Objective:To investigate the mechanism by which baicalein alleviates osteoarthritis by regulating Sirtuin (Sirt) to inhibit the senescence of fibroblast-like synoviocytes (FLS).Methods:FLS were extracted from synovium of non-osteoarthritis patients (3 patients) and osteoarthritis patients (3 patients), and assessed the senescence of FLS in osteoarthritis patients by Western blot, β-galactosidase staining and immunofluorescence, and examined the changing levels of Sirt family proteins and mRNAs in FLS. The relative expression level of Sirt1 in FLS was knocked down by transfection with Sirt1 siRNA, and the antioxidative capacity and senescence degree of FLS were analyzed. Chondrocytes were co-cultured with FLS and functional changes in chondrocytes were detected by Western blot. After inducing FLS senescence with bleomycin (BLM), different concentrations of baicalein were administered to detect the expression levels of Sirt1, p16, and p21 proteins and intracellular reactive oxygen species (ROS). The expression level of Sirt1 in FLS was knocked down, and BLM and baicalein treatments were applied. The relative expression levels of Sirt1 and ROS were detected using Western blot and ROS assays. After BLM-induced FLS senescence, baicalein and Compound C were added. The relative expression levels of phosphorylated AMP-activated protein kinase (pAMPK), nuclear factor erythroid 2-related factor 2 (NRF2), p16, p21, and ROS were detected by Western blot and ROS assays.Results:The relative expression of p16 and p21 in FLS in the osteoarthritic group were 3.66±0.38 and 3.55±0.34, which were higher than those in non-osteoarthritis group 1.00±0.07 and 1.00±0.09 ( t=11.860, P<0.001; t=12.520, P<0.001). The relative expression levels of Sirt1 and Sirt6 in FLS in the osteoarthritis group were 0.30±0.04 and 0.16±0.01, which were smaller than those in the non-osteoarthritis group 1.00 ± 0.03 and 1.00±0.04 ( t=23.840, P<0.001; t=34.130, P<0.001). After baicalein treatment, the relative expression of ROS was 2.58±0.28, 1.65±0.14 and 1.00±0.24 in the BLM, BLM+Bai and control groups, respectively, with statistically significant differences between the groups ( F=35.700, P<0.001), which was greater in the BLM and BLM+Bai than in the control group, and lower in the BLM+Bai than in the BLM group ( P<0.05). The relative expression of pAMPK and NRF2 was 0.28±0.02 and 0.38±0.09 after Compound C+Baicalein treatment, which was lower than that of 0.56±0.07 and 0.60±0.08 in the BLM+Bai group ( P<0.05). The relative expression of ROS increased from 1.75±0.16 to 3.45±0.12 ( P<0.001). The proportion of positivity in the BLM+Bai+Compound C, BLM+Bai and control groups was 47.30%±4.29%, 18.18%±3.89% and 7.70%±3.53% ( F=109.700, P<0.001), respectively, with the BLM+Bai+Compound C group being higher than that in the BLM+Bai group ( P<0.05). Conclusion:The downregulation of Sirt1 expression in patients with osteoarthritis leads to FLS senescence and accelerates the progression of osteoarthritis. Baicalein can inhibit FLS senescence by activating Sirt1/AMPK/NRF2 pathway, which may delay the progression of osteoarthritis and improve the function of chondrocytes.

Objective:To evaluate the relationship between the mechanism underlying the antidepressant effect of S-ketamine and hippocampal gamma-aminobutyric acid B receptor (GABA BR) in mice. Methods:A total of 54 male C57BL/6(B6) mice, aged 8 weeks, weighing 25-30 g, were used in this study. Forty mice were selected to develop the depression model by chronic social defeat stress. Twenty-six depression-susceptible mice were screened out by social avoidance test at day 11 after developing the model and divided into 2 groups ( n=13 each) by a random number table method: depression-susceptible group (Sus group) and depression-susceptible + S-ketamine group (Sus + S-ket group). The remaining 14 mice served as control group (C group). Starting from day 12 after developing the model, S-ketamine 10 mg/kg was intraperitoneally injected every day for 3 consecutive days in Sus+ S-ket group, while the equal volume of normal saline was given instead in C group and Sus group. The open field test was performed at 1 h after the last administration, and the total distance of movement was recorded. The forced swimming test was performed at 1 day after the open field test, and the immobile time was recorded. The sucrose preference test was performed to calculate the proportion of sucrose consumption at 1 day after the forced swimming test. One hour after the end of behavioral test, mice were sacrificed, and the hippocampal tissues were removed. Western blot was used to detect the expression of GABA BR1, GABA BR2, mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), brain-derived neurotrophic factor (BDNF), tyrosine kinase receptor B (TrkB), phosphorylated TrkB (p-TrkB), glutamate receptor 1 (GluR1) and postsynaptic dense protein 95 (PSD95). The p-mTOR/mTOR ratio and p-TrkB/TrkB ratio were calculated. The fluorescence intensity of BDNF in hippocampal CA1 region was detected by immunofluorescence. The number of dendritic spines in hippocampal CA1 region was measured by Golgi staining. Results:In the open field test, no statistically significant difference in the total distance was detected among the three groups ( P>0.05). Compared with C group, the immobile time in the forced swimming test was significantly prolonged, the proportion of sucrose consumption was decreased, the expression of hippocampal GABA BR1, GABA BR2, BDNF, GluR1 and PSD95 was down-regulated, and the ratios of p-mTOR/mTOR and p-TrkB/TrkB were decreased, the fluorescence intensity of BDNF and total number of dendritic spines in the hippocampal CA1 region were decreased in Sus group ( P<0.05), and no significant change was found in the parameters mentioned above in Sus+ S-ket group ( P>0.05). Compared with Sus group, the immobile time in the forced swimming test was significantly shortened, the proportion of sucrose consumption was increased, the expression of hippocampal GABA BR1, GABA BR2, BDNF, GluR1 and PSD95 was up-regulated, the ratios of p-mTOR/mTOR and p-TrkB/TrkB were increased, and the fluorescence intensity of BDNF and total number of dendritic spines in the hippocampal CA1 region were increased in Sus+ S-ket group ( P<0.05). Conclusions:The mechanism underlying the antidepressant effect of S-ketamine may be related to up-regulation of hippocampal GABA BR expression, activation of mTOR-BDNF signaling pathway, and improvement in synaptic plasticity in mice.

BACKGROUND: LY01005 (Goserelin acetate sustained-release microsphere injection) is a modified gonadotropin-releasing hormone (GnRH) agonist injected monthly. This phase III trial study aimed to evaluated the efficacy and safety of LY01005 in Chinese patients with prostate cancer. METHODS: We conducted a randomized controlled, open-label, non-inferiority trial across 49 sites in China. This study included 290 patients with prostate cancer who received either LY01005 or goserelin implants every 28 days for three injections. The primary efficacy endpoints were the percentage of patients with testosterone suppression ≤50 ng/dL at day 29 and the cumulative probability of testosterone ≤50 ng/dL from day 29 to 85. Non-inferiority was prespecified at a margin of -10%. Secondary endpoints included significant castration (≤20 ng/dL), testosterone surge within 72 h following repeated dosing, and changes in luteinizing hormone, follicle-stimulating hormone, and prostate specific antigen levels. RESULTS: On day 29, in the LY01005 and goserelin implant groups, testosterone concentrations fell below medical-castration levels in 99.3% (142/143) and 100% (140/140) of patients, respectively, with a difference of -0.7% (95% confidence interval [CI], -3.9% to 2.0%) between the two groups. The cumulative probabilities of maintaining castration from days 29 to 85 were 99.3% and 97.8%, respectively, with a between-group difference of 1.5% (95% CI, -1.3% to 4.4%). Both results met the criterion for non-inferiority. Secondary endpoints were similar between groups. Both treatments were well-tolerated. LY01005 was associated with fewer injection-site reactions than the goserelin implant (0% vs . 1.4% [2/145]). CONCLUSION: LY01005 is as effective as goserelin implants in reducing testosterone to castration levels, with a similar safety profile. TRIAL REGISTRATION ClinicalTrials.gov, NCT04563936.
Humans ; Male ; Antineoplastic Agents, Hormonal/therapeutic use* ; East Asian People ; Gonadotropin-Releasing Hormone/agonists* ; Goserelin/therapeutic use* ; Prostate-Specific Antigen ; Prostatic Neoplasms/drug therapy* ; Testosterone

Objective To discuss the etiology and clinical characteristics of human rhinovirus (HRV) as pathogen of central nervous system infection .Methods Two hundred and five cerebrospinal fluid (CSF) specimens were collected from children with fever and convulsions who were admitted to the pediatric intensive care unit with suspicion of central nervous system infection from June 2011 to December 2012 .Genome Lab Genetic Analysis System (GeXP) was applied to detect HRV from CSF .Specimens with positive results were amplified by nested reverse transcription‐polymerase chain reaction and followed by gene sequencing . Clinical data of HRV positive cases were analyzed . Results Of the 205 CSF specimens ,7 samples were positive for HRV ,which were composed of 2 HRV‐A ,1 HRV‐B and 4 HRV‐C (including 1 HRV‐Ca) .There were 6 boys and 1 girl among the 7 positive cases for HRV .Six children were less than 3 years old ,except one was 9 years old .The onset time was mainly concentrated between September and October . The main clinical manifestations were fever and convulsions . The clinical diagnosis before the pathogen confirmation included viral encephalitis ,epilepsy ,febrile convulsion ,benign infantile convulsions with mild gastroenteritis (CwG ) and hand‐foot‐and‐mouth disease ( HFMD ) . Although the disease severity of the 7 cases varied ,all ended with favorable prognosis .Conclusions HRV is one of pathogens of viral central nervous system infection .All types of HRV can cause central nervous system infection ,among which HRV‐C accounts for the majority .The clinical manifestations of HRV central nervous system infection could mimic febrile convulsion ,CwG and HFMD .

Objective To investigate familial hyperlipoproteinemia and the features of familial hyperlipoproteinemia in Lanzhou . Methods Data were from previous studies on the subject .Families of hyperlipoproteinemia were screening ,questionnaires were col-lected ,physical examination and laboratory data of family members were also colleted to analysis the characteristics of familial hy-perlipoproteinemia .Results A total of 39 familial hyperlipoproteinemia families were enrolled in the study ,including 280 family members .There were 15 core families ,11 single-parent families ,and 13 orphaned families .There were 6 familial hypercholesterol-emia families ,9 familial hypertriglycerides families ,24 mixed familial hyperlipidemia families .The children of the first generation ac-counted for 63 .2% of the total number of people enrolled in the study ,the father generation accounted for 14 .3% ,the children of the second geration accounted for 22 .5% .Conclusion In the survey ,the most common type of familial hyperlipidemia was mixed familial hyperlipidemia .The father generation was majority .The member of core families was less than incomplete families .

Objective To discuss the medium-term follow-up of clinically insignificant residual fragments (CIRF) after minimally invasive percutaneous nephrolithotomy lithotripsy (MPCNL).Methods The clinical data of 72 patients with CIRF medium-term follow-up were analyzed retrospectively.Results Seventy-two patients with CIRF.The anatomical distribution of CIRF was 10 at upper pole,15 at middle,35 at lower,10 at renal ureteropelvie junction and 2 at upper and lower pole.Stone analysis showed that 41 cases of calcium oxalate calculi,16 of calcium oxalate calculi mixed with carbonate calculi,3 calcium oxalate calculi mixed with uric acid,4 calcium oxalate calculi mixed with struvite stone,3 struvite stone,2 uric acid stone and 3 carbonate apatite mixed with struvite stone.Fifteen cases had clinical symptoms,including 2 renal colic pain,8 hematuria,5 lower urinary tract symptoms,4 cases CIRF located in upper pole,1 case in middle pole,4 cases in lower pole,6 cases in ureteropelvic junction,the incidence of clinical symptoms in ureteropelvic junction was significantly higher than that in other locations (6/10 vs.4/12,1/15,4/37,P ＜0.05).Eight cases required surgical procedure,5 cases underwent extracorporeal shock wave lithotripsy,3 cases with ureteral CIRF were performed with ureteroscopic lithotripsy.CIRF were clear after surgery,7 patients with ureteral CIRF had renal colic pains.The stones were excluded after spasmolytic analgesic treatments.Conclusions CIRF can be located variously in the kidney and ureter.Most CIRF are calcium oxalate calculi and locate in the lower pole.Patients with the history of previous open surgery or extracorporeal shock wave lithotripsy are more likely to get CIRF.Medium-term follow-up of CIRF reveals that CIRF located in the renal ureteropelvis junction are more likely to have clinical symptoms.

Objective To explore the status and risk factors of the dyslipidemia among health examination population of Lanzhou for providing the intervention measures .Methods According to the stratified cluster random sampling method ,4 505 health exami-nation individuals were recruited for the study from 5 hospitals in the Lanzhou region through questionnaire ,biochemical analysis . Results Prevalence of dyslipidemia of the population was 45 .79% ,high TG was the main type .The level of serum TC ,TG ,HDL-C and LDL-C were (5 .27 ± 1 .08) ,(1 .74 ± 1 .38) ,(1 .41 ± 0 .43) and (2 .83 ± 0 .82)mmol/L .The prevalence was 53 .49% in male , and 34 .93% in female .The prevalence was higher among the group of 35- <45 years old for male and 55- <65 years old for fe-male .The level of HDL-C was low among young people .There was aggregation of risk factors among the participants with dyslipi-demia .Non-conditional logistic regression analysis showed that the risk factors were age (OR=1 .701) ,overweight (OR=5 .560) , abdominal obesity(OR=2 .398) ,smoking(OR=0 .545) ,intake of greasy diet(OR=5 .313) ,sleep quality(OR=2 .005) and diastolic blood pressure(OR=3 .061) .Conclusion Lipid disorders becomes a serious problem in the health examination population ,measures such as rational diet ,weight control ,sleep improvement ,pressure control and quiting smoking must be taken .

Objective To determine the incidence of avascular necrosis after closed reduction and cannulated compression screws fixation of intra-capsular femoral neck fracture and to investigate the multiple factors correlated to avascular femoral head necrosis.Methods All the patients of intra-capsular femoral neck fracture who accepted closed reduction and cannulated compression screws fixation between 2001 and 2010 in Jiangsu Province were reviewed in multi-centers retrospectively.The multiple factors were analyzed including age,gender,affected side,mechanism of injury,fracture classification,procedure delay,quality of reduction,time of full-weight-bearing,configuration of the screws and removal of the screws.Multiple correlation factors were analyzed with SPSS 13.0 statistic system.Results Complete case records were documented in 1849 cases who were followed up for an average 6.5±2.7 years (range,2-10 years).Avascular necrosis occurred in 246 cases (13.3％).The average time of diagnosis of avascular necrosis was 17±4.6months (range,8-72 months) after injury.The average Harris score of the hips which didn't develop to avascular necrosis was 93.8±8.9 (range,78～100) at the last follow up.Multiple correlation factors analysis indicated that displacement degrees of fracture and the quality of reduction were significantly correlated to the incidence of avascular necrosis (OR=2.078,3.423).Conclusion Closed reduction and cannulated compression screws fixation after the intra-capsular femoral neck fracture can get satisfactory results.Displacement degrees of fracture and the quality of reduction are significantly correlated to the incidence of avascular femoral head necrosis.

Objective To evaluate the clinical effects and safety of percutaneous nephrolithotomy (PCNL) by middle renal calice used as the main target for the treatment of staghorn stones with the combination of pneumatic and ultrasonic lithotrite.Methods Clinical data of 73 patients underwent PCNL by middle renal calices as main access with 57 incomplete staghorn stones and 35 complete staghorn stones.To observe the situation calculus removal rate and complications.Results Seventy cases (88 sides) underwent one session PCNL by single access tract (middle caliees),3 cases (4 sides) underwent one session PCNL by double access tracts (2 cases by middle and low calices,1 case by up and middle caliees).After the first period of lithoclasty,17 patients (25 sides) residual stones and the stone removal rate 72.8％ (67/92),among these patients,1 case (1 side) had fragments of lateral renal calyeeal stones with no further treatment.Other 16 cases (24 sides)underwent second session PCNL,all were treated by single access tract (middle calices) and 2 cases (2 sides)had extracorporeal shock wave lithotripsy before the second PCNL.After the second period of lithoclasty,76 sides composed of 27 complete staghorn stones and 49 incomplete staghorn stones had no residual fragments with the stone removal rate 82.6％ (76/92).The operative time lasted 120-320 min.Hemoglobin dropped 1-4 g/L,11 cases in the operation procedure and 3 cases after operation needed blood transfusion respectively.One case of renal pelvic infection after operation and 1 case had split renal dysfunction with peri-parenchyma infection.The hospitalization time was 9-18 days.Conclusion It is effective and safe to perform PCNL for staghorn stone by middle calices as a main access.Combining pneumatic and ultrasonic lithotrite will be very useful with high stone clearance,short procedure time and less complications.

4-Hydroxybutyric acid (4HB) is a psychotropic drug used for polymer synthesis such as poly (4-hydroxybutyric acid) (P4HB) and poly (3-hydroxybutyric acid-co-4-hydroxybutyric acid) (P3HB-co-4HB). 1,4-butanediol (BD) can be converted to 4-hydroxybutyric acid by alcohol dehydrogenase (DhaT) and aldehyde dehydrogenase (AldD). In this study, high efficiency promoters including T7 promoter and P(Re) promoter were cloned to increase expression of dhaT and aldD, and thus accelerate the conversion from BD to 4HB. A. hydrophila 4AK4 (pZQ01), the recombinant strain under the control of T7 promoter, produced 6.00 g/L 4HB from 10 g/L BD with the productivity increased by 43.20%. While A. hydrophila 4AK4 (pZQ04), the strain under the control of T7 promoter, produced 4.87 g/L 4HB from 10 g/L BD, and the productivity was increased by 16.23%. Thus, the gene expression was increased by T7 and P(Re) promoters, leading to an accelerated biosynthesis of 4HB.
Aeromonas hydrophila ; genetics ; metabolism ; Genetic Engineering ; Hydroxybutyrates ; metabolism ; Promoter Regions, Genetic ; genetics ; Recombination, Genetic