Objective: To investigate the impact of RHAG 808A variant, commonly identified in the Chinese population, on RhAG protein, RhD and RhCE antigens expression through in vivo and in vitro expression analysis. Methods: A missense mutation of RHAG gene (c. 808G>A, p. Val270Ile) with high frequency was found in KMxD database. Bioinformatics analysis was performed using Polyphen-2 and Provean software. High resolution melting (HRM) method was utilized to screen for the variant carriers in the blood donors. The expression of RhAG protein, RhD and RhCE antigens on the surface of red cells of variant carriers were detected via flow cytometry. Wild-type and mutant vectors of RHAG were constructed and transfected into HEK 293T cells for in vitro expression analysis. Then, the expression of RhAG protein, RhD and RhCE antigens were analyzed by flow cytometry. Results: Polyphen-2 and Provean software suggested that the amino acid change (p. Val270Ile) of RhAG protein may be harmful or neutral respectively. Among the 999 blood donors from Guangzhou Blood Center, 4 homozygous carriers and 99 heterozygous carriers of RHAG 808A mutant allele were identified. The frequency of this allele was 5.4% (107/1 998). No significant differences in RhAG protein, RhD and RhCE antigens expression level was identified between the homozygous carriers, heterozygous carriers of RHAG 808A variant allele and the wild-type individuals. In vitro analysis for antigen expression study obtained the similar results. Conclusion: The RHAG 808A variant allele commonly identified in the Chinese population has no effect on the expression of RhAG protein, RhD and RhCE antigens, so the variant should be a population polymorphism site.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

【Objective】 To study the effect of RHAG variants identified in Chinese population on mRNA splicing by minigene splicing assay(MSA) in vitro. 【Methods】 The pSplicePOLR2G minigene expression plasmids were constructed for 10 RHAG mutations with relatively high distribution frequency in Chinese population near splicing sites or synonymous mutations by analyzing the RHAG gene data in the KMxD database. Then, the wild-type and mutant plasmids were transfected into HEK 293T cells, and RNA was extracted 48 hours after transfection. After reverse transcription, specific primers were used for PCR amplification, and then agarose gel electrophoresis and capillary electrophoresis were performed to determine whether the mutations will affect the normal splicing of exons. 【Results】 MSA in vitro showed that 2 mutations (c.158-5delT, c. 807+ 3A>C) near the splicing site reduced the amount of normal transcripts slightly. The remaining 8 synonymous mutations(c.312G>A, c. 341+ 3G>A, c. 609C>T, c. 681G>A, c. 861G>A, c. 957T>A, c. 984T>C and c. 1139-7G>A) had no impact on the splicing of RHAG mRNA. 【Conclusion】 This study showed that RHAG gene was conservative in terms of splicing, and the mutations near splicing sites and synonymous mutations were less likely to cause abnormal splicing of RHAG gene.

The association among plasma trimethylamine-N-oxide (TMAO), FMO3 polymorphisms, and chronic heart failure (CHF) remains to be elucidated. TMAO is a microbiota-dependent metabolite from dietary choline and carnitine. A prospective study was performed including 955 consecutively diagnosed CHF patients with reduced ejection fraction, with the longest follow-up of 7 years. The concentrations of plasma TMAO and its precursors, namely, choline and carnitine, were determined by liquid chromatography-mass spectrometry, and the FMO3 E158K polymorphisms (rs2266782) were genotyped. The top tertile of plasma TMAO was associated with a significant increment in hazard ratio (HR) for the composite outcome of cardiovascular death or heart transplantation (HR = 1.47, 95% CI = 1.13-1.91, P = 0.004) compared with the lowest tertile. After adjustments of the potential confounders, higher TMAO could still be used to predict the risk of the primary endpoint (adjusted HR = 1.33, 95% CI = 1.01-1.74, P = 0.039). This result was also obtained after further adjustment for carnitine (adjusted HR = 1.33, 95% CI = 1.01-1.74, P = 0.039). The FMO3 rs2266782 polymorphism was associated with the plasma TMAO concentrations in our cohort, and lower TMAO levels were found in the AA-genotype. Thus, higher plasma TMAO levels indicated increased risk of the composite outcome of cardiovascular death or heart transplantation independent of potential confounders, and the FMO3 AA-genotype in rs2266782 was related to lower plasma TMAO levels.
Carnitine ; Choline/metabolism* ; Chronic Disease ; Heart Failure/genetics* ; Humans ; Methylamines ; Oxygenases ; Prospective Studies

OBJECTIVE: To analyze the action of miRNA-326 on its target gene BCL-XL and the molecular mechanism of platelet apoptosis regulated by miRNAs. METHODS: Dual-luciferase vectors containing respectively the wild-type and mutant 3'-untranslated region (3'UTR) fragments of the BCL-XL gene were constructed with firefly and renilla luciferases and transfected into 293T cells. Relative fluorescence intensities of the transfected cells were measured. RESULTS: Dual-luciferase reporter gene vectors for PsiCHECK- BCL-XL -3'UTR-WT (wild-type) and PsiCHECK- BCL-XL -3' UTR-MT (variant) were respectively constructed. Relative fluorescence intensities of the 293T cells co-transfected by miRNA-326 and PsiCHECK- BCL-XL -3'UTR-WT plasmid were significantly lower compared with the control group (co-transfected by a miRNA-326 negative sequence and PsiCHECK- BCL-XL -3' UTR-WT plasmid) ( P = 0.034). The relative fluorescence intensity was also significantly reduced in cells co-transfected by miRNA-326 and PsiCHECK- BCL-XL -3' UTR-WT plasmid compared with the mutant control group co-transfected by miRNA-326 and PsiCHECK- BCL-XL -3'UTR-MT plasmid (P = 0.022). CONCLUSION miRNA-326 may participate in the regulation of platelet apoptosis by acting on the 3'-UTR of the BCL-XL gene.

Objective: To investigate the joint effect of abdominal obesity in childhood and adulthood on adult hypertension. Methods: Based on the data from the China Health and Nutrition Survey (CHNS, 1993-2011), a total of 1 431 subjects who were investigated in both childhood (6 to17 years old) and young adulthood (18 to 35 years old) were selected for the analysis. According to waist circumference (WC) status (normal WC or abdominal obesity) in childhood and adulthood, all subjects were categorized into 4 groups. The multinomial logistic regression model was used to analyze the joint effect of abdominal obesity in childhood and adulthood on adult hypertension and pre-hypertension. Results: Compared to the subjects (n=1 057) who had normal WC in both childhood and adulthood, subjects with abdominal obesity in childhood but with normal WC (n=45) in adulthood didn′t have significantly increased risk of hypertension (OR=1.52, 95%CI: 0.19-12.06). In contrast, those who had normal WC in childhood and abdominal obesity in adulthood (n=289) had increased risk of hypertension (OR=6.48, 95%CI: 3.60-11.66). In addition, subjects with persistent abdominal obesity from childhood to adulthood (n=40) had the highest risk of hypertension (OR=15.98, 95%CI: 5.39-47.35). There was a similar trend for the association of abdominal obesity in childhood and adulthood with adult pre-hypertension, with the corresponding OR (95%CI) of 1.28 (0.66-2.49), 2.90 (2.17-3.89) and 3.49 (1.65-7.40), respectively. Conclusion The joint effect of abdominal obesity in childhood and adulthood increased the risk of adult hypertension and pre-hypertension. There was no statistical significance for subjects who had abdominal obesity in childhood but had normal WC in adulthood when compared to those with normal WC in both childhood and adulthood.

Abstract]? Objective? To investigate the relevant knowledge, attitude and practice (KAP) of the parents of children with cleft lip and palate, so as to provide reference for the further development of health education. Methods? From August 2016 to February 2017, parents of children with cleft lip and palate admitted in a ClassⅢ Grade A general hospital in Shanghai were selected in the study. General Information Questionnaire, General Self-efficacy Scale (GSES), Parents' Knowledge-Attitude-Practice Questionnaire were used in the survey and the status of their KAP on the disease as well as the influencing factors were analyzed. A total of 142 questionnaires were distributed with 116 valid ones collected, yielding an effective recovery rate of 81.7%. Results? 116 parents of children with cleft lip and palate scored (30.84±7.22),(34.35±5.96) and (29.42±5.74) respectively on disease-related KAP. Among them, "knowledge and attitude" scored a medium level, and 57.8% of the children's parents scored a low level on "practice". Univariate analysis showed that children's gender, mother's educational level, mother's smoking and family monthly income were the influencing factors of parents' knowledge about disease;mother's age and drinking were the influencing factors of parents' attitude;monther's age and smoking were the influencing factors of parents' practice (P＜ 0.05). There was a positive correlation between the scores of disease-related practice of parents and self-efficacy (r=0.211,P＜0.05). Conclusions? Parents of children with cleft lip and palate lack knowledge of cleft lip and palate disease, and have positive attitude but poor action. Through effective and individualized health education of cleft lip and palate, it is hopeful to change their understanding and practice about the disease and further promote the health of children with cleft lip and palate.

Objective To assess the clinical effect of Percutaneous transforaminal endoscopic spinal surgery for central lumbar spinal stenosis.Methods Total 92 patients (44 males and 48 females with average age of 68.6± 12.4 years) with central lumbar spinal stenosis were enrolled in our study during February 2012 and July 2016.All patients were treated with Percutaneous transforaminal endoscopic spinal surgery.Clinical and Radiographic evaluation were investigated on 1 week,3 months,6 months,12 months postoperatively and final follow-up.Visual analogue scale (VAS) for low back pain and leg pain,Oswestry disability index(ODI) for low back pain were used to evaluate the clinical efficacy of surgery.Pre and postoperative Dural sac cross sectional area(DSCA) was measured.And the correlation between changes of DSCA and clinical efficacy was analyzed.Results The operation time was 45-1 15 min.The mean operation time was 75±15 min.the mean blood loss was 15±5 ml (range 8-50 ml).All patients were followed for 12-46 months (24.5±5.3 months).VAS for low back pain,VAS for leg pain,and OD1 were significantly improved from 6.75± 1.28,7.79± 1.15 and 39.82％ ±5.06％ preoperatively to 2.21± 1.08,2.16± 1.14 and 9.82％ ±3.69％ at the latest follow-up.Dural sac cross sectional area significantly increased from 55.35±12.18 mm2 preoperatively to (102.36±15.38) mm2 at the latest follow-up.Correlation coefficient with DSCA change was-0.480 for ODI change,-0.612 for VAS(low back pain) change,-0.637 for VAS (leg pain)(P ＜ 0.05);obvious positive correlation existed between the change of DSCA and patient's clinical efficacy.It showed that the change of DSCA was positively correlated with the patient's clinical efficacy.The clinical results were excellent in 56 cases.good in 29,fair in 4,and poor in 3 based on the MacNab criteria.92.39％ demonstrated a good-to-excellent outcome.One case occurred cauterizing syndrome,3 cases of recurrence,and 2 cases of Dural tear.There was no severe vascular or nerve injury.Conclusion Percutaneous transforaminal endoscopic spinal surgery provides a new minimally invasive treatment for central lumbar spinal stenosis.It is safe and effective.It is found that the change of DSCA is positively correlated with the clinical efficacy of surgery.

Objective To evaluate the feasibility of needle-based confocal laser endomicroscopy (nCLE) for imaging of intra-abdominal tissues and organs in rabbit models in vivo. Methods The nCLE miniprobe was inserted through the 19-gauge needle into various intra-abdominal tissues and organs[omentum majus, liver, pancreas and psoas major (skeletal muscle)]. The nCLE images were acquired and real-time sequences of respective locations were recorded. Finally, nCLE image characteristics were compared with histopathologic findings. Results nCLE was successfully performed in intra-abdominal tissues and organs of five rabbit models. The microscopic structures of cells, glands and microvessels in the omentum majus, liver, pancreas and psoas major ( skeletal muscle) were visualized clearly, respectively. Characteristics of various intra-abdominal tissues and organs were displayed on nCLE images, which were correlated well with histological findings. Conclusion Imaging of intra-abdominal tissues and organs with nCLE in vivo is feasible in future clinical practice.

ABSTRACT:Objective To assess the effect of prolonged laparoscopic surgery on peritoneal mesothelial cells and fibrinolysis in humans.Methods We examined prospectively 1 6 consecutive patients who underwent laparoscopic surgery (LAP)and 2 1 patients who underwent conventional open surgery (OP)for high-medium rectal cancer with curative intent.During the procedure,biopsy of the parietal peritoneum was made before operation and at 45 min,90 min,and 120 min after operation.The tissue-type plasminogen activator (tPA)and plasminogen activator inhibitor-1 (PAI-1 )were determined by enzyme-linked immunosorbent assay in peritoneal tissues.The cellular injury was detected by LDH assay.The proliferation was quantified by MTT assay.Results PAI-1 activity in the peritoneal tissue was significantly lower in LAP group than in the OP group.tPA activity decreased after 45min of open surgery,but there was no significant change in the LAP group.With time extension,the LDH activity increased and the proliferation of the mesothelial cells decreased.Conclusion Preservation of a prolonged hypofibrinolytic state by inhibition of PAI-1 up-regulation during LAP may predispose patients to less postoperative peritoneal adhesion. The cellular injury becomes apparent and the proliferation is inhibited during prolonged laparoscopic surgery.