Objective To isolate and identify the potential binding partners of LRRK2,a gene linked to both dominant familial form and sporadic form of Parkinson's disease,thus to further our knowledge of its function.Methods We used a sequence containing full-length of COR domain and part of ROC and MAPKKK domain as bait.The bait amplified by polymerase chain reaction(PCR) was then cloned into a yeast expression plasmid pGBKT7.After being sequenced and analyzed,pGBKT7-bait was transformed into the yeast strain AH109.Western blot was performed to confirm the expression of pGBKT7-bait in AH109 yeast strain.Then human fetal brain cDNA library was trarnsformed into that yeast strain.which could express pGBKT7-bait fusion protein.The yeast strain which contained pGBKT7-bait and human fetal brain cDNA library was plated on quadruple dropout medium (SD/-Trp/-Leu/-His/-Ade)containing X-a-gal.We retested these positive colonies using 2 independent yeast strains AH109 contained pGBKT7-bait or pGBKT7,respectively.At last,these plasmids isolated from these true positive colonies were analyzed by bioinformatics.Results We obtained 9 true positive colonies,these colonies were sequenced, and we performed sequence Blast in GenBank.Three colonies of the 9 positive colonies were not in open reading-frames.Among other 6 colonies,there were known proteins including spermatid perinuclear RNA-binding protein(STRBP)and BCL2-associated athanogene 5 isoform b(BAG5),as well as unknown proteins including tyrosine phosphatase non-receptor type(PTPN23),1(3)mbt-like 3 isoform b(L3 MBTL3),RALY RNA binding protein-like isoform 1(RALYL),and Homo sapiens mRNA for KIAA1783 protein,partial cds(KIAA 1783).Conclusion True positive colonies of LRRK2 are successfully obtained by the yeast 2-hybrid.Our screened proteins may provide a new research clue for revealing biological functions of LRRK2,pathogenesis of Parkinson's disease,and other neurodegerations.

Objective:To detect the expression of MYBL2 gene in gastric adenocarcinoma tissue and its effects on cell proliferation and invasion. Methods:A total of 100 cases of gastric adenocarcinoma tissue and 100 cases of paracancerous tissue were selected from patients who received surgery in The People's Hospital of Yuhuan between January 2017 and December 2020. Gastric adenocarcinoma cell lines MGC-803 were transfected with MYBL2 siRNA and siRNA control. The cells not transfected were used as controls. MYBL2 gene expression in gastric adenocarcinoma tissue and paracancerous tissue as well as MGC-803 were determined by quantitative real time-polymerase chain reaction. MGC-803 cell proliferation was determined by MTT. The invasive ability of MGC-803 cells was determined by Transwell assay. The migration ability of MGC-803 cells was determined by Scratch testing. MYBL2 protein expression in gastric adenocarcinoma tissue and paracancerous tissue as well as MGC-803 cells was determined by western blotting. Results:The relative mRNA expression of MYBL2 in gastric adenocarcinoma tissue was significantly higher than that in paracancerous tissue [(0.65 ± 0.17) vs. (0.18 ± 0.05), t = 26.52, P < 0.05). The relative mRNA expression of MYBL2 in the MYBL2 siRNA group (0.29 ± 0.07) was significantly lower than that in the control group (0.73 ± 0.12) and siRNA group (0.71 ± 0.16, t = 5.48, 4.16, both P < 0.05). MTT assay showed that after 24 and 48 hours of culture, MGC-803 cell proliferation rate in the MYBL2 siRNA group [(40.95 ± 5.46)%, (52.12 ± 12.27)%] was significantly lower than that in the control group [(67.84 ± 6.45)%, (87.83 ± 9.96)%] and siRNA group [(66.98 ± 7.85)%, (85.98 ± 10.24)%, t = 5.51, 3.91, 4.71, 3.67, all P < 0.05]. MGC-803 cell invasion rate in the MYBL2 siRNA group [ (62.12 ± 6.43)%] was significantly lower than that in the control group [(89.74 ± 6.56)%] and siRNA group [(88.83 ± 7.85)%, t = 5.20, 4.55, both P < 0.05]. The number of MGC-803 cells migrated in the MYBL2 siRNA group [(4.32 ± 0.84) × 10 3] was significantly lower than that in the control group [(8.95 ± 1.64) × 10 3] and siRNA group [(8.83 ± 1.78) × 10 3, t = 4.35, 3.96, both P < 0.05]. The gray value of MYBL2 protein in the gastric adenocarcinoma tissue was (0.56 ± 0.15), which was significantly higher than that in the paracancerous tissue [(0.23 ± 0.07), t = 19.93, P < 0.001]. The gray value of MYBL2 protein in the MYBL2 siRNA group was (0.21 ± 0.03), which was significantly lower than that in the control group (0.67 ± 0.15) and siRNA group (0.65 ± 0.19) ( t = 5.20, 3.96, both P < 0.05). Conclusion:MYBL2 gene is highly expressed in gastric adenocarcinoma tissue. siRNA silencing MYBL2 can decrease the ability of MGC-803 cells to proliferate, invade and migrate and downregulate MYBL2 expression. This study is highly innovative and scientific.

Objective To analyze the factors influencing the early recovery of renal function in patients with sepsis-associated acute kidney injury(SA-AKI).Methods A retrospective analysis was conducted on 86 SA-AKI patients treated in the Intensive Care Unit at Zhongshan Hospital,Fudan University from January 2021 to December 2022,who met both the Sepsis 3.0 diagnostic criteria and the AKI diagnostic standards.Patients were divided into a recovery group and a non-recovery group based on whether their renal function recovered within 7 days after AKI onset.Clinical data and laboratory tests of patients were compared between the two groups.Univariate and multivariate logistic analyses were used to identify risk factors affecting renal function recovery in SA-AKI patients,and ROC curve was utilized to evaluate the predictive value of these factors for early renal function recovery in SA-AKI patients.Results The renal function of 37(43.02％)patients recoveried.Compared with the recovery group,the renal replacement therapy rate,in-hospital mortality and 28-day mortality of patients in the non-recovery group were higher(P＜0.001).The multivariate logistic analysis showed that age,APACHE Ⅱ score,urine output,urine neutrophil gelatinase-associated lipocalin(NGAL),and norepinephrine dose were independent related factors affecting renal function recovery in SA-AKI patients(P＜0.05).The final model logit(P)=-4.091+0.001×urine NGAL-0.001 Xurine volume+0.040 ×age+0.073 × APACHE Ⅱ score+1.906 × norepinephrine dose.The AUC of model predicting early SA-AKI recovery was 0.823,with 73.5％of sensitivity,and 81.1％of specificity.Conclusions In SA-AKI patients,age,APACHE Ⅱ score,urine output,urine NGAL,and the dose of norepinephrine independently affect early renal function recovery,and the combined assessment of these indicators has predictive value for the early renal recovery in these patients.

Objective? To explore the effect of multi-mode interactive continuous nursing on home rehabilitation and complications of patients after total hip arthroplasty(THA). Methods? From January 2017 to July 2018, patients undergoing THA in Zhejiang Provincial People's Hospital were selected by purposive sampling method. They were randomly divided into two groups according to visit numbers. There were 43 effective cases in the control group and 42 in the observation group. The control group adopted the traditional telephone follow-up mode, while the observation group adopted the multi-mode interactive continuous nursing intervention. The Modified Barthel Index(MBI), Harris Hip Score(HHS) and 6-Minute Walking Tes(t 6MWT) were used to compare the rehabilitation effects of the two groups at discharge, 1, 3 and 6 months after operation, and the complications, unplanned hospital visits and hospitalizations of the two groups were compared. Results? There was no significant difference in each index between the two groups at discharge (P＞ 0.05). The scores of MBI in the observation group were (78.57±7.43), (90.36±7.76), (94.41±7.51) respectively at 1, 3 and 6 months after operation, which were higher than those in the control group [(70.35±7.60), (82.40±7.82), (91.86±8.80)], and there were significant differences between the two groups at 1 and 3 months after operation (P＜ 0.05). The scores of HHS in the observation group were (82.05±7.40), (87.52±6.94), (91.17±7.40) respectively, higher than those in the control group (78.40±7.51), (84.09±7.23), (88.37±8.46), there were significant differences between the two groups at 1 and 3 months after operation (P＜ 0.05). The distance of 6MWT in the observation group were (174.76±35.04)m, (241.55±40.67)m, (282.86±46.46) m, higher than those in the control group (151.61±42.52)m, (214.42±48.59)m, (260.35±48.05) m. There were significantly statistical differences in the 1, 3 and 6 months after operation (P＜0.05). In the observation group, there were 1 case of fracture, 9 cases of unplanned hospital visit or hospitalization, 2 cases of dislocation, 1 case of infection and 20 cases of unplanned hospital visit or hospitalization in the control group. Conclusions? Continuous nursing with multi-mode interaction can improve the effect of home rehabilitation after THA, effectively improve the early and long-term hip function of patients, prevent and reduce complications, and improve the quality of life of patients.

BACKGROUND: Osteoporosis (OP) has become a major public health issue, threatening the bone health of middle-aged and elderly people from all around the world. Changes in the gut microbiota (GM) are correlated with the maintenance of bone mass and bone quality. However, research results in this field remain highly controversial, and no systematic review or meta-analysis of the relationship between GM and OP has been conducted. This paper addresses this shortcoming, focusing on the difference in the GM abundance between OP patients and healthy controls based on previous 16S ribosomal RNA (rRNA) gene sequencing results, in order to provide new clinical reference information for future customized prevention and treatment options of OP. METHODS: According to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA), we comprehensively searched the databases of PubMed, Web of Science, Embase, Cochrane Library, and China National Knowledge Infrastructure (CNKI). In addition, we applied the R programming language version 4.0.3 and Stata 15.1 software for data analysis. We also implemented the Newcastle-Ottawa Scale (NOS), funnel plot analysis, sensitivity analysis, Egger's test, and Begg's test to assess the risk of bias. RESULTS: This research ultimately considered 12 studies, which included the fecal GM data of 2033 people (604 with OP and 1429 healthy controls). In the included research papers, it was observed that the relative abundance of Lactobacillus and Ruminococcus increased in the OP group, while the relative abundance for Bacteroides of Bacteroidetes increased (except for Ireland). Meanwhile, Firmicutes, Blautia, Alistipes, Megamonas, and Anaerostipes showed reduced relative abundance in Chinese studies. In the linear discriminant analysis Effect Size (LEfSe) analysis, certain bacteria showed statistically significant results consistently across different studies. CONCLUSIONS: This observational meta-analysis revealed that changes in the GM were correlated with OP, and variations in some advantageous GM might involve regional differences.
Middle Aged ; Aged ; Humans ; Gastrointestinal Microbiome/genetics* ; RNA, Ribosomal, 16S/genetics* ; Genes, rRNA ; Osteoporosis ; Feces