AIM:To explore the influence of microRNA-193b-5p(miR-193b-5p)on apoptosis of cardiomyo-cytes in an ischemia-hypoxia(IH)environment and the possible mechanism.METHODS:Human AC16 cardiomyocytes were cultured in vitro,and an IH model of cardiomyocytes was established.The cardiomyocytes were divided into control group,IH group,IH+miR-193b-5p inhibitor group,and IH+inhibitor NC group.The cells in control group were regularly cultured,those in IH group were treated with a hypoxia chamber for 24 h to induce cardiomyocyte apoptosis,while those IH+miR-193b-5p inhibitor and IH+inhibitor NC groups were transfected with corresponding plasmids by the same opera-tion method and then underwent IH treatment for 24 h to induce cardiomyocyte apoptosis.RT-qPCR was used to detect the expression of miR-193b-5p in cardiomyocytes after IH.The viability of cardiomyocytes was detected by CCK-8 method.Lactate dehydrogenase(LDH)was detected to understand the damage of cardiomyocytes.The apoptotic rate was detected by flow cytometry.The protein levels of Bax,Bcl-2 and cleaved caspase-3 were detected by Western blot.Finally,down-stream target genes were predicted by RNA sequencing combined with bioinformatics methods,and the interaction relation-ship between miR-193b-5p and RING(really interesting new gene)finger protein 4(RNF4)gene was verified by RT-qPCR and Western blot experiments.RESULTS:Compared with control group,miR-193b-5p was highly expressed in cardio-myocytes with IH.Furthermore,in the IH environment,the viability of cardiomyocytes decreased,and cell damage and cell apoptosis increased,while inhibiting the expression level of miR-193b-5p could enhance the viability of cardiomyo-cytes,reduce cell damage,and alleviate the apoptosis of cardiomyocytes induced by IH.The results of RNA sequencing and further experiments verified that miR-193b-5p might act on the RNF4 gene to exert its effect.CONCLUSION:Inhi-bition of miR-193b-5p can alleviate the IH injury and apoptosis of cardiomyocytes.The mechanism might be that miR-193b-5p inhibition exerts a cardioprotective effect against apoptosis by mediating the expression of the RNF4 gene.

Objective:To evaluate the clinical equivalence between a domestic calcipotriol/betamethasone dipropionate ointment and the originator product in the treatment of stable plaque psoriasis.Methods:A multicenter, randomized, double-blind, three-arm, parallel-group, active- and placebo-controlled study was conducted, and 449 patients aged 18 - 65 years with stable plaque psoriasis were enrolled from 25 hospitals (such as the First Affiliated Hospital of China Medical University). Eligible patients had a baseline physician's global assessment (PGA) score of ≥ 3 points, baseline body surface area (BSA) involvement of 5% - 30%, and a target lesion psoriasis area and severity index (TL-PASI) for plaque elevation of ≥ 3 points. Participants were randomly assigned in a 2:2:1 ratio to the test group ( n = 179), reference group ( n = 180), and placebo group ( n = 90), and applied the domestic calcipotriol/betamethasone dipropionate ointment, originator product, and ointment base respectively, once daily in the evening for 4 weeks. Efficacy and safety were assessed at weeks 1, 2, and 4. The primary efficacy endpoints were the treatment success rates and clinical success rates in each group at week 4. The per-protocol set (PPS) was used for the primary efficacy analysis, and the intention-to-treat (ITT) set for supplementary efficacy analysis. Equivalence between the test and reference preparations was tested using the Cochran-Mantel-Haenszel method adjusted for randomization strata. Superiority of the test and reference preparations over the placebo was also tested. Measurement data were compared among the 3 groups using analysis of variance or non-parametric tests, while treatment success rates, clinical success rates, and incidence rates of adverse reactions were compared using the chi-square test. Results:The ITT, PPS, and safety sets included 447, 420, and 448 patients, respectively. In the ITT set, patients were aged 43.6 ± 12.8 years, including 320 (71.6%) males and 127 (28.4%) females, and the disease duration was 11.21 ± 9.05 years; 316 (70.7%) had a PGA score of 3 points and 131 (29.3%) had a PGA score of 4 - 5 points. No significant differences in the baseline characteristics (including age, sex, disease duration and disease severity) were observed among the 3 groups (all P > 0.05). Based on the PPS analysis, the treatment success rates were 57.9% (99/171) in the test group, 50.3% (86/171) in the reference group, and 7.7% (6/78) in the placebo group, and the clinical success rates were 57.9% (99/171), 50.3% (86/171), and 10.3% (8/78), respectively; both the test and reference groups were superior to the placebo group in both treatment and clinical success rates (all P < 0.001) ; the rate differences for treatment success (90% confidence interval [ CI]: -1.3% - 16.4%) and clinical success (90% CI: -1.3% - 16.3%) between the test and reference groups were entirely within the pre-defined equivalence margin (-20% - 20%). Subgroup analyses by baseline PGA scores: for patients with a baseline PGA score of 3 points, the treatment success rates in the test, reference, and placebo groups were 60.8% (73/120), 52.1% (62/119), and 11.1% (6/54), respectively, and the corresponding clinical success rates were 61.7% (74/120), 53.8% (64/119), and 13% (7/54), respectively; the test and reference groups did not differ significantly in treatment or clinical success rates (both P > 0.05), but both showed higher success rates than the placebo group (all P < 0.001) ; the results of statistical comparisons among the 3 groups in patients with a baseline PGA score of 4 - 5 points were consistent with those observed in patients with a baseline PGA score of 3 points. The percentage reductions in PGA and TL-PASI scores from baseline to weeks 1, 2, and 4 showed significant differences among the 3 groups, which were significantly higher in the test and reference groups than in the placebo group (all P < 0.001), but did not differ between the test and reference groups (all P > 0.05). The primary adverse reactions were local skin reactions, such as pruritus, pain, and erythema. The incidence rates of adverse reactions were 8.9% (16/179) in the test group, 7.3% (13/179) in the reference group, and 7.8% (7/90) in the placebo group, with no significant difference among the 3 groups ( P > 0.05) . Conclusions:The domestic calcipotriol/betamethasone dipropionate ointment demonstrated clinical equivalence to the originator product in the treatment of stable plaque psoriasis, and the two agents exhibited comparable efficacy for patients with varying degrees of disease severity, and were comparable in the speed and degree of clinical improvement, with similar favorable safety profiles.

Objective:To evaluate the clinical equivalence between a domestic calcipotriol/betamethasone dipropionate ointment and the originator product in the treatment of stable plaque psoriasis.Methods:A multicenter, randomized, double-blind, three-arm, parallel-group, active- and placebo-controlled study was conducted, and 449 patients aged 18 - 65 years with stable plaque psoriasis were enrolled from 25 hospitals (such as the First Affiliated Hospital of China Medical University). Eligible patients had a baseline physician's global assessment (PGA) score of ≥ 3 points, baseline body surface area (BSA) involvement of 5% - 30%, and a target lesion psoriasis area and severity index (TL-PASI) for plaque elevation of ≥ 3 points. Participants were randomly assigned in a 2:2:1 ratio to the test group ( n = 179), reference group ( n = 180), and placebo group ( n = 90), and applied the domestic calcipotriol/betamethasone dipropionate ointment, originator product, and ointment base respectively, once daily in the evening for 4 weeks. Efficacy and safety were assessed at weeks 1, 2, and 4. The primary efficacy endpoints were the treatment success rates and clinical success rates in each group at week 4. The per-protocol set (PPS) was used for the primary efficacy analysis, and the intention-to-treat (ITT) set for supplementary efficacy analysis. Equivalence between the test and reference preparations was tested using the Cochran-Mantel-Haenszel method adjusted for randomization strata. Superiority of the test and reference preparations over the placebo was also tested. Measurement data were compared among the 3 groups using analysis of variance or non-parametric tests, while treatment success rates, clinical success rates, and incidence rates of adverse reactions were compared using the chi-square test. Results:The ITT, PPS, and safety sets included 447, 420, and 448 patients, respectively. In the ITT set, patients were aged 43.6 ± 12.8 years, including 320 (71.6%) males and 127 (28.4%) females, and the disease duration was 11.21 ± 9.05 years; 316 (70.7%) had a PGA score of 3 points and 131 (29.3%) had a PGA score of 4 - 5 points. No significant differences in the baseline characteristics (including age, sex, disease duration and disease severity) were observed among the 3 groups (all P > 0.05). Based on the PPS analysis, the treatment success rates were 57.9% (99/171) in the test group, 50.3% (86/171) in the reference group, and 7.7% (6/78) in the placebo group, and the clinical success rates were 57.9% (99/171), 50.3% (86/171), and 10.3% (8/78), respectively; both the test and reference groups were superior to the placebo group in both treatment and clinical success rates (all P < 0.001) ; the rate differences for treatment success (90% confidence interval [ CI]: -1.3% - 16.4%) and clinical success (90% CI: -1.3% - 16.3%) between the test and reference groups were entirely within the pre-defined equivalence margin (-20% - 20%). Subgroup analyses by baseline PGA scores: for patients with a baseline PGA score of 3 points, the treatment success rates in the test, reference, and placebo groups were 60.8% (73/120), 52.1% (62/119), and 11.1% (6/54), respectively, and the corresponding clinical success rates were 61.7% (74/120), 53.8% (64/119), and 13% (7/54), respectively; the test and reference groups did not differ significantly in treatment or clinical success rates (both P > 0.05), but both showed higher success rates than the placebo group (all P < 0.001) ; the results of statistical comparisons among the 3 groups in patients with a baseline PGA score of 4 - 5 points were consistent with those observed in patients with a baseline PGA score of 3 points. The percentage reductions in PGA and TL-PASI scores from baseline to weeks 1, 2, and 4 showed significant differences among the 3 groups, which were significantly higher in the test and reference groups than in the placebo group (all P < 0.001), but did not differ between the test and reference groups (all P > 0.05). The primary adverse reactions were local skin reactions, such as pruritus, pain, and erythema. The incidence rates of adverse reactions were 8.9% (16/179) in the test group, 7.3% (13/179) in the reference group, and 7.8% (7/90) in the placebo group, with no significant difference among the 3 groups ( P > 0.05) . Conclusions:The domestic calcipotriol/betamethasone dipropionate ointment demonstrated clinical equivalence to the originator product in the treatment of stable plaque psoriasis, and the two agents exhibited comparable efficacy for patients with varying degrees of disease severity, and were comparable in the speed and degree of clinical improvement, with similar favorable safety profiles.

AIM:To explore the influence of microRNA-193b-5p(miR-193b-5p)on apoptosis of cardiomyo-cytes in an ischemia-hypoxia(IH)environment and the possible mechanism.METHODS:Human AC16 cardiomyocytes were cultured in vitro,and an IH model of cardiomyocytes was established.The cardiomyocytes were divided into control group,IH group,IH+miR-193b-5p inhibitor group,and IH+inhibitor NC group.The cells in control group were regularly cultured,those in IH group were treated with a hypoxia chamber for 24 h to induce cardiomyocyte apoptosis,while those IH+miR-193b-5p inhibitor and IH+inhibitor NC groups were transfected with corresponding plasmids by the same opera-tion method and then underwent IH treatment for 24 h to induce cardiomyocyte apoptosis.RT-qPCR was used to detect the expression of miR-193b-5p in cardiomyocytes after IH.The viability of cardiomyocytes was detected by CCK-8 method.Lactate dehydrogenase(LDH)was detected to understand the damage of cardiomyocytes.The apoptotic rate was detected by flow cytometry.The protein levels of Bax,Bcl-2 and cleaved caspase-3 were detected by Western blot.Finally,down-stream target genes were predicted by RNA sequencing combined with bioinformatics methods,and the interaction relation-ship between miR-193b-5p and RING(really interesting new gene)finger protein 4(RNF4)gene was verified by RT-qPCR and Western blot experiments.RESULTS:Compared with control group,miR-193b-5p was highly expressed in cardio-myocytes with IH.Furthermore,in the IH environment,the viability of cardiomyocytes decreased,and cell damage and cell apoptosis increased,while inhibiting the expression level of miR-193b-5p could enhance the viability of cardiomyo-cytes,reduce cell damage,and alleviate the apoptosis of cardiomyocytes induced by IH.The results of RNA sequencing and further experiments verified that miR-193b-5p might act on the RNF4 gene to exert its effect.CONCLUSION:Inhi-bition of miR-193b-5p can alleviate the IH injury and apoptosis of cardiomyocytes.The mechanism might be that miR-193b-5p inhibition exerts a cardioprotective effect against apoptosis by mediating the expression of the RNF4 gene.

Objective To explore the effect of intermittent fasting combined with diversified management mode on weight loss of overweight and obese people. Methods A total of 120 overweight and obese patients were selected as research objects, and randomly divided into control group (n=60) and experimental group (n=60). The control group was given intermittent fasting, and the experimental group was given diversified management mode on the basis of the control group. Waist circumference, body mass, body mass index (BMI), waist-to-hip ratio and blood lipid level were compared between the two groups; the scores of hunger, satiety, satisfaction and the incidence of adverse reactions were compared between the two groups. Results After intervention, waist circumference, body mass, BMI, waist-to-hip ratio, total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in the experimental group were significantly lower, and high-density lipoprotein cholesterol (HDL-C) was significantly higher than that in the control group (P < 0.05). After intervention, the hunger score of the experimental group was significantly lower, and the satiety score and satisfaction score were significantly higher than those of the control group (P < 0.05). The total incidence of adverse reactions in the experimental group was 6.67%, which was significantly lower than 20.00% in the control group (P < 0.05). Conclusion The application of intermittent fasting combined with diversified management mode in overweight and obese people is better, which can improve the relevant clinical indicators and blood lipid levels, and has high safety.

Objective: To investigate the characteristics of fall injuries among primary and middle school students in Yinzhou District, Ningbo City, Zhejiang Province from 2010 to 2022, so as to provide insights into prevention and control of fall injuries among primary and middle school students. Methods: Data of 6 to 17 year-old primary and middle school students with initial diagnosis of fall injury from 2010 to 2022 were collected through Injury Subsystem of Zhejiang Provincial Chronic Disease Surveillance Information Management System. The time, place, activity and clinical characteristics of fall injury were analyzed using a descriptive method. Results: Totally 7 808 fall injury cases among primary and middle school students were reported in Yinzhou District from 2010 to 2022, accounting for 45.72% of the total injury cases in the same age. There were 5 413 boys and 2 395 girls, with a boy/girl ratio of 2.26∶1. Primary school students were the majority, accounting for 55.10%. The incidence of fall injuries among girls, junior high school students and high school students showed increasing trends from 2012 to 2022 (both P<0.05). The primary peak of fall injury was from September to November, and the secondary peak was from May to July, with 4 510 cases (57.76%). The place of fall injury development mainly included schools (2 680 cases, 34.32%), homes (2 343 cases, 30.01%) and streets/urban areas (2 247 cases, 28.78%). The activities at the time of fall injury mainly included leisure time (3 490 cases, 44.70%), sports (2 861 cases, 36.64%) and school activities (1 094 cases, 14.01%). Soft tissue injury was the main characteristics (6 224 cases, 79.71%). Lower limbs (3 101 cases, 39.72%), head (2 419 cases, 30.98%) and upper limbs (1 974 cases, 25.28%) were the main injury sites. Mild injury was predominant (5 896 cases, 75.51%). Conclusions Boys and primary school students are high-risk groups of fall injury in Yinzhou District, schools are high-risk places of fall injury. Schools should be regarded as key intervention places and health education on fall injury prevention should be strengthened.

Objective: To analyze the association between latent class of health-risk behaviors and depressive symptoms among middle school students, so as to provide the evidence for the prevention and intervention of depressive symptoms among middle school students. Methods: Students in two junior high schools, two senior high schools and one vocational high school in Yinzhou District, Ningbo City, Zhejiang Province, were selected using a stratified multi-stage cluster sampling method. Demography and health-risk behaviors were collected using questionnaire surveys, depressive symptoms were investigated using the Center for Epidemiological Studies Depression-10 Scale, and latent class analysis was conducted for health-risk behaviors. The association between different latent classes and depressive symptoms was analyzed using a multivariable logistic regression model. Results: A total of 1 247 students were surveyed, including 641 boys (51.40%) and 606 girls (48.60%). There were 452 junior high school students (36.25%), 532 high school students (42.66%) and 263 vocational high school students (21.09%). Latent class analysis showed that health-risk behaviors in students were classified into three groups, namely healthy behavior group (52.93%), poor diet group (39.94%) and high-risk behavior group (7.14%), and the detection rates of depressive symptoms were 7.12%, 18.88% and 52.81%, respectively, with a statistically significant difference between groups (P<0.05). Multivariable logistic regression analysis showed that after adjusting for age, gender, native place, only child and living on campus, the poor diet group (OR=3.107, 95%CI: 2.086-4.627) and high-risk behavior group (OR=15.401, 95%CI: 9.031-26.262) had higher risks of depressive symptoms compared with the healthy behavior group. Conclusion Having high-risk behaviors and poor diet may increase the risk of developing depressive symptoms among middle school students.

Objective To analyze the deep venous thrombosis(DVT)after plasma infusion in a patient with congenital dysfibrinogene-mia(CD),and explore the relationship between the CD and DVT.Methods The clinical data were collected and the pedigree was investigated(3 subjects of 2 generations in total).The relevant indexes of coagulation factors of the patient and her family members were detected.The genomic DNA of peripheral blood was extracted for PCR amplification.All the exons,flanking sequences,5'and 3'untranslated regions of FGA,FGB and FGG genes of fibrinogen(Fg)of the patient were analyzed by direct sequencing.The corre-sponding mutation site was subjected to sequence in the other members of this family.The PyMol software was used to construct the pro-tein model before and after gene mutation.Results The patient was admitted to hospital for hysteromyomectomy.DVT appeared in 3 days after surgery.The prothrombin time(PT),thrombin time(TT),Fg activity(Fg∶C)and Fg antigen(Fg∶Ag)of the patient was 14.9 s,33.3 s,0.94 g/L and 2.10 g/L,respectively.The above four indicators in her mother were 14.7 s,32.8 s,0.97 g/L and 2.35 g/L,respectively.Gene sequencing revealed that both the patient and her mother had a heterozygous missense mutation c.2185G＞A(p.Glu729Lys)in exon 6 of the FGA gene.The protein model analysis demonstrated that p.Glu729Lys mutation changed the amino acid side chain and reduced the number of hydrogen bonds originally formed with Arg854.Conclusion A heterozygous missense mutation c.2185G＞A(NM_000508)in exon 6 of the FGA gene should be responsible for the low fibrinogen level in this pedigree,which might be the main reason for DVT after plasma infusion in this patient.

Objective:To investigate the role of m.4435A>G and YARS2 c.572G>T(p.G191V)mutations in the development of essential hypertension.Methods:A hypertensive patient with m.4435A>G and YARS2 p.G191V mutations was identified from previously collected mitochondrial genome and exon sequencing data.Clinical data were collected,and a molecular genetic study was conducted in the proband and his family members.Peripheral venous blood was collected,and immortalized lymphocyte lines constructed.The mitochondrial transfer RNA(tRNA),mitochondrial protein,adenosine triphosphate(ATP),mitochondrial membrane potential(MMP),and reactive oxygen species(ROS)in the constructed lymphocyte cell lines were measured.Results:Mitochondrial genome sequencing showed that all maternal members carried a highly conserved m.4435A>G mutation.The m.4435A>G mutation might affect the secondary structure and folding free energy of mitochondrial tRNA and change its stability,which may influence the anticodon ring structure.Compared with the control group,the cell lines carrying m.4435A>G and YARS2 p.G191V mutations had decreased mitochondrial tRNA homeostasis,mitochondrial protein expression,ATP production and MMP levels,as well as increased ROS levels(all P<0.05).Conclusion:The YARS2 p.G191V mutation aggravates the changes in mitochondrial translation and mitochondrial function caused by m.4435A>G through affecting the steady-state level of mitochondrial tRNA and further leads to cell dysfunction,indicating that YARS2 p.G191V and m.4435A>G mutations have a synergistic effect in this family and jointly participate in the occurrence and development of essential hypertension.

OBJECTIVE：To establish the UPLC fingerprint of Polygonum cuspidatum ，and to determine the contents of four active ingredients and to provide reference for the quality evaluation of P. cuspidatum . METHODS ：The determination was performed on Waters BEH C 18 column（100 mm×2.1 mm，1.7 μm）with mobile phase consisted of acetonitrile- 0.2％ formic acid （gradient elution ）at flow rate of 0.4 mL/min. The column temperature was 40 ℃，and detection wavelength was 290 nm. The sample size was 1 μL. The fingerprints were evaluated by similarity calculation，cluster analysis and orthogonal partial least square discriminant analysis （OPLS-DA）. Using polydatin as internal standard ，relative calibration factors of resveratrol ，emodin-8-O- β-D-glucoside and emodin were determined to develop a method of QAMS. The contents of 4 above components in 15 batches of P. cuspidatum were calculated by relative calibration factors. The results of QAMS were compared with those of external standard. RESULTS：UPLC fingerprints of 15 batches of P. cuspidatum were established ，and 12 common peaks were confirmed. Five components were identified ，i.e. polydatin ，resveratrol，emodin-8-O-β-D-glucoside，emodin，emodin methyl ether. The fingerprint similarity of 15 batches of P. cuspidatum was in the range of 0.865-0.976. According to cluster analysis ，15 batches of P. cuspidatum were classified into 4 categories，showing certain regularity of origin. Seven markers were identified by OPLS-DA method. The order of difference significance was peak 7＞emodin-8-O-β-D-glucoside＞resveratrol＞peak 8＞polydatin＞peak 1＞ peak 10. The relative deviation among the contents of resveratrol ，emodin-8-O-β-D-glucoside and emodin in 15 batches of P. cuspidatum determined by QAMS and external standard method was less than 5.0％，indicating that there was no significant difference between the two methods. CONCLUSIONS ：UPLC fingerprint combined with QAMS method is convenient and reliable for the quality evaluation of P. cuspidatum ；the quality of P. cuspidatum produced in Chongqing and Anhui province is better.