Objective To explore method for improving the colonization efficiency of Helicobacter pylori(Hp)in the mouse stomach and to determine if the proton pump inhibitor(PPI)pantoprazole can act as a colonization adjuvant to enhance Hp colonization,with the aim of providing an effective tool for establishing an Hp infection mouse model.Methods The Hp Sydney strain 1(SS1)was introduced and solid plate and liquid culture systems were established.The effects of different doses of pantoprazole on gastric acid secretion in mice were compared.The impact of Hp inoculation,alone or combined with pantoprazole pretreatment,on Hp colonization efficiency was analyzed using rapid urease tests,bacterial plate cultures,and TaqMan quantitative polymerase chain reaction.Results PPI pretreatment inhibited gastric acid secretion and promoted Hp colonization in the mouse stomach,to some extent.Conclusions PPI can serve as colonization adjuvants to enhanc e the efficiency of constructing Hp infection mouse models.

Objective:To investigate the clinical efficacy of cervical cerclage in singleton pregnancy with different degrees of short cervix.Methods:The clinical data of singleton pregnant women who underwent transvaginal ultrasound examination at 18-24 +6 weeks of gestation and found cervical dilation with cervical length (CL) ≤20 mm, and without history of spontaneous preterm delivery or late abortion in Women′s Hospital, Zhejiang University School of Medicine from January 2021 to September 2023 were collected and retrospectively analyzed. According to the case control matching, 78 pregnant women in the cerclage group and 78 women in the conservative treatment group were finally included. The pregnancy outcomes and neonatal prognosis of the two groups were compared. Meanwhile, the two groups of pregnant women were divided into three subgroups for stratification (CL≤10, 11-15, 16-20 mm). Multivariate logistic regression analysis and Kaplan-Meier curve were used to evaluate the effect of cervical cerclage on pregnancy outcomes in different subgroups. Results:(1) Compared with the conservative treatment group, the gestational age at delivery (median: 36 vs 37 weeks) and the duration of pregnancy extension (median: 90 vs 97 days) in the cerclage group were not significantly prolonged (all P>0.05). There was no significant difference in the cumulative non-delivery rate between the cerclage group and the conservative treatment group ( P=0.143). The rate of neonatal intensive care unit (NICU) admission in the cerclage group was significantly higher than that in the conservative treatment group (46.1% vs 31.2%, P<0.001), but there were no significant differences in other neonatal outcomes between the two groups (all P>0.05). (2) CL 16-20 mm subgroup: compared with the conservative treatment group (28 cases), the cerclage group (18 cases) had a significantly higher rate of preterm birth before 37 weeks of gestation (3.6% vs 6/18, P<0.001) and a significantly lower neonatal birth weight (median: 3 370 vs 2 925 g, P<0.001). There was no significant difference in the cumulative non-delivery rate between the cerclage group and the conservative treatment group ( P=0.168). (3) CL 11-15 mm subgroup: compared with the conservative treatment group (26 cases), the gestational age of delivery in the cerclage group (32 cases) was later (median: 36 and 37 weeks, respectively), and the difference was statistically significant ( P<0.05). The cumulative non-delivery rate in the cerclage group was significantly higher than that in the conservative treatment group ( P=0.001). (4) CL≤10 mm subgroup: compared with the conservative treatment group (24 cases), the pregnant women in the cerclage group (28 cases) had a later gestational age at delivery (median: 34 vs 37 weeks), a longer duration of pregnancy (median: 74 vs 97 days), and a larger newborn birth weight (median: 2 300 vs 3 165 g). The rates of preterm birth before 34 weeks of gestation (45.8% vs 14.3%) and before 37 weeks of gestation (83.3% vs 39.3%) were lower, and the differences were all statistically significant (all P<0.05). The cumulative non-delivery rate of pregnant women in the cerclage group was significantly higher than that in the conservative treatment group ( P<0.001). Conclusion:Cervical cerclage could significantly prolong the gestational weeks and improve the perinatal outcomes for singleton pregnant women with cervical internal orifice dilation and CL≤15 mm without a history of spontaneous preterm delivery or late abortion.

Objective To explore the roles of the mechanoreceptor Yes-associated protein 1(YAP1)and piezo type mechanosensitive ion channel component 2(PIEZO2)in mechanotransduction in mouse bladder urothelial cells.Methods Mouse bladder urothelial cells were subjected to mechanical stretching using the FX-6000T cell stretching system and treated with the YAP1-specific inhibitor verteporfin(VP).The expressions of PIEZO2,YAP1 and connective tissue growth factor(CTGF)at the mRNA and protein levels,as well as changes in cellular adenosine triphosphatase(ATP)concentration,were detected using reverse transcription quantitative PCR(RT-qPCR)and Western blotting(WB).Results After stretching stimulation,under the fluorescence microscope,it was observed that the diameter length of the stretched cells were longer than that before stretching,and the difference was statistically significant(P＜0.05).The expressions of YAP1,PIEZO2 and CTGF at the mRNA and protein levels were increased in the stretched group compared to those of the non-stretched group(P＜0.05).VP effectively reduced the expressions of YAP1,PIEZO2 and CTGF at the mRNA and protein levels after stretching stimulation(P＜0.05).Stretching stimulation significantly increased the intracellular ATP concentration,while VP was able to inhibit the increase in ATP concentration,with a statistically significant difference(P＜0.000 1).Conclusion Stretching stimulation increased the expressions of YAP1 and PIEZO2 in bladder urothelial cells and promoted the release of ATP;verteporfin inhibited the increase in YAP1 activity and the overexpression of PIEZO2 caused by stretching,thereby reducing the release of ATP.It is suggested that mouse bladder urothelial cells may primarily sense mechanical signals through the YAP1-PIEZO2-ATP pathway.

Objective:To examine the clinical effects of the SCERTS model-based peer-mediated intervention (PMI) on the social skills of children with autism spectrum disorder (ASD) and provide new approaches to the rehabilitation treatment of ASD children.Methods:A randomized controlled study.A total of 120 children with mild-to-moderate ASD diagnosed Xi′an Traditional Chinese Medicine Hospital of Encephalopathy between April 2023 and April 2024 were selected.They were assigned to either the experimental or control group using the random number table method, with 60 cases in each group.The experimental group was treated with the SCERTS model-based PMI, whereas the control group underwent conventional rehabilitation training, comprising cognitive, language and behavioural interventions, acupuncture and other techniques.Changes in Social Responsiveness Scale (SRS), Autism Social Skills Scale (ASSS) and Autism Treatment Evaluation Checklist (ATEC) scores were observed in the 2 groups of children before and after treatment.The statistical analyses were performed using a paired-samples t-test for intra-group comparisons and an independent-samples t-test for inter-group comparisons. Results:Following 12 weeks of intervention, the SRS and ATEC scores decreased while the ASSS scores increased after treatment in both groups.The differences were statistically significant (all P<0.05).The SRS [(83.25±14.56) points] and ATEC [(79.41±15.36) points] of the experimental group were lower than those of the control group [SRS(89.80±12.69) points, ATEC(85.95±16.13) points].The ASSS score of the experimental group was higher than that of the control group [(112.77±22.42) points vs.(103.80±24.13) points] ( t=2.627, 2.274, -2.109; all P<0.05). Conclusions:The SCERTS model-based PMI is an efficacious approach for the improvement of social skills in ASD children, and thus merits further investigation and application.

Objective To explore the role of secreted phosphoprotein 1(SPP1)and myeloid differentiation primary response 88(MYD88)in morphine-induced cardiomyocyte apoptosis.Methods A morphine addiction model was established in Sprague-Dawley(SD)rats.Twelve SD rats were randomly assigned to the normal saline(NS)group or the morphine-dependent(DAM)group.Histopathological analysis was employed to observe and compare myocardial tissue morphology between the two groups.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was performed to assess the number of apoptotic cells in each group.The expression levels of SPP1 and MYD88 were evaluated using immunohistochemistry.Quantitative real-time poly merase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression of SPP1,MYD88,Bax,Bcl2,Caspase-3,and Caspase-9.Simultaneously,Western blot analysis was used to detected the expression of Cleaved Caspase-3 and Cleaved Caspase-9 proteins.In vitro,SPP1 expression was knocked down in primary neonatal rat cardiomyocytes(NRCMs),and cells were divided into three groups:control(CON),morphine treated(DA),and shSPP1#3+DA.Cell viability was assessed using the CCK-8 assay,and apoptosis rates were determined by flow cytometry.Results HE and TUNEL staining of myocardial tissues from morphine-addicted SD rats revealed that,compared with the NS group,myofibrils in the DAM group exhibited partial disruption and a significant increase in apoptotic cells(P<0.05).Western blot and RT-qPCR analyses demonstrated that,relative to the NS group,the mRNA and protein levels of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 were significantly upregulated in the DAM group(P<0.05),whereas Bcl2 expression was significantly downregulated at both mRNA and protein levels(P<0.05),and the protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were also increased.with all differences being statistically significant.In NRCMs following morphine intervention,cell viability in the DA group was markedly reduced compared to the CON group(P<0.05),accompanied by a signifi-cant increase in apoptosis rate(P<0.05).Consistently,Western blot and RT-qPCR results showed elevated mRNA and protein expression of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 in the DA group(P<0.05),along with decreased Bcl2 expression(P<0.05).The protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were elevated simultaneously.In contrast,the shSPP1#3+DA group exhibited opposing trends compared to the DA group,with statistically sig nificant differences(P<0.05).Conclusion SPP1 and MYD88 play critical roles in mediating morphine-induced cardiomyocyte apoptosis,and silencing SPP1 has been shown to significantly reduce the extent of cardiomyocyte apoptosis following morphine exposure.

This paper presents a comprehensive study of Pulsatillae Radix,encompassing its pharmacognostic investigation,chemical constituents,and pharmacological effects.Pulsatillae Radix,first recorded in the Shen Nong's Herbal Classic,is known for its efficacy in cooling blood to stop dysentery,clearing heat,and detoxifying,and has been used to treat diseases such as enteritis caused by parasites,demonstrating potential as an antiviral candidate.Research has revealed that its major chemical constituents include triterpenoid saponins,triterpenic acids,and various other components,which collectively constitute the pharmacological foundation of Pulsatillae Radix.Significant pharmacological activities have been observed in antitumor,anti-inflammatory,antimicrobial,and antiviral aspects,providing a scientific basis for its broad application in the pharmaceutical field.

Objective:To investigate the clinical characteristics of omphalocele, and to assess the risk factors associated with adverse outcomes.Methods:A retrospective cohort study was conducted. Clinical data of 224 patients diagnosed with omphalocele, who were hospitalized at Children′s Hospital, Zhejiang University School of Medicine from January 2013 to December 2022, were collected. Based on their discharge outcomes, the patients were classified into 2 groups: favorable outcomes and unfavorable outcomes. Chi-square test or continuity correction χ2 test or Fisher exact probability method, and Mann-Whitney U test were used for intergroup comparisons. Logistic regression analysis was performed to identify risk factors associated with adverse outcomes in omphalocele. Results:Among the 224 patients with omphalocele, 126 were male. A total of 208 patients (92.9%) had favorable outcomes, while 16 patients (7.1%) had unfavorable outcomes. In the unfavorable outcomes group, 14 patients had giant omphaloceles, while 100 patients had giant omphaloceles in the favorable outcomes group. The rates of herniation of more than two intra-abdominal organs in the hernial sac, congenital heart defects, patent ductus arteriosus, pulmonary hypertension, sepsis and infection of the hernial sac, were all higher in the unfavorable outcomes group compared to the favorable outcomes group (all P<0.05). Patients with unfavorable outcomes had longer mechanical ventilation time, duration of oxygen use, duration of parenteral nutrition, hospital stays, and higher rates of parenteral nutrition-associated cholestasis compared to those with favorable outcomes (all P<0.01). Multivariate Logistic regression analysis indicated that pulmonary hypertension ( OR=9.39, 95% CI 1.20-73.32), sepsis ( OR=8.59, 95% CI 1.32-55.86), and congenital heart defects ( OR=6.55, 95% CI 1.11-38.73) were all independent risk factors for adverse outcomes in omphalocele (all P<0.05). Conclusions:Infants with omphalocele are prone to complications such as cardiovascular malformations, infections, and pulmonary hypertension. Adverse outcomes in omphalocele are associated with pulmonary hypertension, sepsis, and congenital heart defects.

Objective:To explore the clinical effects of free bilateral turbocharged anterolateral thigh flaps in tandem in repairing extensive wounds in the foot and ankle.Methods:The study was a retrospective observational study. From April 2020 to June 2023, 12 patients with extensive wounds in the foot and ankle who met the inclusion criteria were admitted to the Department of Wound Repair Surgery of Suzhou Ruihua Orthopedic Hospital, including 8 males and 4 females, aged 21 to 65 years. The wound area after debridement ranged from 27 cm×14 cm to 37 cm×20 cm. The bilateral perforator flaps pedicled with either oblique or descending branches of the lateral circumflex femoral artery were designed and harvested based on the size and shape of the wounds. The individual flap incision area ranged from 16 cm×9 cm to 34 cm×12 cm. The non-homologous perforator of the flap on the one side was turbocharged by anastomosing it with the gross muscular branch or main vessel of the oblique or descending branch of the lateral circumflex femoral artery from the flap. Subsequently, the proximal end of the oblique or descending branch of the lateral circumflex femoral artery and its accompanying vein from the flap on the one side were connected end-to-end with either the anterior tibial artery and vein, posterior tibial artery and vein, or dorsal foot artery and vein in the recipient area, the distal end of the oblique or descending branch of the lateral circumflex femoral artery and its accompanying vein from the flap on the one side were anastomosed end-to-end with a source vessel originating from flap on the other side. The wounds in the flap donor areas were sutured directly. The number and source of perforators carried by the flaps and the duration of the flap repair surgery were recorded. The survival of the flap, the occurrence of vascular crisis, and the wound healing at both donor and recipient areas were observed after surgery. The flap condition, appearance and function of the affected limb were observed during follow-up. At the last follow-up, the sensory function of the flap was assessed using the British Medical Research Council's sensory rating standard, the foot and ankle function of the affected limb was evaluated according to the American Orthopedic Foot and Ankle Society scoring standard.Results:A total 24 flaps were successfully harvested, carrying 60 perforators, including 34 perforators from the oblique branch of the lateral circumflex femoral artery, 24 perforators from the descending branch of the lateral circumflex femoral artery, one perforator from the transverse branch of the lateral circumflex femoral artery, and one perforator from the direct branch of the femoral artery. The duration of the flap repair surgery ranged from 4.2 to 9.0 hours. The flaps of 12 patients exhibited complete survival after surgery. A total of two flaps of two patients experienced venous crisis after surgery but survived through emergency exploration. One patient encountered undesirable wound healing at the donor area of flap on the one side after surgery, which healed after dressing change, debridement, and suturing. The remaining patients' donor area wounds healed. Two patients displayed impaired wound healing in the recipient area, which improved after dressing change and resection of residual sequestrum, and the wounds in the recipient area of other patients healed successfully. During the follow-up of 4-26 months, the flaps demonstrated favorable color and texture, slight edematous appearance, and partial sensory recovery, as well as good aesthetic and functional restoration of the affected limbs. At the last follow-up, the sensory function of the flap was assessed as grade S2 in 9 cases and grade S3 in 3 cases; the foot and ankle function of the affected limb was evaluated as excellent in two cases, good in 9 cases, and fair in one case.Conclusions:The bilateral turbocharged anterolateral thigh flaps have numerous sources of perforators. By implementing supercharging of non-homologous perforators within the flap, the vascular supply to the flap is turbocharged, thereby mitigating the risk of extensive flap necrosis. The flap is an effective approach for repairing extensive wounds in the foot and ankle, resulting in improved function of the affected limb after repair.

Objective:To explore the efficacy and its related factors of rituximab (RTX) in the treatment of children with frequently relapsing nephrotic syndrome/steroid-dependent nephrotic syndrome (FRNS/SDNS).Methods:It was a single-center retrospective study. The clinical data of FRNS/SDNS children first treated with RTX in the First Affiliated Hospital of Sun Yat-sen University from November 1, 2016 to September 1, 2023 were collected. The number of relapse within 1 year before and after RTX treatment, the time to first relapse after RTX treatment, and the time to B-cell reconstitution were analyzed. At the first treatment, a single dose of RTX was given at 375 mg/m 2, with a maximum dose of 500 mg, once a week, for 1 to 4 doses. The count of CD19 + lymphocytes in the peripheral blood of the children was continuously monitored. If B-cell reconstruction was performed, the decision on whether to proceed to the next course of RTX treatment was made based on clinical manifestations. Kaplan-Meier method was used to analyze relapse-free survival rate after receiving RTX. Cox proportional hazards regression model was used to analyze the related factors of relapse after RTX treatment. Results:A total of 98 FRNS/SDNS children receiving RTX treatment were enrolled, including 75 males (76.5%). The age at onset was 4.0 (1.9, 7.1) years and age of receiving RTX was 11.3 (8.5, 13.5) years. There were 90 children (91.8%) achieving complete remission, while 8 patients (8.2%) did not respond to RTX treatment, and 3 patients (3.1%) progressed to end-stage kidney disease after receiving RTX. The relapse-free survival rates at 6 months and 1 year after RTX treatment were 83.3% (75/90) and 57.9% (22/38), respectively. The frequency of relapse 1 year after RTX treatment decreased compared to 1 year before RTX treatment ( Z=-7.398, P<0.001). Compared with children without relapse during the period of B-cell depletion, relapsed children had a higher number of relapse within one year after RTX treatment ( Z=5.246, P<0.001). The time to first relapse after RTX treatment was 8.3 (4.6, 13.9) months in 51 relapse patients. Compared with children receiving 1 dose of RTX in the first course, those receiving 2 or more doses had a longer time to the first relapse ( Z=2.983, P=0.003). There was no statistically significant difference in time to the first relapse between children who received mycophenolate mofetil therapy after RTX treatment and those who didn't ( P>0.05). The reconstruction time of B cells after the first course of RTX was 6.9 (5.3, 9.0) months. Compared to children receiving one dose of RTX in the first course, those receiving two or more doses had a longer B-cell reconstitution time ( Z=2.739, P=0.006). There was no statistically significant difference in B-cell reconstitution time between children who received mycophenolate mofetil therapy after RTX treatment and those who didn't ( P>0.05). Univariate Cox regression analysis showed that recurrence after calcineurin inhibitor (CNI) treatment before RTX treatment and the number of recurrence in one year before RTX treatment were correlated factors of recurrence after RTX treatment (both P<0.05). Multivariate Cox regression analysis showed that recurrence after CNI treatment before RTX treatment was an independent correlated factor of relapse after RTX therapy ( HR=3.496, 95% CI 1.245-9.818, P=0.018). Infusion reactions occurred in 10 patients (10.2%) and infections were observed in 24 patients (24.5%) during B cell depletion. No serious adverse events occurred. Conclusions:RTX is well tolerated and effective in treating FRNS/SDNS. Recurrence after CNI treatment before RTX treatment may be an independent related factor of relapse after RTX treatment.

Objective:To investigate the clinical efficacy of cervical cerclage in singleton pregnancy with different degrees of short cervix.Methods:The clinical data of singleton pregnant women who underwent transvaginal ultrasound examination at 18-24 +6 weeks of gestation and found cervical dilation with cervical length (CL) ≤20 mm, and without history of spontaneous preterm delivery or late abortion in Women′s Hospital, Zhejiang University School of Medicine from January 2021 to September 2023 were collected and retrospectively analyzed. According to the case control matching, 78 pregnant women in the cerclage group and 78 women in the conservative treatment group were finally included. The pregnancy outcomes and neonatal prognosis of the two groups were compared. Meanwhile, the two groups of pregnant women were divided into three subgroups for stratification (CL≤10, 11-15, 16-20 mm). Multivariate logistic regression analysis and Kaplan-Meier curve were used to evaluate the effect of cervical cerclage on pregnancy outcomes in different subgroups. Results:(1) Compared with the conservative treatment group, the gestational age at delivery (median: 36 vs 37 weeks) and the duration of pregnancy extension (median: 90 vs 97 days) in the cerclage group were not significantly prolonged (all P>0.05). There was no significant difference in the cumulative non-delivery rate between the cerclage group and the conservative treatment group ( P=0.143). The rate of neonatal intensive care unit (NICU) admission in the cerclage group was significantly higher than that in the conservative treatment group (46.1% vs 31.2%, P<0.001), but there were no significant differences in other neonatal outcomes between the two groups (all P>0.05). (2) CL 16-20 mm subgroup: compared with the conservative treatment group (28 cases), the cerclage group (18 cases) had a significantly higher rate of preterm birth before 37 weeks of gestation (3.6% vs 6/18, P<0.001) and a significantly lower neonatal birth weight (median: 3 370 vs 2 925 g, P<0.001). There was no significant difference in the cumulative non-delivery rate between the cerclage group and the conservative treatment group ( P=0.168). (3) CL 11-15 mm subgroup: compared with the conservative treatment group (26 cases), the gestational age of delivery in the cerclage group (32 cases) was later (median: 36 and 37 weeks, respectively), and the difference was statistically significant ( P<0.05). The cumulative non-delivery rate in the cerclage group was significantly higher than that in the conservative treatment group ( P=0.001). (4) CL≤10 mm subgroup: compared with the conservative treatment group (24 cases), the pregnant women in the cerclage group (28 cases) had a later gestational age at delivery (median: 34 vs 37 weeks), a longer duration of pregnancy (median: 74 vs 97 days), and a larger newborn birth weight (median: 2 300 vs 3 165 g). The rates of preterm birth before 34 weeks of gestation (45.8% vs 14.3%) and before 37 weeks of gestation (83.3% vs 39.3%) were lower, and the differences were all statistically significant (all P<0.05). The cumulative non-delivery rate of pregnant women in the cerclage group was significantly higher than that in the conservative treatment group ( P<0.001). Conclusion:Cervical cerclage could significantly prolong the gestational weeks and improve the perinatal outcomes for singleton pregnant women with cervical internal orifice dilation and CL≤15 mm without a history of spontaneous preterm delivery or late abortion.