1.Mechanism of Kidney-tonifying and Liver-regulating Cyclical Therapy and Formula in Improving Endometrial Receptivity during "Implantation Window" in Rats with Polycystic Ovary Syndrome via miR-140-5p/VEGF Pathway
Quan LIU ; Yiqing HE ; Ying ZHANG ; Xi WANG ; Shuo YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):100-109
		                        		
		                        			
		                        			ObjectiveTo investigate the mechanism of kidney-tonifying and liver-regulating cyclical therapy and its formula in regulating endometrial receptivity during the "implantation window" in rats with polycystic ovary syndrome (PCOS). MethodsSix rats were randomly selected from 36 SPF SD female rats as the normal group, and the remaining rats were administered letrozole to induce a PCOS model. By using a random number method, the rats were divided into the following groups: normal group, model group, Xiaoyaosan group (11.97 g·kg-1), Sanzi Yangmo decoction group (28.35 g·kg-1), cyclical therapy group (11.97/28.35 g·kg-1), and aspirin group (8 × 10-3 mg·kg-1). After 12 days of continuous administration by gavage (equivalent to three estrous cycles), female and male rats were co-housed. On the fifth day of pregnancy, the number of blastocyst implantation in each group was counted. Hematoxylin-eosin (HE) staining was used to observe the pathological morphology of rat endometrial tissue. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of estradiol (E2) and progesterone (P) in rat serum. Western blot was used to detect the protein expression of vascular endothelial growth factor (VEGF), progesterone receptor (PR), estrogen receptor (ER), androgen receptor (AR), and integrin(ITG) αvβ3 in rat endometrial blood vessels. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of miR-140-5 P, VEGF, vascular endothelial growth factor receptor 2 (VEGFR2), PR, ER, AR, and ITGαvβ3 in rat endometrium. ResultsCompared with normal group, the estrous cycle of the rats in model group continued to be in the estrus interval and the estrous cycle lost regular changes. The endometrium was significantly thinner, the number of uterine glands and blood vessels were significantly reduced (P<0.01), and the pregnancy rate was significantly reduced. Compared with the model group, each drug group restored the regular estrous cycle to varying degrees, and the endometrial thickness and the number of blood vessels were significantly improved (P<0.01). The pregnancy rate of each drug group increased, and the effect of the cycle therapy group could reach the normal level. The results of molecular biology experiments showed that compared with the normal group, the levels of serum E2 and P in the model group were significantly decreased (P<0.01), the expression of VEGF, ER, PR and ITGαvβ3 protein was significantly decreased (P<0.05,P<0.01), the expression of AR protein was significantly increased (P<0.01), the expression of miR-140-5P and AR mRNA was significantly increased (P<0.01), and the expression of VEGF, VEGFR2, ER, PR and ITGαvβ3 mRNA was significantly decreased (P<0.01). Compared with model group, the serum E2 level in the Xiaoyaosan group was significantly increased (P<0.01).The levels of E2 and P in serum of rats in Sanzi Yangmo decoction group, cycle therapy group and aspirin group were significantly increased (P<0.01). The expression of AR protein in each drug group was significantly decreased (P<0.01). The expression of VEGF and ITGαvβ3 protein in Xiaoyaosan group was significantly increased (P<0.01). The expression of VEGF, ER and PR protein in Sanzi Yangmo decoction group was increased to varying degrees (P<0.05,P<0.01). The expression of VEGF, PR, ER and ITGαvβ3 protein in the cycle therapy group and the aspirin group increased to varying degrees (P<0.05,P<0.01). The expression of miR-140-5P and AR mRNA in each drug group was significantly decreased (P<0.01). The expression of VEGF, VEGFR2, ER, PR and ITGαvβ3 mRNA in each drug group increased to varying degrees (P<0.05,P<0.01). Compared with Xiaoyaosan group and Sanzi Yangmo decoction group, the expression of miR-140-5P, VEGFR2, ER, PR, AR and ITGαvβ3 mRNA in the cycle therapy group were significantly different (P<0.05,P<0.01). ConclusionThe kidney-tonifying and liver-regulating cyclical therapy may reduce the activity of miR-140-5P, target the upregulation of VEGF expression, mediate angiogenesis, and promote endometrial angiogenesis, thereby playing a synergistic role in improving endometrial receptivity in PCOS-induced infertility. Its efficacy in increasing pregnancy rates surpasses that of Xiaoyaosan or Sanzi Yangmo decoction used alone. 
		                        		
		                        		
		                        		
		                        	
2.Mechanism of Kidney-tonifying and Liver-regulating Cyclical Therapy and Formula in Improving Endometrial Receptivity during "Implantation Window" in Rats with Polycystic Ovary Syndrome via miR-140-5p/VEGF Pathway
Quan LIU ; Yiqing HE ; Ying ZHANG ; Xi WANG ; Shuo YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):100-109
		                        		
		                        			
		                        			ObjectiveTo investigate the mechanism of kidney-tonifying and liver-regulating cyclical therapy and its formula in regulating endometrial receptivity during the "implantation window" in rats with polycystic ovary syndrome (PCOS). MethodsSix rats were randomly selected from 36 SPF SD female rats as the normal group, and the remaining rats were administered letrozole to induce a PCOS model. By using a random number method, the rats were divided into the following groups: normal group, model group, Xiaoyaosan group (11.97 g·kg-1), Sanzi Yangmo decoction group (28.35 g·kg-1), cyclical therapy group (11.97/28.35 g·kg-1), and aspirin group (8 × 10-3 mg·kg-1). After 12 days of continuous administration by gavage (equivalent to three estrous cycles), female and male rats were co-housed. On the fifth day of pregnancy, the number of blastocyst implantation in each group was counted. Hematoxylin-eosin (HE) staining was used to observe the pathological morphology of rat endometrial tissue. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of estradiol (E2) and progesterone (P) in rat serum. Western blot was used to detect the protein expression of vascular endothelial growth factor (VEGF), progesterone receptor (PR), estrogen receptor (ER), androgen receptor (AR), and integrin(ITG) αvβ3 in rat endometrial blood vessels. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of miR-140-5 P, VEGF, vascular endothelial growth factor receptor 2 (VEGFR2), PR, ER, AR, and ITGαvβ3 in rat endometrium. ResultsCompared with normal group, the estrous cycle of the rats in model group continued to be in the estrus interval and the estrous cycle lost regular changes. The endometrium was significantly thinner, the number of uterine glands and blood vessels were significantly reduced (P<0.01), and the pregnancy rate was significantly reduced. Compared with the model group, each drug group restored the regular estrous cycle to varying degrees, and the endometrial thickness and the number of blood vessels were significantly improved (P<0.01). The pregnancy rate of each drug group increased, and the effect of the cycle therapy group could reach the normal level. The results of molecular biology experiments showed that compared with the normal group, the levels of serum E2 and P in the model group were significantly decreased (P<0.01), the expression of VEGF, ER, PR and ITGαvβ3 protein was significantly decreased (P<0.05,P<0.01), the expression of AR protein was significantly increased (P<0.01), the expression of miR-140-5P and AR mRNA was significantly increased (P<0.01), and the expression of VEGF, VEGFR2, ER, PR and ITGαvβ3 mRNA was significantly decreased (P<0.01). Compared with model group, the serum E2 level in the Xiaoyaosan group was significantly increased (P<0.01).The levels of E2 and P in serum of rats in Sanzi Yangmo decoction group, cycle therapy group and aspirin group were significantly increased (P<0.01). The expression of AR protein in each drug group was significantly decreased (P<0.01). The expression of VEGF and ITGαvβ3 protein in Xiaoyaosan group was significantly increased (P<0.01). The expression of VEGF, ER and PR protein in Sanzi Yangmo decoction group was increased to varying degrees (P<0.05,P<0.01). The expression of VEGF, PR, ER and ITGαvβ3 protein in the cycle therapy group and the aspirin group increased to varying degrees (P<0.05,P<0.01). The expression of miR-140-5P and AR mRNA in each drug group was significantly decreased (P<0.01). The expression of VEGF, VEGFR2, ER, PR and ITGαvβ3 mRNA in each drug group increased to varying degrees (P<0.05,P<0.01). Compared with Xiaoyaosan group and Sanzi Yangmo decoction group, the expression of miR-140-5P, VEGFR2, ER, PR, AR and ITGαvβ3 mRNA in the cycle therapy group were significantly different (P<0.05,P<0.01). ConclusionThe kidney-tonifying and liver-regulating cyclical therapy may reduce the activity of miR-140-5P, target the upregulation of VEGF expression, mediate angiogenesis, and promote endometrial angiogenesis, thereby playing a synergistic role in improving endometrial receptivity in PCOS-induced infertility. Its efficacy in increasing pregnancy rates surpasses that of Xiaoyaosan or Sanzi Yangmo decoction used alone. 
		                        		
		                        		
		                        		
		                        	
3.Methodological Evaluation of Advantages of Traditional Chinese Medicine Treatment of Sjögren's Syndrome
Wenjing LIU ; Shiya WU ; Ruihua LIU ; Xinyao ZHOU ; Juan JIAO ; Ying LIU ; Zeguang LI ; Zhenbin LI ; Huadong ZHANG ; Xiaopo TANG ; Quan JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(1):192-197
		                        		
		                        			
		                        			Screening and evaluating the diseases responding specifically to traditional Chinese medicine (TCM) will help to highlight the advantages of TCM treatment, and the evaluation method should be standardized with consideration to the unique characteristics of the diseases. The incidence of Sjögren's Syndrome (SS) is increasing year by year, while the pathogenesis of this disease remains unclear. Modern therapies for this disease include biological agents and immunosuppressants, which generally have unsatisfactory efficacy. The TCM treatment of SS focuses on the harmony of the physical and mental health. The Rheumatology Branch of the China Association of Chinese Medicine organizes experts in TCM, Western medicine, and evidence-based medicine to form working groups. Delphi method and bibliometric method were used for analysis, and SS was selected as a disease responding specifically to TCM. Furthermore, the evaluation system was established for this disease, and the consensus regarding this disease was reached after seminar discussion. This paper summarized the whole process of the evaluation of the advantages of TCM treatment of SS. First, because TCM atomization is widely used in clinical practice and enriches TCM administration methods, this therapy is included after other non-drug therapies were taken as characteristic therapies. Second, the evaluation indicators of therapeutic effect should be determined with consideration to international acceptance and the current research status. Third, the expression method should be accurate, standardized, and objective, highlight the natural advantages of TCM, and avoid arbitrary extension. This paper provides a reference for clinicians to explore other diseases responding specifically to TCM. 
		                        		
		                        		
		                        		
		                        	
4.National bloodstream infection bacterial resistance surveillance report (2022) : Gram-negative bacteria
Zhiying LIU ; Yunbo CHEN ; Jinru JI ; Chaoqun YING ; Qing YANG ; Haishen KONG ; Haifeng MAO ; Hui DING ; Pengpeng TIAN ; Jiangqin SONG ; Yongyun LIU ; Jiliang WANG ; Yan JIN ; Yuanyuan DAI ; Yizheng ZHOU ; Yan GENG ; Fenghong CHEN ; Lu WANG ; Yanyan LI ; Dan LIU ; Peng ZHANG ; Junmin CAO ; Xiaoyan LI ; Dijing SONG ; Xinhua QIANG ; Yanhong LI ; Qiuying ZHANG ; Guolin LIAO ; Ying HUANG ; Baohua ZHANG ; Liang GUO ; Aiyun LI ; Haiquan KANG ; Donghong HUANG ; Sijin MAN ; Zhuo LI ; Youdong YIN ; Kunpeng LIANG ; Haixin DONG ; Donghua LIU ; Hongyun XU ; Yinqiao DONG ; Rong XU ; Lin ZHENG ; Shuyan HU ; Jian LI ; Qiang LIU ; Liang LUAN ; Jilu SHEN ; Lixia ZHANG ; Bo QUAN ; Xiaoping YAN ; Xiaoyan QI ; Dengyan QIAO ; Weiping LIU ; Xiusan XIA ; Ling MENG ; Jinhua LIANG ; Ping SHEN ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2024;17(1):42-57
		                        		
		                        			
		                        			Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of national bloodstream infection Bacterial Resistant Investigation Collaborative System(BRICS)were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute(CLSI). WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:During the study period,9 035 strains of Gram-negative bacteria were collected from 51 hospitals,of which 7 895(87.4%)were Enterobacteriaceae and 1 140(12.6%)were non-fermenting bacteria. The top 5 bacterial species were Escherichia coli( n=4 510,49.9%), Klebsiella pneumoniae( n=2 340,25.9%), Pseudomonas aeruginosa( n=534,5.9%), Acinetobacter baumannii complex( n=405,4.5%)and Enterobacter cloacae( n=327,3.6%). The ESBLs-producing rates in Escherichia coli, Klebsiella pneumoniae and Proteus spp. were 47.1%(2 095/4 452),21.0%(427/2 033)and 41.1%(58/141),respectively. The prevalence of carbapenem-resistant Escherichia coli(CREC)and carbapenem-resistant Klebsiella pneumoniae(CRKP)were 1.3%(58/4 510)and 13.1%(307/2 340);62.1%(36/58)and 9.8%(30/307)of CREC and CRKP were resistant to ceftazidime/avibactam combination,respectively. The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)complex was 59.5%(241/405),while less than 5% of Acinetobacter baumannii complex was resistant to tigecycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa(CRPA)was 18.4%(98/534). There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of main Gram-negative bacteria resistance among different regions,with statistically significant differences in the prevalence of CRKP and CRPA( χ2=20.489 and 20.252, P<0.001). The prevalence of CREC,CRKP,CRPA,CRAB,ESBLs-producing Escherichia coli and Klebsiella pneumoniae were higher in provinicial hospitals than those in municipal hospitals( χ2=11.953,81.183,10.404,5.915,12.415 and 6.459, P<0.01 or <0.05),while the prevalence of CRPA was higher in economically developed regions(per capita GDP ≥ 92 059 Yuan)than that in economically less-developed regions(per capita GDP <92 059 Yuan)( χ2=6.240, P=0.012). Conclusions:The proportion of Gram-negative bacteria in bloodstream infections shows an increasing trend,and Escherichia coli is ranked in the top,while the trend of CRKP decreases continuously with time. Decreasing trends are noted in ESBLs-producing Escherichia coli and Klebsiella pneumoniae. Low prevalence of carbapenem resistance in Escherichia coli and high prevalence in CRAB complex have been observed. The composition ratio and antibacterial spectrum of bloodstream infections in different regions of China are slightly different,and the proportion of main drug resistant bacteria in provincial hospitals is higher than those in municipal hospitals.
		                        		
		                        		
		                        		
		                        	
5.Liuwei Dihuang Pills-elicited inhibition of MMP-2/MMP-9 via RAGE on tight junction protein of Aβ1-40-injured bEnd.3 cells
Rui DING ; Yong YUAN ; Ya-Quan JIA ; Ai-She GAO ; Zhen-Qiang ZHANG ; Jun-Ying SONG
Chinese Traditional Patent Medicine 2024;46(2):424-430
		                        		
		                        			
		                        			AIM To investigate the protective effects and the mechanism of the Liuwei Dihuang Pills on mouse brain microvascular endothelial(bEnd.3)cells damaged by β-Amyloid protein1-40(Aβ1-40).METHODS CCK8 method was used to detect the effects of Aβ1-40 and medicated serum of Liuwei Dihuang Pills(MSLDP)on cell activity,and to screen the appropriate concentration.bEnd.3 cells of the control group,the Aβ1-40 group,the MSLDP+Aβ1-40 group and the MSLDP group had their low density lipoprotein-associated protein 1(LRP1),receptor for advanced glycation end products(RAGE),matrix metalloproteinase-2(MMP-2),MMP-9,scaffold protein zonule protein-1(ZO-1)detected by Western blot.bEnd.3 cells assigned into the control group,the Aβ1-40 group,the FPS-ZM1(RAGE inhibitor)+Aβ1-40 group and the FPS-ZM1+Aβ1-40+MSLDP group had their expressions of RAGE,MMP-9,MMP-2 and ZO-1 detected by Western blot as well.RESULTS The cell activity of bEnd.3,was dose-dependently decreased by Aβ1-40(P<0.01),but was protected by MSLDP(P<0.05,P<0.01).And 10 μmol/L Aβ1-40 and 10%MSLDP were selected for subsequent experiments.Compared with the control group,the Aβ1-40 group displayed increased protein expressions of RAGE,MMP-2 and MMP-9(P<0.01),decreased protein expressions of LRP1,ZO-1 and BDNF(P<0.05,P<0.01),and decreased fluorescence intensities of LRP1 and ZO-1(P<0.01).Compared with the Aβ1-40 group,the MSLDP group shared decreased expressions of RAGE,MMP-2,MMP-9 proteins and RAGE fluorescence intensity(P<0.05,P<0.01),and increased expressions of LRP1,ZO-1 and BDNF proteins,and the fluorescence intensity of LRP1 and ZO-1(P<0.05,P<0.01);the Aβ1-40+FPS-ZM1 group displayed decreased protein expressions of MMP-2,MMP9 and RAGE(P<0.05,P<0.01),and increased ZO-1 protein expression(P<0.05);and the Aβ1-40+FPS-ZM1+ MSLDP group displayed an even more decreased protein expressions of MMP-2,MMP9 and RAGE(P<0.01),increased ZO-1 protein expression(P<0.01)due to the the combination use of FPS-ZM1 and MSLDP.CONCLUSION Liuwei Dihuang Pills can protect the tight junction of bEnd.3 injured by Aβ1-40 and neurovascular units from Alzheimer's disease by alleviating the dysfunction of the blood-brain barrier via RAGE-mediated MMP-2/MMP-9 pathway inhibition.
		                        		
		                        		
		                        		
		                        	
6.Preparation and Recognition Features of Molecularly Imprinted Polymer Membrane for Lamotrigine in Plasma
Dong-Yu LU ; Yu-Xin YOU ; Yan-Lin ZHAO ; Han JIANG ; Ying ZHANG ; Yan DU ; Dao-Quan TANG
Chinese Journal of Analytical Chemistry 2024;52(1):80-92
		                        		
		                        			
		                        			The molecularly imprinted polymers membranes(MIPMs)were prepared for selective adsorption of lamotrigine(LTG)in plasma by surface molecular imprinting technology with polyvinylidenefluoride(PVDF)membranes as supporter,lamotrigine as template molecule,methyl methacrylate as functional monomer,ethylene glycol dimethacrylate as cross-linking agent,azodiisobutyronitrile as initiator and acetonitrile-dimethylformamide(1∶1.5,V/V)as pore-forming agent.The prepared MIPMs were characterized by scanning electron microscope,Fourier transform infrared spectroscopy,Brunaner-emmet-teller measurements,X-ray photoelectron spectroscopy,and thermogravimetric analysis.The adsorption properties of the materials were investigated by kinetic adsorption,isothermal adsorption,selective adsorption,adsorption-desorption and reusability experiments.The results showed that the imprinted layer of LTG was successfully coated on the surface of PVDF,and the materials had uniform particle size.The adsorption capacity and imprinting factor of the MIPMs towards LTG were 3.77 mg/g and 8.97,respectively.The nanomaterials showed fast mass transfer rate(30 min)and good reusability(the adsorption efficiency was 86.66%after 6 cycles),and could be used for the adsorption of LTG in plasma with low matrix interference,recoveries of 86.54%-90.48%and RSD of 1.51%-3.15%(n=5).The proposed LTG MIPMs were demonstrated to be simple and environment friendly,and had high selectivity in rapid separation and extraction of LTG in plasma.
		                        		
		                        		
		                        		
		                        	
7.Treatment of Neer Ⅱ b distal clavicular fracture with locking plate under arthroscope
Shao CHEN ; Cai-Hua ZHANG ; Quan-Ming YANG ; Xin-Chun LI ; Ying-Chun ZHU
China Journal of Orthopaedics and Traumatology 2024;37(10):1010-1014
		                        		
		                        			
		                        			Objective To investigate the feasibility and clinical effect of the treatment of Neer Ⅱ b distal clavicular fracture with locking plate under arthroscopy.Methods Between June 2018 and September 2022,the medical records of 15 patients with Neer Ⅱ b distal clavicular fracture treated with locking plate under arthroscope were retrospectively analyzed,including 9 males and 6 females,aged from 25 to 57 years old with an average of(42.50±7.75)years old,5 left shoulder,10 right shoul-der,and duration of the disease ranged from 4 to 7 days with an average of(5.16±0.98)d.Visual analog score(VAS),Con-stant-Murley shoulder joint score scale and Neer standard score were used before operation,1 month after operation and the last follow-up.Results All patients successfully completed the treatment of Neer Ⅱ b distal clavicular fracture with locking plate under arthroscopy,with the operation time ranging from 0.3 to 1.0 h with an average of(0.55±0.24)h.All patients were healed at stage Ⅰ.The follow-up time ranged from 6 to 15 months with an average of(9.2±2.8)months.The preoperative VAS score was 7.23±0.67,the Constant-Murley shoulder joint score was(19.57±0.91),and the Neer standard score was(11.27±1.12);The VAS score was(1.56±0.81),the Constant-Murley shoulder joint score was(52.62±1.54),and the Neer standard score was(61.98±2.99)in the first month after operation;At the last follow-up,the VAS was(0.42±0.54),the Constant-Murley shoulder joint score was(91.24±1.97),and the Neer standard score was(93.24±3.38).The difference between VAS,Con-stant-Murley shoulder joint score and Neer standard score was statistically significant at one month postoperative and preoper-ative(P<0.05),and the difference between the last follow-up visit and one month postoperative was statistically significant(P<0.05).Conclusion The treatment of Neer Ⅱ b distal clavicular fracture with locking plate under arthroscope can restore the function of shoulder joint,with definite curative effect and feasibility.
		                        		
		                        		
		                        		
		                        	
8.Evaluation of the Effect for National Centralized Drug Procurement Policy of PPIs in 33 Hospitals in Wuhan
Zhijuan LIN ; Li LIU ; Wenjuan HE ; Zhiliang ZHANG ; Zhaohui GUO ; Ping LIU ; Quan LEI ; Ying XU
Chinese Journal of Modern Applied Pharmacy 2024;41(12):1723-1728
		                        		
		                        			OBJECTIVE 
		                        			To analyze the use status and development trend of proton pump inhibitors(PPIs) in 33 hospitals in Wuhan, Hubei Province after the implementation of the national centralized drug procurement(NCDP) policy, and to provide reference for promoting the subsequent rational use of NCDP drugs and improving related policies.
METHODS 
To make statistics and analysis of purchasing amount of PPIs, defined daily dose system(DDDs), defined daily dose consumption(DDDc) and utilization rate of 33 hospitals in Wuhan in 2019 and 2022.
RESULTS 
After the implementation of the NCDP policy, the total purchasing amount of PPIs decreased by 53.6%, DDDs decreased by 15.4%, DDDc decreased by 45.2%, and the utilization rate of PPIs injectable dosage forms decreased by 12.6%. After NCDP, the highest growth rate of oral dosage forms was omeprazole(5.7%), followed by rabeprazole(5.0%), while injectable dosage forms showed a significant difference in utilization rate, with a significant decline in NCDP varieties and a significant increase in non-NCDP varieties. The overall NCDP utilization rate of PPIs in Wuhan was 64.9%, with little difference among hospitals of different grades.
CONCLUSION 
The NCDP policy achieves the purpose of reducing the drug cost of patients and improving the accessibility of drugs, and is more optimized in the selection of dosage forms, which is in line with the policy expectations overall; but the quantity and price of PPIs in Wuhan decreased after NCDP, and highlighted a certain tendency in the selection of varieties. In the future, we still need to optimize measures to guide clinical priority in the selection of NCDP drugs, to ensure and improve the implementation of NCDP policy.
		                        		
		                        		
		                        		
		                        	
9.Disease characteristics and costs of pediatric Mycoplasma Pneumoniae pneumonia hospitalization:a retrospective study at municipal hospitals from 2019 to 2023 in Shanghai
Ying-Wen WANG ; Feng WANG ; Li-Bo WANG ; Ai-Zhen LU ; Yi WANG ; Yong-Hao GUI ; Quan LU ; Yong YIN ; Jian-Hua ZHANG ; Ying-Zi YE ; Hong XU ; Bing SHEN ; Dan-Ping GU ; Xiao-Yan DONG ; Jia-Yu WANG ; Wen HE ; Xiao-Bo ZHANG
Fudan University Journal of Medical Sciences 2024;51(4):515-521
		                        		
		                        			
		                        			Objective To investigate disease characteristics and hospitalization costs of children with Mycoplasma Pneumoniae pneumonia(MPP)admitted to Shanghai municipal medical hospitals from 2019 to 2023.Methods Depending on the Shanghai Municipal Hospital Pediatric Alliance,we retrospectively investigated community acquired MPP pediatric patients hospitalized in 22 municipal hospitals with pediatric qualifications(including 4 children's hospitals)in Shanghai from Jan 2019 to Dec 2023.We collected the patients'diagnosis codes,gender,age,length of hospital stay,hospitalization costs,and whether they progressed to severe Mycoplasma pneumoniae pneumonia(SMPP).Results From 2019 to 2023,a total of 29 045 hospitalized children with MPP were treated,with 6 035 cases(20.8%)identified as SMPP in the 22 hospitals.Trend analysis revealed a rising trend with years in the proportion of SMPP patients(χ2trend=365.498,P<0.001).Among the 4 children's hospitals,there were 18 710 cases with MPP,including 4 078 cases(21.8%)of SMPP.The proportion of SMPP patients also showed an increasing trend with years(χ2trend=14.548,P<0.001),and the proportion in 2023(23.0%)was higher than that in previous years with statistical significance.There were statistical differences in the seasonal distribution of MPP cases between different years,with higher proportions in summer and autumn overall.The age distribution of hospitalized MPP children varied among different years,with school-age children accounting for the majority(56.8%)in 2023.There was no difference in the distribution of severe cases between different genders,but there were differences in the proportion of severe cases among different age groups in different years,with a gradual increase in severe cases among children aged 1 to 3 years(χ2trend=191.567,P<0.001).The average length of hospital stay for MPP during the epidemic was higher than that during non-epidemic periods,and there were statistically significant differences in the average length of hospital stay between different years(P<0.001).The individual hospitalization costs during the epidemic were higher than in other years,and there were statistically significant differences in individual hospitalization costs between different years(P<0.001).The total hospitalization costs were still higher in 2019 and 2023.The individual hospitalization costs for SMPP were higher than for non-SMPP cases.Conclusion MPP outbreaks occurred in Shanghai in 2019 and 2023,with the higher proportions in summer and autumn overall.Compared to previous years,the number of hospitalized MPP children in Shanghai was higher in 2023,with a higher proportion of SMPP cases,especially among children under 3 years old.The individual per capita hospitalization expenses for SMPP cases were higher than for non-SMPP cases.
		                        		
		                        		
		                        		
		                        	
10.Imbalance of programmed cell death patterns mediated by dendritic cell subsets in systemic lupus erythematosus and lupus nephritis
Ruoyao XU ; Ying ZHANG ; Qingtai CAO ; Sheng LIAO ; Youzhou TANG ; Quan ZHUANG
Journal of Central South University(Medical Sciences) 2024;49(3):331-348
		                        		
		                        			
		                        			Objective:Abnormal programmed cell death in immune cells is associated with autoimmune diseases,but the patterns of programmed cell death in systemic lupus erythematosus(SLE)and especially lupus nephritis(LN)remain unclear.This study aims to explore the association between SLE,LN,and immune cell death patterns. Methods:Bulk RNA sequencing(bulk RNA-seq)and single-cell RNA sequencing(scRNA-seq)data were downloaded from the Gene Expression Omnibus(GEO)database.Bioinformatic analysis was conducted to explore the expression levels of genes related to 3 cell death patterns in peripheral blood mononuclear cells of SLE patients.Key cell subsets involved in the imbalance of cell death patterns were identified through scRNA-seq.Immunofluorescence was used to detect the expression levels of receptor interacting serine/threonine kinase 3(RIPK3),mixed-lineage kinase domain-like protein(MLKL),phosphorylated MLKL(pMLKL),caspase 1(CASP1),CD1c molecule(CD1C),C-type lectin domain containing 9A(CLEC9A),and X-C motif chemokine receptor 1(XCR1)in dendritic cells(DC).scRNA-seq was performed on kidney tissues collected from LN patients and healthy controls(HC)at the Third Xiangya Hospital of Central South University,followed by bioinformatic analysis to identify key cell subsets involved in the imbalance of cell death patterns.Pseudotime analysis and ligand-receptor analysis were used to explore the differentiation direction and cell communication of different DC subsets.Transient transfection was used to transfect RAW264.7 cells with empty plasmid,empty plasmid+dsDNA(HSV-DNA),empty plasmid+200 μmol/L tert-butyl hydroperoxide(TBHP),stimulator of interferon genes(STING)shRNA plasmid,STING shRNA plasmid+dsDNA(HSV-DNA),and STING shRNA plasmid+200 μmol/L TBHP.Annexin V-mCherry and SYTOX Green staining were used to detect cell death in each group.Western blotting was used to detect the activation of CASP1,gasdermin D(GSDMD),RIPK3,and MLKL in each group. Results:Bioinformatic analysis showed an imbalance in 3 cell death patterns in SLE and LN patients:Pro-inflammatory pyroptosis and necroptosis were activated,while anti-inflammatory apoptosis was inhibited.The key cell subsets involved were DC subsets,particularly focusing on CLEC9A+cDC1.Immunofluorescence results showed that the expression levels of RIPK3,MLKL,and CASP1 in DCs were higher in the SLE group compared to the HC group.pMLKL and CASP1 expression levels in renal cDC1 marked by CLEC9A and XCR1 were higher in the LN group than in the HC group.Pseudotime analysis and ligand-receptor analysis suggested that the CLEC9A+cDC1 subset in LN kidney tissues originated from peripheral circulation.Annexin V-mCherry and SYTOX Green staining results showed that the number of dead cells decreased in the STING shRNA transfection group compared to the empty plasmid group in RAW264.7 cells.Western blotting results showed that the activation of CASP1,GSDMD,RIPK3,and MLKL was decreased in the STING shRNA transfection group compared to the empty plasmid group. Conclusion:This study provides novel insights into the role of CLEC9A+cDC1 in the imbalance of cell death patterns in SLE and LN.
		                        		
		                        		
		                        		
		                        	
            

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