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 investigate the impact of mitophagy,mediated by the long non-coding RNA SNHG16(LncRNA SNHG16)on diabetes-associated cognitive impairment(DCI).Methods 29 male C57BL/J mice were randomly divided into normal control(NC)group,DCI group and DCI+mitochondrial autophagy inhibitor(DCI+Mdivi-1)group.Morris water maze and new object recognition test were used to detect the cognitive function of mice,qRT-CPR was used to detect the expression of LncRNA SNHG16 and mitochondrial autophagy marker mRNA.Western blot were used to detect the expression of related protein.The mouse hippocampal neurons HT22 were divided into control(Con)group,high glucose(HG)group,HG+SNHG16 silencing(HG+sh-SNHG16)group and HG+no-load control(HG+sh-NC)group.CCK8 method and lactate dehydrogenase(LDH)method were used to detect neuronal damage.JC-1 method was used to detect mitochondrial membrane potential.Results Compared with NC group,the expression of LncRNA SNHG16 and the expression of autophagy-related gene 5,PTEN-induced putative kinase 1(PINK1),Parkin and microtubule associated protein light chain 3(LC3)Ⅱ/Ⅰ increased(P<0.05),while the expression of mitochondrial autophagy-related proteins P62 and mitochondrial outer membrane transposase 20(TOMM20)decreased in T2DM group.Compared with DCI group,the cognitive dysfunction of mice improved,and the expression level of LncRNA SNHG16 decreased in the DCI+Mdivi-1 group(P<0.05).The expressions of LncRNA SNHG16,LC3 Ⅱ/Ⅰ,PINK1 and Parkin were higher in HG group than in Con group(P<0.05),while the cell survival rate and TOMM20 protein expression were lower in HG group than in Con group(P<0.05).Silence of LncRNA SNHG16 can restore the activity of HT22 cells and mitochondrial membrane potential,and reduce the level of mitochondrial autophagy under HG condition.Conclusions The expression level of LncRNA SNHG16 was up-regulated in the hippocampus brain region of mice with diabetic cognitive dysfunction,and mitophagy was overactivated.Silencing of LncRNA SNHG16 inhibits mitophagy in hippocampal neurons and alleviates HG induced hippocampal neuronal damage.

Objective To investigate the impact of mitophagy,mediated by the long non-coding RNA SNHG16(LncRNA SNHG16)on diabetes-associated cognitive impairment(DCI).Methods 29 male C57BL/J mice were randomly divided into normal control(NC)group,DCI group and DCI+mitochondrial autophagy inhibitor(DCI+Mdivi-1)group.Morris water maze and new object recognition test were used to detect the cognitive function of mice,qRT-CPR was used to detect the expression of LncRNA SNHG16 and mitochondrial autophagy marker mRNA.Western blot were used to detect the expression of related protein.The mouse hippocampal neurons HT22 were divided into control(Con)group,high glucose(HG)group,HG+SNHG16 silencing(HG+sh-SNHG16)group and HG+no-load control(HG+sh-NC)group.CCK8 method and lactate dehydrogenase(LDH)method were used to detect neuronal damage.JC-1 method was used to detect mitochondrial membrane potential.Results Compared with NC group,the expression of LncRNA SNHG16 and the expression of autophagy-related gene 5,PTEN-induced putative kinase 1(PINK1),Parkin and microtubule associated protein light chain 3(LC3)Ⅱ/Ⅰ increased(P<0.05),while the expression of mitochondrial autophagy-related proteins P62 and mitochondrial outer membrane transposase 20(TOMM20)decreased in T2DM group.Compared with DCI group,the cognitive dysfunction of mice improved,and the expression level of LncRNA SNHG16 decreased in the DCI+Mdivi-1 group(P<0.05).The expressions of LncRNA SNHG16,LC3 Ⅱ/Ⅰ,PINK1 and Parkin were higher in HG group than in Con group(P<0.05),while the cell survival rate and TOMM20 protein expression were lower in HG group than in Con group(P<0.05).Silence of LncRNA SNHG16 can restore the activity of HT22 cells and mitochondrial membrane potential,and reduce the level of mitochondrial autophagy under HG condition.Conclusions The expression level of LncRNA SNHG16 was up-regulated in the hippocampus brain region of mice with diabetic cognitive dysfunction,and mitophagy was overactivated.Silencing of LncRNA SNHG16 inhibits mitophagy in hippocampal neurons and alleviates HG induced hippocampal neuronal damage.

Objective:To establish and optimize an autoverification system for thyroid function test reports of 5 items using historical test data.Methods:Based on the docoment' Autoverification of Clinical Laboratory Quantitative Test Results′, CLSI AUTO 10-A and AUTO 15 guidelines, an autoverification system for thyroid function test reports of 5 items was established combining with manual verification experience. A total of 193 860 thyroid function test reports of 5 items in 2021 were collected for the assessment of the original system. Totally 210 097 thyroid function test reports of 5 items in 2022 and 299 198 reports in 2023 were collected for the optimization of the autoverification system. There were 160 666 thyroid function test reports of 5 items from the first half of 2024 for the manual and autoverification comparison after optimization.Results:The pass rate of the autoverification system based on original thyroid function report in 2021 was 69.56%(134 849/193 860). The optimized system utilizing historical data from 2022 and 2023 covered 21 pattern rules and established verification for different patterns including range rules, delta check rules, and review rules. Taking manual verification as the standard for the data from the first half of 2024, the sensitivity and specificity of the optimized system were 100% (499/499) and 81.57% (130 646/160 167), respectively, with a false-negative rate of 0. The concordance rate between autoverification and manual verification was 81.63% (131 145/160 666), and the pass rate was 81.32% (130 646/160 666).Conclusion:Establishing and optimizing the autoverification system for thyroid function tests of 5 items using historical test data, and formulating verification rules for different patterns can be applied to clinical practise, which not only ensures the accuracy of test reports but also improves work efficiency, allowing continuously optimized and perfected of the system.

Objective:To construct a biosafety evaluation index system for major emergency public health events in medical institutions.Methods:Based on previous laboratory biosafety evaluation work, relevant regulations and standards on biosafety in China were collected through literature research and expert consultations. Candidate indicators for constructing the biosafety evaluation system for major emergency public health events in medical institutions were selected, and a framework was established. Two rounds of expert questionnaires were conducted to determine the content of the index system based on experts′ evaluation, and each indicator′s relevance and importance were scored. Finally, two rounds of Delphi consultations were carried out, and the Analytic Hierarchy Process (AHP) was applied to calculate the weights of indicators.Results:The response rates for the total four rounds of questionnaire surveys were all 100%. The first two rounds focused on determining the framework, while the latter two focused on determining the weights for each indicator. The authority coefficients of the expert consultations for the two rounds of weights were 0.65 and 0.70, respectively, indicating the reliability of the research results. In the final round of survey, the Kendall′s coefficients of concordance at each level were all greater than 0.1. Through statistical testing, the P-values were all less than 0.05, indicating good coordination of expert opinions. Ultimately, we established an operational biosafety evaluation system for major emergency public health events in medical institutions, consisting of 4 primary indicators, 26 secondary indicators, and 119 tertiary indicators, with additional deduction items, bonus items, unacceptable items, and monitoring indicators.Conclusions:Based on scientific theory, a biosafety evaluation system for major emergency public health events in medical institutions was constructed, achieving the integration of routine and emergency measures. This system can be used for self-assessment of laboratory biosafety during emergency public health events, addressing the lack of unified standards in biosafety evaluation. Through regular self-assessment, it can enhance the level of biosafety management in medical institution laboratories, to realize the value of application and dissemination.

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 establish and optimize an autoverification system for thyroid function test reports of 5 items using historical test data.Methods:Based on the docoment' Autoverification of Clinical Laboratory Quantitative Test Results′, CLSI AUTO 10-A and AUTO 15 guidelines, an autoverification system for thyroid function test reports of 5 items was established combining with manual verification experience. A total of 193 860 thyroid function test reports of 5 items in 2021 were collected for the assessment of the original system. Totally 210 097 thyroid function test reports of 5 items in 2022 and 299 198 reports in 2023 were collected for the optimization of the autoverification system. There were 160 666 thyroid function test reports of 5 items from the first half of 2024 for the manual and autoverification comparison after optimization.Results:The pass rate of the autoverification system based on original thyroid function report in 2021 was 69.56%(134 849/193 860). The optimized system utilizing historical data from 2022 and 2023 covered 21 pattern rules and established verification for different patterns including range rules, delta check rules, and review rules. Taking manual verification as the standard for the data from the first half of 2024, the sensitivity and specificity of the optimized system were 100% (499/499) and 81.57% (130 646/160 167), respectively, with a false-negative rate of 0. The concordance rate between autoverification and manual verification was 81.63% (131 145/160 666), and the pass rate was 81.32% (130 646/160 666).Conclusion:Establishing and optimizing the autoverification system for thyroid function tests of 5 items using historical test data, and formulating verification rules for different patterns can be applied to clinical practise, which not only ensures the accuracy of test reports but also improves work efficiency, allowing continuously optimized and perfected of the system.

Objective:To construct a biosafety evaluation index system for major emergency public health events in medical institutions.Methods:Based on previous laboratory biosafety evaluation work, relevant regulations and standards on biosafety in China were collected through literature research and expert consultations. Candidate indicators for constructing the biosafety evaluation system for major emergency public health events in medical institutions were selected, and a framework was established. Two rounds of expert questionnaires were conducted to determine the content of the index system based on experts′ evaluation, and each indicator′s relevance and importance were scored. Finally, two rounds of Delphi consultations were carried out, and the Analytic Hierarchy Process (AHP) was applied to calculate the weights of indicators.Results:The response rates for the total four rounds of questionnaire surveys were all 100%. The first two rounds focused on determining the framework, while the latter two focused on determining the weights for each indicator. The authority coefficients of the expert consultations for the two rounds of weights were 0.65 and 0.70, respectively, indicating the reliability of the research results. In the final round of survey, the Kendall′s coefficients of concordance at each level were all greater than 0.1. Through statistical testing, the P-values were all less than 0.05, indicating good coordination of expert opinions. Ultimately, we established an operational biosafety evaluation system for major emergency public health events in medical institutions, consisting of 4 primary indicators, 26 secondary indicators, and 119 tertiary indicators, with additional deduction items, bonus items, unacceptable items, and monitoring indicators.Conclusions:Based on scientific theory, a biosafety evaluation system for major emergency public health events in medical institutions was constructed, achieving the integration of routine and emergency measures. This system can be used for self-assessment of laboratory biosafety during emergency public health events, addressing the lack of unified standards in biosafety evaluation. Through regular self-assessment, it can enhance the level of biosafety management in medical institution laboratories, to realize the value of application and dissemination.

Objective:To evaluate the viability and clinical effect of polyfoliate anterolateral thigh perforator flap (ALTPF) in reconstruction of large soft tissue defect around ankle.Methods:From June 2019 to October 2022, large soft tissue defects around ankle of 11 patients were reconstructed with ALTPF in the Department of Orthopaedics of the First Affiliated Hospital of Nanchang University. The causes of injury were traffic accident in 8 patients and heavy objects in 3 patients. All wounds were large defects (15.0 cm×14.0 cm-30.0 cm×20.0 cm) and combined with various degrees of infection. Intraoperatively, polyfoliate ALTPFs sized 16.0 cm×14.5 cm-23.0 cm×18.5 cm were used in reconstruction of the defects. Deep dead spaces were filled with antibiotic bone cement, and direct suture was performed to close the donor sites or by skin grafting repair. Postoperative follow-ups were scheduled at 1, 3 and 6 months, and 6 monthly thereafter at outpatient clinics and via telephone interviews. The appearance and colour of the flaps and healing of donor sites were recorded together with evaluation of the recovery of ankle motor function according to the ankle-hindfoot rating scale of American Orthopaedic Foot and Ankle Society (AOFAS).Results:All flaps survived. No haematoma or secondary infection occurred at the recipient site after surgery. All donor sites healed primarily. One patient had venous occlusion at the distal end of the polyfoliate ALTPF. The flap survived completely at 1 week after distal venous bloodletting. Postoperative follow-ups lasted 6-24 (15.27±5.21) months. All flaps had good blood supply with satisfactory appearance, similar colour and texture to the recipient sites, and without obvious bloat nor ulceration. Only a linear scar or few skin graft scar was left at the flap donor sites in concealed locations. The mean AOFAS ankle-hindfoot score was (88.36±10.21) point. There were 6 cases of excellent, 4 cases of good, and 1 case of fair.Conclusion:A polyfoliate ALTPF is an ideal flap for reconstruction of soft tissue defects around ankle by converting the length of a flap to the width.

Objective To compare and analyze the efficacy and safety of 5-needle injection and standard 20-needle injection in the treatment of overactive bladder with botulinum toxin type A.Methods A retrospective analysis was performed on 48 patients with overactive bladder who received intravesical injection of botulinum toxin type A in the Department of Urology,Hangzhou Third People's Hospital from January 2015 to December 2022,and they were divided into two groups according to the number of injections,with 24 patients in each group.The observation group received 5-needle injection,and the control group received standard 20-needle injection.Average daily frequency of urination,international consultation on incontinence questionnaire-overactive bladder,international consultation on incontinence questionnaire-overactive bladder(ICIQ-OAB)score,overactive bladder(OAB-Q)score,visual analogue scale(VAS),patient generated index(PGI-I)score,complication rate and willingness to repeat injection were recorded before and after treatment in two groups,respectively.Results There were no significant differences in age,gender,course of disease,average daily frequency of urination before treatment and baseline data of each score between the two groups,which were comparable.All patients completed treatment,and compared with before treatment,the mean daily frequency of urination,ICIQ-OAB and OAB-Q were improved after treatment(P<0.05),there was no significant difference between the two groups(P>0.05).There were no significant differences in scores and incidence of complications between the two groups after treatment(P>0.05).However,patients in the observation group were more willing to receive another injection(P<0.05).Conclusion The efficacy and safety of 5-needle vesical injection of botulinum toxin type A in the treatment of overactive bladder is similar to that of standard 20-needle injection,which is more easily accepted by patients,and is a safe and effective alternative to standard technique.