Objective To investigate the clinical distribution,antimicrobial resistance,carbapenemase resistance genes,virulence genes,capsular serotypes and ST subtypes of carbapenem-resistant Klebsiella pneumoniae(CRKP)strains in intensive care unit of a tertiary hospital in Hunan Province for better management of CRKP infections.Methods CRKP strains were isolated from 8 intensive care units of the First People's Hospital of Chenzhou City from January 2020 to December 2021.The isolates were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and tested by VITEK Compact 2 for antimicrobial susceptibility.Carbapenemase phenotype was detected by modified carbapenem inactivation method(mCIM).The capsular serotypes were determined by wzi sequencing.Carbapenem resistance genes and virulence genes were identified by PCR and Sanger sequencing.The strains were also analyzed by multilocus sequence typing(MLST)in terms of ST subtypes.Results The 75 CRKP strains were mainly isolated from geriatric ICU(28.0％)and neurosurgery ICU(20.0％).Overall,6.7％(5/75)and 16.0％(12/75)of the CRKP strains were resistant to tigecycline and ceftazidime-avibactam,respectively.The CRKP strains(＞96.0％resistant)were highly resistant to carbapenems,cephalosporins,β-lactam/β-lactamase inhibitor combinations,and levofloxacin.PCR and sequencing analysis found blaKPC-2 gene in 61 strains(81.3％),blaNDM-1 gene in 11 strains(14.7％),blaNDM-5 gene in 1 strain(1.3％),and blaOXA-48 gene in 2 strains(2.7％).MLST revealed that ST11(54.7％,41/75),ST1883(13.3％,10/75),and ST307(6.7％,5/75)were the top three ST subtypes.All ST11 and ST1883 CRKP strains harbored blaKPC-2.KL64(38.7％,29/75)and KL47(25.3％,19/75)were the most prevalent capsular serotypes among the 75 CRKP strains.The most common virulence genes among these CRKP strains were rmpA2(48.0％,36/75),iroN(38.7％,23/75)and iucA(37.3％,15/75).Conclusions The CRKP strains isolated from the intensive care units were mainly ST11-KL64 and ST11-KL47 types.Most of the strains harbor blaKPC-2 and virulence gene,and associated with high level antimicrobial resistance.It is urgent to strengthen the monitoring of molecular epidemiological characteristics of CRKP in order to inform individualized and precision treatment of CRKP infections.

Objective:To determine the expression of small nucleolar RNA host gene 16(SNHG16)in fibroblast-like synovio-cytes(FLS)of patients with rheumatoid arthritis(RA)and its role in the development of RA.Methods:RT-qPCR was performed to measure SNHG16,miR-182-5p and peroxisome proliferator-activated receptor gamma(PPARG)mRNA expression;dual-luciferase assay was performed to measure the interaction between SNHG16,miR-182-5p and PPARG mRNA;EdU and Transwell were per-formed to measure cell proliferation and invasion;Western blot was performed to measure PPARG protein level.Results:SNHG16 and PPARG mRNA expression were up-regulated and miR-182-5p expression was down-regulated in RA synovial tissue and human RA-FLS line(HFLS-RA).SNHG16 negatively targeted miR-182-5p expression and positively regulated PPARG(miR-182-5p target)expression.Silencing of SNHG16 inhibited the proliferation and invasion of HFLS-RA;down-regulation of miR-182-5p partially re-versed the inhibitory effect of SNHG16 silencing on cell proliferation and invasion;overexpression of PPARG partially reversed the in-hibitory effect of up-regulation of miR-182-5p on the proliferation and invasion of HFLS-RA.Conclusion:Silencing SNHG16 target-ing miR-182-5p/PPARG axis inhibits the proliferation and invasion of HFLS-RA.

OBJECTIVE To investigate the effects of ligustilide on chemotherapy resistance of cervical cancer cells based on Hippo-Yes-associated protein （YAP） signaling pathway. METHODS Human cervical cancer cisplatin-resistant cells HeLa/DDP were divided into control group， cisplatin group （10 μmol/L cisplatin）， cisplatin+ligustilide low-， medium- and high-concentration groups （10 μmol/L cisplatin+25， 50， 100 μmol/L ligustilide）. The proliferation， apoptosis， migration and invasion of HeLa/DDP cells were all detected in each group. The mRNA expressions of YAP and transcriptional coactivator with PDZ binding motif （TAZ） as well as the protein expressions of YAP， TAZ， matrix metalloproteinase 2 （MMP2）， Ki67， cleaved-caspase-3 and caspase-3 were determined in HeLa/DDP cells. RESULTS Compared with control group， the inhibitory rate， apoptotic rate and cleaved- caspase-3/caspase-3 of cisplatin group were increased significantly； scratch healing rate， the number of invasive cells， the mRNA expressions of YAP and TAZ， and the protein expressions of YAP， TAZ， MMP2 and Ki67 were decreased significantly in cisplatin group （P＜0.05）. Compared with cisplatin group， the inhibitory rate of cell proliferation， apoptotic rate and cleaved-caspase-3/ caspase-3 were further increased in cisplatin+ligustilide low-， medium- and high-concentration groups， while scratch healing rate， the number of invasive cells， the mRNA expressions of YAP and TAZ， and the protein expressions of YAP， TAZ， MMP2 and Ki67 were further decreased， in a dose-dependent manner （P＜0.05）. CONCLUSIONS Ligustilide can increase the sensitivity of drug-resistant cervical cancer cells to cisplatin by inhibiting Hippo-YAP signaling pathway.

Osteoporotic thoracolumbar fracture in the elderly will seriously reduce their quality of life and life expectancy. For osteoporotic thoracolumbar fracture in the elderly, spinal reconstruction is necessary, which should comprehensively consider factors such as the physical condition, fracture type, clinical characteristics and osteoporosis degree. While there lacks relevant clinical norms or guidelines on selection of spinal reconstruction strategies. In order to standardize the concept of spinal reconstruction for osteoporotic thoracolumbar fracture in the elderly, based on the principles of scientificity, practicality and progressiveness, the authors formulated the Clinical guideline for spinal reconstruction of osteoporotic thoracolumbar fracture in elderly patients ( version 2022), in which suggestions based on evidence of evidence-based medicine were put forward upon 10 important issues related to the fracture classification, non-operative treatment strategies and surgical treatment strategies in spinal reconstruction after osteoporosis thoracolumbar fracture in the elderly, hoping to provide a reference for clinical treatment.

Previous studies suggest that the reduction of SMAD3 (mothers against decapentaplegic homolog 3) has a great impact on tumor development, but its exact pathological function remains unclear. In this study, we found that the protein level of SMAD3 was greatly reduced in human-grade IV glioblastoma tissues, in which LAMP2A (lysosome-associated membrane protein type 2A) was significantly up-regulated. LAMP2A is a key rate-limiting protein of chaperone-mediated autophagy (CMA), a lysosome pathway of protein degradation that is activated in glioma. We carefully analyzed the amino-acid sequence of SMAD3 and found that it contained a pentapeptide motif biochemically related to KFERQ, which has been proposed to be a targeting sequence for CMA. In vitro, we confirmed that SMAD3 was degraded in either serum-free or KFERQ motif deleted condition, which was regulated by LAMP2A and interacted with HSC70 (heat shock cognate 71 kDa protein). Using isolated lysosomes, amino-acid residues 75 and 128 of SMAD3 were found to be of importance for this process, which affected the CMA pathway in which SMAD3 was involved. Similarly, down-regulating SMAD3 or up-regulating LAMP2A in cultured glioma cells enhanced their proliferation and invasion. Taken together, these results suggest that excessive activation of CMA regulates glioma cell growth by promoting the degradation of SMAD3. Therefore, targeting the SMAD3-LAMP2A-mediated CMA-lysosome pathway may be a promising approach in anti-cancer therapy.

Background::Parkinson’s disease (PD) is the second most common neurodegenerative disease after Alzheimer’s dementia. Mitochondrial dysfunction is involved in the pathology of PD. Coiled-coil-helix-coiled-coil-helix domain-containing 2 (CHCHD2) was identified as associated with autosomal dominant PD. However, the mechanism of CHCHD2 in PD remains unclear.Methods::Short hairpin RNA (ShRNA)-mediated CHCHD2 knockdown or lentivirus-mediated CHCHD2 overexpression was performed to investigate the impact of CHCHD2 on mitochondrial morphology and function in neuronal tumor cell lines represented with human neuroblastoma (SHSY5Y) and HeLa cells. Blue-native polyacrylamide gel electrophoresis (PAGE) and two-dimensional sodium dodecyl sulfate-PAGE analysis were used to illustrate the role of CHCHD2 in mitochondrial contact site and cristae organizing system (MICOS). Co-immunoprecipitation and immunoblotting were used to address the interaction between CHCHD2 and Mic10. Serotype injection of adeno-associated vector-mediated CHCHD2 and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration were used to examine the influence of CHCHD2 in vivo.Results::We found that the overexpression of CHCHD2 can protect against methyl-4-phenylpyridinium (MPP+)-induced mitochondrial dysfunction and inhibit the loss of dopaminergic neurons in the MPTP-induced mouse model. Furthermore, we identified that CHCHD2 interacted with Mic10, and overexpression of CHCHD2 can protect against MPP +-induced MICOS impairment, while knockdown of CHCHD2 impaired the stability of MICOS. Conclusion::This study indicated that CHCHD2 could interact with Mic10 and maintain the stability of the MICOS complex, which contributes to protecting mitochondrial function in PD.

Objective:To evaluate the effectiveness of staged modular theory and practice lectures in laparoscopic training.Methods:In this study, a large group of medical students and a large group of residents were selected, and each of the two groups was divided into an experimental group and a control group respectively. The experimental group received staged modular theory and practice lectures based on the conventional trainings, and the control group only underwent conventional laparoscopic skills training and instruction. Before and after the training, the completion time of 6 basic laparoscopic operations, laparoscopic suture time, suture quality and the number of accidental injuries were recorded and scored in each group according to the assessment criteria. SPSS 26.0 was used for statistical analysis of the assessment data. Independent samples t-test or Mann-Whitney U test was used for inter-group comparison between the two large groups, and paired samples t-test or Wilcoxon sign rank sum test was used for intra-group comparison before and after the training. Results:Compared with the control group, the experimental group of medical students that had undergone staged modular theory and practice lectures had a shorter time to complete basic laparoscopic operations, faster speed of suturing ( t=6.18, P<0.001) and higher quality of suturing ( t=4.17, P<0.001) and fewer accidental injuries ( Z=-2.03, P=0.043); the experimental group of residents that had undergone staged modular theoretical and practical lectures had a shorter time to complete basic laparoscopic operations, faster speed of suturing ( t=3.31, P=0.002) and higher quality of suturing ( t=3.68, P=0.001) and fewer accidental injuries ( Z=-2.44, P=0.015). Conclusion:The staged modular theory and practice lectures are able to further improve the quality of basic laparoscopic skills training.

Objective:To investigate the epidemiological characteristics of children respiratory pathogen infection in Ningbo city, and provide scientific reference for clinical prevention, control, diagnosis and treatment.Methods:A total of 3 103 specimens were collected from the children with acute respiratory infections in Ningbo Women and Children′s Hospital from July to October 2019. Thirteen common respiratory pathogens were detected by multiple reverse transcription-polymerase chain reaction (RT-PCR) based on capillary electrophoresis, and the result were analyzed.Results:Of the 3 103 specimens, 2 053 were positive for respiratory pathogens, with a total positive rate of 66.16%. The detection rates of Mycoplasma pneumoniae (Mp) 33.19% (1 030/3 103), human rhinovirus (HRV) 24.20% (751/3 103), and human adenovirus (HADV) 8.77% (272/3 103) were in the first three. The positive rates of pathogens in the male and female children were 65.70% (1 157/1 761) and 66.77% (896/1 342), respectively, and the difference was not statistically significant( χ2=0.386, P=0.535); the positive rates in the age groups of ≤1 year old, 1~≤3 years old, 3~≤6 years old and 6~≤16 years old were 58.02% (626/1 079), 74.37% (589/792), 73.78% (543/736) and 59.48% (295/496), respectively, and the difference among the groups was statistically significant( χ2=84.770, P=0.000); the positive rates in July to October were 71.64% (341/476), 68.81% (536/779), 67.01% (648/967) and 59.93% (528/881), respectively, the difference was statistically significant( χ2=24.395, P=0.000); the single infection rate was 52.30% (1 623/3 103), and the mixed infection rate of two or more pathogens was 13.86% (430/3 103), most of which were mixed infections of two pathogens. Conclusions:The main respiratory pathogens of children in Ningbo city were Mp and HRV, and the detection rates of respiratory pathogens varied among different ages and months.

Objective:To investigate the characteristics of senile neurodegenerative diseases (NDDs) inpatients in south China, especially in Guangdong province, and explore the influencing factors for their medical expenses.Methods:The medical records of 7231 patients with NDDs≥65 years were collected in the electronic health database of our hospital from January 2010 to December 2019, including gender, age, admission ways, chief complaints, length of hospital stays and medical expenses. On the basis of median of the medical expenses (21 345 yuan) of these patients, they were divided into low cost (<21 345 yuan) group and high cost (≥21 345 yuan) group. Univariate Logistic analysis and multivariate Logistic regression analysis were conducted to screen the influencing factors for medical expenses and the independent influencing factors.Results:(1) The main age group of geriatric inpatients with NDDs were 70-79 years (40.96%); the admission source was mainly outpatient (56.70%), and length of hospital stays of a large percent of patients (44.50%) were 8-14 d. (2) From 2010 to 2019, the number of hospitalized geriatric patients with NDDs showed an increasing trend year by year, the overall trend of length of hospital stays was shortened, and the medical expenses showed gradual increase; the causes of hospitalization, percentages of patients caused by infection, abnormal blood pressure and water-electrolyte metabolism disturbances showed decreased trend, percentages of patients caused by heart diseases, cerebrovascular accidents and mental-psychological diseases showed increased trend, and the proportions of patients caused by fracture/trauma/wound injuries were generally stable. The proportion of patients returning home and mortality rate after hospital discharge were declined, and the proportion of patients returning to other medical or community institutions was increased. (3) Living in ICU, length of hospital stays, diabetes, nosocomial infection, chronic kidney disease, urinary tract infection, tumble, body mass index, and anticholinergic drugs were independent risk factors influencing the medical expenses ( P<0.05). Conclusions:An aging trend is noted in patients with NDDs; the number of hospitalized patients and medical expenses increase year by year, and the length of hospital stays gradually decreases. In view of the many factors that influence the medical expenses of this disease, it is suggested to develop the corresponding standardized treatment plan for the main influencing factors in clinical practice.