled with carboxyl-functionalized fluorescent quantum dots,along with previously prepared specific antibodies against the hemagglutinin H5 protein.A rapid fluorescence immunochromatographic detection method for the H5N6 subtype of avian influ-enza virus was established according to the principle of double-antibody sandwich immunochromatography.This method a-chieved a detection sensitivity of 1 ng/mL for recombinant hemagglutinin H5 subtype protein and 0.1 ng/mL for recombinant neuraminidase N6 subtype protein.Moreover,the method exhibited no cross-reactivity with other influenza subtypes or patho-gens,such as Newcastle disease(ND),infectious bronchitis(IB),and infectious laryngotracheitis(ILT),thus demonstrating good specificity.The method effectively identified the highly pathogenic avian influenza virus H5 subtype and directly distin-guished the H5N6 subtype with good accuracy.The fluorescent quantum dot immunochromatographic typing detection method established herein met the sensitivity,specificity,and accuracy requirements for H5N6 subtype detection,and can be further used for rapid detection of the H5 and H5N6 subtypes of avian influenza virus.

Objective:To compare the impacts of external fixation of different durations on rehabilitation outcomes after open repair of acute Achilles tendon rupture.Methods:A prospective cohort study was conducted to analyze the clinical data of patients with unilateral acute closed Achilles tendon rupture admitted to Peking University Third Hospital from August 2020 to August 2023. Patients were divided into Group A ( n=96), Group B ( n=347), Group C ( n=346), and Group D ( n=105) based on different postoperative immobilization durations (0, 2, 4 and 6 weeks, respectively). After all the patients received identical open repair procedure, Group A was rehabilitated immediately but the other groups were rehabilitated with the same protocol after removal of the external fixation. Four groups were compared in terms of recovery time of one-leg heel-rise height (OHRH), recovery time of light exercise (LE) in brisk walking and jogging and recovery time of range of motion (ROM). Visual analogue scale (VAS) scores were also compared at 2, 4, 6 and 8 weeks postoperatively. Achilles tendon total rupture score (ATRS) and American Orthopedic Foot & Ankle Society (AOFAS) ankle-hindfoot scores were evaluated at 6, 8, 10, 12, 14 and 16 weeks postoperatively. Complications were recorded. Results:A total of 894 patients including 869 males and 25 females were included, aged 18-60 years [(35.0±6.3)years]. All the patients were followed up for 14-25 months [(19.0±3.0)months]. The recovery time of OHRH in Group A and B was 12.0(12.0, 12.0)weeks and 12.0(10.0, 12.0)weeks, shorter than those in Group C [14.0(14.0, 16.0)weeks] and D [14.0(14.0, 14.0)weeks] ( P<0.05), with no significant difference between Group A and B ( P>0.05) and between Group C and D ( P>0.05). The recovery time of LE in Group A and B was 18.0(18.0, 18.0)weeks and 18.0(16.0, 18.0)weeks, shorter than those in Group C [20.0(20.0, 20.0)weeks] and D [20.0(20.0, 20.0)weeks] ( P<0.05), with no significant difference between Group A and B ( P>0.05) and between Group C and D ( P>0.05). The recovery time of ROM in Group A and B was 6.0(6.0, 6.0)weeks and 6.0(6.0, 6.0)weeks, shorter than those in Group C [8.0(8.0, 10.0)weeks] and D [10.0(10.0, 10.0)weeks)] ( P<0.05), with no significant difference between Group A and B, and between Group C and D ( P>0.05). At 2 weeks postoperatively, the VAS scores were 2.0(1.0, 2.0)points, 2.0(1.0, 2.0)points, and 2.0(1.5, 2.0)points in Group B, C and D, lower than 5.0(5.0, 5.0)points in Group A ( P<0.05), with no significant difference among Group B, C, and D ( P>0.05). At 4 weeks postoperatively, the VAS scores were 1.0(0, 1.0)points, 1.0(0, 1.0)points, and 1.0(0.5, 1.0)points in Group B, C and D, lower than 2.0(1.0, 2.0)points in Group A ( P<0.05), with no significant difference among Group B, C, and D ( P>0.05). At 6 weeks postoperatively, the VAS score was 0(0, 0)points in all the 4 groups, with no significant difference among them ( P>0.05). At 8 weeks postoperatively, the VAS score was 0(0, 0)points, with lower scores in Group A and B than those in Group C and D ( P<0.05) but with no significant difference between Group A and B and between Group C and D ( P>0.05). At 6 weeks postoperatively, the ATRS scores were 52.0(52.0, 53.8)points and 52.0(50.0, 53.0)points in Group A and B, higher than 41.0(38.0, 43.0)points and 19.0(18.0, 20.0)points in Group C and D ( P<0.05), with a higher score in Group C than that in Group D ( P<0.05) but with no significant difference between Group A and B ( P>0.05). At 8 weeks postoperatively, the ATRS scores were 66.0(66.0, 68.0)points in Group A, higher than 63.0(62.0, 64.0)points, 52.0(50.0, 53.0)points, and 39.0(37.0, 40.0)points in Group B, C and D ( P<0.05), with a higher score in Group B than those in Group C and D ( P<0.05) and a higher score in Group C than that in Group D ( P<0.05). At 10 weeks postoperatively, the ATRS score was 75.0(74.0, 76.0)points in Group B, higher than 69.0(69.0, 70.0)points, 72.0(66.0, 74.0)points, and 62.0(58.5, 63.0)points in Group A, C and D ( P<0.05), with higher scores in Group A and C than that in Group D ( P<0.05) but with no significant difference between Group A and C ( P>0.05). At 12 weeks postoperatively, the ATRS score was 84.0(82.0, 85.0)points in Group B, higher than 75.0(75.0, 77.0)points, 79.0(72.0, 81.0)points, and 72.0(71.0, 73.0)points in Group A, C and D ( P<0.05), with higher scores in Group A and C than that in Group D ( P<0.05) but with no significant difference between Group A and C ( P>0.05). At 14 weeks postoperatively, the ATRS score was 87.0(86.0, 87.0)points in Group B, higher than 82.0(82.0, 84.0)points, 83.0(80.0, 85.0)points, and 79.0(77.5, 80.0)points in Group A, C and D ( P<0.05), with higher scores in Group A and C than that in Group D ( P<0.05) but with no significant difference between Group A and C ( P>0.05). At 16 weeks postoperatively, the ATRS scores were 87.0(87.0, 88.0)points and 88.0(87.0, 88.0)points in Group A and B, higher than 86.0(85.0, 87.0)points and 84.0(83.0, 85.0)points in Group C and D ( P<0.05), with a higher score in Group C than that in Group D ( P<0.05) but with no significant difference between Group A and B ( P>0.05). At 6 weeks postoperatively, the AOFAS ankle-hindfoot scores were 94.0(94.0, 95.0)points and 95.0(94.0, 96.0)points in Group A and B, higher than 85.0(83.0, 86.0)points and 74.0(72.0, 75.0)points in Group C and D ( P<0.05), with a higher score in Group C than that in Group D ( P<0.05) but with no significant difference between Group A and B ( P>0.05). At 8 weeks postoperatively, the AOFAS ankle-hindfoot scores were 100.0(99.0, 100.0)points in Group B, higher than 94.0(94.0, 95.0)points, 92.0(90.0, 93.0)points, and 83.0(82.0, 84.0)points in Group A, C and D ( P<0.05), with a higher score in Group A than those in Group C and D ( P<0.05) and a higher score in Group C than that in Group D ( P<0.05). At 10 weeks postoperatively, the AOFAS ankle-hindfoot score was 100.0(100.0, 100.0)points in Group B, higher than 98.0(98.0, 98.0)points, 98.0(96.8, 99.0)points, and 96.0(95.0, 97.0)points in Group A, C and D, with higher scores in Group A and C than that in Group D ( P<0.05) but with no significant difference between Group A and C ( P>0.05). At 12 weeks postoperatively, the AOFAS ankle-hindfoot score was 100.0(100.0, 100.0)points in both Group A and B, with no significant difference between them ( P>0.05), which was higher than 100.0(98.0, 100.0)points and 99.0(98.0, 99.0)points in Group C and D ( P<0.05), with a higher score in Group C than that in Group D ( P<0.05). At 14 and 16 weeks postoperatively, AOFAS ankle-hindfoot score was 100.0(100.0, 100.0)points, with no significant difference among all the groups ( P>0.05). Superficial wound infection occurred in 12 patients [5.2%(5/96) in Group A, 0.6%(2/347) in Group B, 0.6%(2/346) in Group C and 2.9%(3/105) in Group D] ( P<0.01) while rerupture occurred in 16 [9.4%(9/96) in Group A, 1.2% (4/347) in Group B, 0.9%(3/105) in Group C, and 0 patient in Group D] ( P<0.01). Conclusion:For patients with unilateral acute Achilles tendon rupture, two weeks of postoperative external fixation after open repair can shorten the time of returning sports, alleviate pain, and promote functional recovery, without increasing the risk of complications.

Objective:To evaluate the impact of implementing a regional hierarchical medical management model based on the medical internet of things (medical IoT) on the frequency of emergency department visits and hospitalizations, as well as related medical expenses, in patients with chronic obstructive pulmonary disease (COPD).Methods:This retrospective study included COPD patients enrolled in the regional hierarchical medical management system based on Medical IoT across 21 community health service centers in Songjiang District, Shanghai, between July 2017 and May 2018. Utilizing patient data from the year prior to enrollment as the baseline, changes in the number of emergency visits, hospitalizations, and associated medical costs during the first and second years of management were compared. Changes for patients receiving drug treatment were also analyzed.Results:A total of 973 COPD patients were enrolled. The mean age was 75.2±17.0 years, and 64.34% (626/973) were male. Compared to baseline, all COPD patients in the first year of management showed significant reductions: emergency visits decreased by 33.67%, total emergency costs by 45.60%, hospitalizations by 27.15%, and total hospitalization costs by 25.42%. In the second year, reductions were: emergency visits by 28.08%, total emergency costs by 36.10%, hospitalizations by 35.26%, and total hospitalization costs by 18.13% (all P<0.05). Among patients receiving drug therapy, reductions in the first year were: emergency visits by 39.66%, total emergency costs by 47.54%, hospitalizations by 25.19%, and total hospitalization costs by 28.40%. In the second year, reductions were: emergency visits by 46.98%, total emergency costs by 45.99%, hospitalizations by 41.98%, and total hospitalization costs by 24.94% (all P<0.05). No significant differences were observed before and after management for patients without drug treatment. Conclusion:The implementation of the regional hierarchical medical management model based on Medical IoT significantly reduced the frequency of emergency visits and hospitalizations, as well as related costs, for COPD patients.

Background: s/Aims: Sarcomatoid hepatocellular carcinoma (HCC) is a rare histological subtype of HCC characterized by extremely poor prognosis; however, its molecular characterization has not been elucidated. Methods: In this study, we conducted an integrated multiomics study of whole-exome sequencing, RNA-seq, spatial transcriptome, and immunohistochemical analyses of 28 paired sarcomatoid tumor components and conventional HCC components from 10 patients with sarcomatoid HCC, in order to identify frequently altered genes, infer the tumor subclonal architectures, track the genomic evolution, and delineate the transcriptional characteristics of sarcomatoid HCCs. Results: Our results showed that the sarcomatoid HCCs had poor prognosis. The sarcomatoid tumor components and the conventional HCC components were derived from common ancestors, mostly accessing similar mutational processes. Clonal phylogenies demonstrated branched tumor evolution during sarcomatoid HCC development and progression. TP53 mutation commonly occurred at tumor initiation, whereas ARID2 mutation often occurred later. Transcriptome analyses revealed the epithelial–mesenchymal transition (EMT) and hypoxic phenotype in sarcomatoid tumor components, which were confirmed by immunohistochemical staining. Moreover, we identified ARID2 mutations in 70% (7/10) of patients with sarcomatoid HCC but only 1–5% of patients with non-sarcomatoid HCC. Biofunctional investigations revealed that inactivating mutation of ARID2 contributes to HCC growth and metastasis and induces EMT in a hypoxic microenvironment. Conclusions We offer a comprehensive description of the molecular basis for sarcomatoid HCC, and identify genomic alteration (ARID2 mutation) together with the tumor microenvironment (hypoxic microenvironment), that may contribute to the formation of the sarcomatoid tumor component through EMT, leading to sarcomatoid HCC development and progression.

Objective To propose a ventilator failure probability prediction method based on kernel ridge regression(KRR).Methods Firstly,the failure interval data of ventilators was collected and preprocessed to remove outliers.Secondly,the median rank method was used to estimate the failure probability.Finally,using the time data as the feature variable and the failure probability value as the target variable,a KRR model was established and trained by selecting the optimal kernel function and hyperparameter combination from radial basis kernel function,linear kernel function,polynomial kernel function,and S-type kernel function through grid search and cross-validation methods to predict ventilator failures.To verify the performance of the KRR model in predicting ventilator failure probability,it was compared with Weibull and its extended models.Results KRR achieved a coefficient of determination of 0.993 5,a mean squared error of 5.399 5×10-4,a root mean squared error of 0.023 2 and a mean absolute error of 0.018 3,outperforming Weibull and its extended models in prediction accuracy and error control.Conclusion The failure probability prediction method for ventilators based on KRR demonstrates exceptional performance in prediction accuracy and error control,and thus holds great potential for application.[Chinese Medical Equipment Journal,2025,46(5):73-77]

Background: s/Aims: Sarcomatoid hepatocellular carcinoma (HCC) is a rare histological subtype of HCC characterized by extremely poor prognosis; however, its molecular characterization has not been elucidated. Methods: In this study, we conducted an integrated multiomics study of whole-exome sequencing, RNA-seq, spatial transcriptome, and immunohistochemical analyses of 28 paired sarcomatoid tumor components and conventional HCC components from 10 patients with sarcomatoid HCC, in order to identify frequently altered genes, infer the tumor subclonal architectures, track the genomic evolution, and delineate the transcriptional characteristics of sarcomatoid HCCs. Results: Our results showed that the sarcomatoid HCCs had poor prognosis. The sarcomatoid tumor components and the conventional HCC components were derived from common ancestors, mostly accessing similar mutational processes. Clonal phylogenies demonstrated branched tumor evolution during sarcomatoid HCC development and progression. TP53 mutation commonly occurred at tumor initiation, whereas ARID2 mutation often occurred later. Transcriptome analyses revealed the epithelial–mesenchymal transition (EMT) and hypoxic phenotype in sarcomatoid tumor components, which were confirmed by immunohistochemical staining. Moreover, we identified ARID2 mutations in 70% (7/10) of patients with sarcomatoid HCC but only 1–5% of patients with non-sarcomatoid HCC. Biofunctional investigations revealed that inactivating mutation of ARID2 contributes to HCC growth and metastasis and induces EMT in a hypoxic microenvironment. Conclusions We offer a comprehensive description of the molecular basis for sarcomatoid HCC, and identify genomic alteration (ARID2 mutation) together with the tumor microenvironment (hypoxic microenvironment), that may contribute to the formation of the sarcomatoid tumor component through EMT, leading to sarcomatoid HCC development and progression.

OBJECTIVE: To investigate whether Buyang Huanwu Decoction (BYHWD) had a good curative effect on the neuroprotection of red nucleus neurons after spinal cord injury (SCI) and the possible molecular mechanism. METHODS: Ninety male Sprague-Dawley rats were divided into 5 groups (n=18 per group) according to a random number table, including the control, model, low- (12.78 g/kg, BL group), medium- (25.65 g/kg, BM group), and high-dose BYHWD groups (51.30 g/kg, BH group). A rubrospinal tract transection model in rats was established, and different doses of BYHWD were intragastrically administrated for 4 weeks. The forelimb locomotor function was recorded using the spontaneous vertical exploration test. Cyclic adenosine monophosphate (cAMP) level in red nucleus was detected through an enzyme-linked immunosorbent assay. The morphology and number of red nucleus neurons were observed using Nissl's staining and axonal retrograde tracing by Fluoro-Gold (FG). The expression of cAMP-dependent protein kinase A (PKA), nuclear factor kappa-B (NF-κB) p65, and brain-derived neurotrophic factor (BDNF) in red nucleus were detected using immunohistochemistry and quantitative real-time polymerase chain reaction. RESULTS: Compared with the control group, the utilization rate of bilateral forelimbs, unilateral right forelimbs, proportion of FG-labeled positive neurons, cAMP level, protein expressions of PKA and BDNF, and BDNF mRNA expression were significantly decreased in the model group (P<0.01), while NF-κB p65 was increased in the model group (P<0.01). Compared with the model group, the utilization rate of bilateral forelimbs and unilateral right forelimbs were significantly higher in the BL, BM and BH groups (P<0.01), the proportion of FG-labeled positive neurons, cAMP level, protein expressions of PKA and BDNF and BDNF mRNA expression in all BYHWD groups were increased (P<0.05 or P<0.01), while NF-κB p65 were decreased in all BYHWD groups (P<0.05 or P<0.01). CONCLUSIONS BYHWD possesses a sound neuroprotective effect on red nucleus neurons after SCI, and the efficacy was dose-related. The mechanism may be related to regulating the cAMP/PKA/NF-κ B p65 signaling pathway, finally promoting expression of BDNF.
Animals ; Spinal Cord Injuries/pathology* ; Drugs, Chinese Herbal/therapeutic use* ; Rats, Sprague-Dawley ; Male ; Cyclic AMP/metabolism* ; Transcription Factor RelA/metabolism* ; Cyclic AMP-Dependent Protein Kinases/metabolism* ; Signal Transduction/drug effects* ; Brain-Derived Neurotrophic Factor/genetics* ; Red Nucleus/metabolism* ; Recovery of Function/drug effects* ; Neurons/metabolism* ; Rats

OBJECTIVE: To explore the association between acupuncture during controlled ovarian hyperstimulation (COH) and the live birth rate (LBR) using different propensity score methods. METHODS: In this retrospective cohort study, eligible women who underwent a COH were divided into acupuncture and non-acupuncture groups. The primary outcome was LBR, as determined by propensity score matching (PSM). LBR was defined as the delivery of one or more living infants that reached a gestational age over 28 weeks after embryo transfer. The propensity score model encompassed 16 confounding variables. To validate the results, sensitivity analyses were conducted using three additional propensity score methods: propensity score adjustment, inverse probability weighting (IPW), and IPW with a "doubly robust" estimator. RESULTS: The primary cohort encompassed 9751 patients (1830 [18.76%] in the acupuncture group and 7921 [81.23%] in the non-acupuncture group). Following 1:1 PSM, a higher LBR was found in the acupuncture cohort (41.4% [755/1824] vs 36.4% [664/1824], with an odds ratio of 1.23 [95% confidence interval, 1.08-1.41]). Three additional propensity score methods produced essentially similar results. The risk of serious adverse events did not significantly differ between the two groups. CONCLUSION This retrospective study revealed an association between acupuncture and an increased LBR among patients undergoing COH, and that acupuncture is a safe and valuable treatment option. Please cite this article as: Zheng XY, Jiang ZY, Li YT, Li CL, Zhu H, Yu Z, Yu SY, Yang LL, Tang SY, Lü XY, Liang FR, Yang J. Association between acupuncture and live birth rates after fresh embryo transfer: A cohort study based on different propensity score methods. J Integr Med. 2025; 23(5):528-536.
Humans ; Female ; Propensity Score ; Embryo Transfer ; Adult ; Acupuncture Therapy ; Retrospective Studies ; Pregnancy ; Live Birth ; Birth Rate ; Cohort Studies

Medical institutions, with their clinical practice foundation and abundant human use experience data, have become important carriers for the inheritance and innovation of traditional Chinese medicine(TCM) and the "cradles" of the preparation of new TCM. To effectively promote the transformation of new TCM originating from the TCM clinical practice in medical institutions and establish an effective evaluation index system for the transformation of new TCM conforming to the characteristics of TCM, consensus experts adopted the literature research, questionnaire survey, Delphi method, etc. By focusing on the policy and technical evaluation of new TCM originating from the TCM clinical practice in medical institutions, a comprehensive evaluation from the dimensions of drug safety, efficacy, feasibility, and characteristic advantages was conducted, thus forming a comprehensive evaluation system with four primary indicators and 37 secondary indicators. The expert consensus reached aims to encourage medical institutions at all levels to continuously improve the high-quality research and development and transformation of new TCM originating from the TCM clinical practice in medical institutions and targeted at clinical needs, so as to provide a decision-making basis for the preparation, selection, cultivation, and transformation of new TCM for medical institutions, improve the development efficiency of new TCM, and precisely respond to the public medication needs.
Medicine, Chinese Traditional/standards* ; Humans ; Consensus ; Drugs, Chinese Herbal/therapeutic use* ; Surveys and Questionnaires