Objective To investigate the effects of celastrol on the cell growth and apoptosis of human gallbladder cancer NOZ cells,and explore its potential molecular mechanism.Methods NOZ cell were cultured in vitro.And CCK-8 assay,Annexin V-FITC/PI staining method,cell cycle analysis were conducted to investigate the effects of celastrol on the growth and apoptosis of NOZ cells after being treated with drugs.The mitochondrial membrane potential and Bax and Bcl-2 protein expression level were determined by Rhodamine 123 and Western blot,respectively.Results Celastrol could inhibit NOZ cell growth,and the IC50 value was 5.3 μmol/L.Annexin-V/PI staining showed that cell apoptosis of NOZ cells were induced as the celastrol concentration increased,and the apoptosis ratio of control group was 4.4％,while the apoptosis rates of the test groups (2,5,10 p mol/L) were 7.4％,27.1％ and 43.4％,respectively.In addition,cell cycle analysis revealed that celastrol could induce G1-phase arrest.The G1-phase rate of control group was 25.6％,while the G1-phase rates of the test groups (2,5,10 μmol/L) were 36.5％,45.7％ and 92.5％,respectively.The mitochondrial membrane potential was measured after treatment with celastrol and the results indicated that the mitochondrial membrane potential was significantly decreased.Western Blot showed that the protein expression of Bax increased and Bcl-2 decreased in a time-dependent manner after treatment with celastrol.Conclusions Celastrol may inhibit cell proliferation of human gallbladder cancer NOZ cells and induce cell apoptosis partly by inducing the loss of mitochondrial membrane potential.

Objective To study the clinical value of wrist magnetic resonance imaging (MRI) combined anti-cyclic citrullinated peptide antibody detection in the diagnosis of early rheumatoid arthritis (RA).Methods Forty five patients with early RA were selected as RA group,45 cases of patients without rheumatoid arthritis as non-RA group,and 43 cases of people with normal examination as control group.All subjects were given wrist MRI and anti-cyclic citrullinated peptide (CCP) antibody with the enzyme linked immunosorbent assay (ELISA).At the same time,clinical symptoms,physical signs,MRI manifestations,and laboratory indicators were collected.All results were statistically analyzed.Results Positive rate of MRI lesions and serum anti-CCP antibody in RA group were significantly higher than non-RA group and control group (P ＜0.05).The sensitivity and specificity of MRI (or anti-CCP antibody) for early RA were 88.88％ and 82.22％ (or 68.88％ and 91.11％).The sensitivity (64.44％) of MRI combined with anti-CCP antibody was decreased compared to individual; however,the specificity (100％) of MRI combined with anti-CCP antibody was increased.The correlation of MRI synovial scores and anti-CCP antibody levels was positively correlated (rs =0.612,P ＜ 0.05).MRI abnormal signs and joint disease activity score (DAS28) were positively correlated (rs =0.521,P ＜ 0.05).Anti-CCP antibody levels and DAS28 were positively correlated (rs =0.541,P ＜ 0.05).Conclusions MRI examination and combined with anti-CCP antibody detection is helpful to improve the diagnosis of early RA,and it provides a detection basis for dynamic assessment of RA condition changes.

Objective To master the area scope and epidemic situation of Kaschin-Beck disease (KBD) in Harbin,and to accumulate data and information for the midterm examination and evaluation of the 12th Five-Year Plan of the National Endemic Disease Prevention and Control.Methods In the original 10 counties (districts,cities) of Harbin with KBD,2 surveys were carried out from 2011 to 2015,5 townships were extracted in each county (district,city),3 villages were extracted in each township,clinical investigation and X-ray examination of 7-12 years old children were carried out;according to the criteria for Diagnosis of Kaschin-Beck Disease,a clinical survey was conducted in 2013 in the 10 counties (districts,cities) with KBD.According to the KBD prevalence and X-ray examination results in each county (district,city),reference to Criteria for Control of Kaschin-Beck Disease Areas,Criteria for Delimitation and Classification of Kaschin-Beck Disease Area,Evaluation Methods for Endemic Diseases Control and Elimination,the types of the disease areas,control and eliminating situation were determined;organization and management of the control and prevention measures were evaluated.Results Currently,there were 10 counties (districts,cities),373 villages,6 969 patients with KBD in Harbin.According to the KBD area division and control standard,369 villages were light KBD areas (prevalence of clinical degrees Ⅰ and above patients or detection rate of 7-12 children by X-ray for local residents ≤ 10％) and 4 villages were medium KBD areas (prevalence of clinical degrees Ⅰ and above patients or detection rate of 7-12 children by Xray for local residents ＞ 10％-≤20％).Clinical census showed that the people under 20 years had no cases with degree Ⅰ and above in each village,X-ray detection rate was 0 in 7-12 years old children,in line with the standards of history KBD areas.There were no clinical KBD cases in 7-12 years old children in all villages of KBD areas,positive detection rate of children by X-ray was 0 in the sampling KBD area villages,in line with the standards of eliminate technical standards of KBD area village.The management index scores of KBD were from 85 to 95 points in every area county (district,city),getting to the eliminating requirement which was more than 85 points.Conclusion The KBD areas have narrowed down and prevalence has declined significantly in Harbin,which has reached the area eliminating standards and the index of basically eliminating KBD proposed by the 12th Five-Year Plan.

Objective In order to accumulate data for the final evaluation and acceptance of the Twelfth Five-Year Plan of National Endemic Disease Prevention and Control,we investigated the endemic scopes and basic situation of Keshan disease (KD) in Harbin.Methods According to the Criteria for Diagnosis of Keshan Disease (WS/T 210-2011),we conducted clinical survey of KD prevalence in 10 KD endemic counties in Harbin in 2013.2011-2015,we detected and searched for cases in 10 ward counties of Keshan disease.According to the Delimitation and Classification of Keshan Disease Areas and the Evaluation of Endemic Disease Control and Elimination,we determined the regional type,the situation of control and elimination of the disease,at the same time all KD patients were positioned by global positioning system (GPS).Results There were 2 KD endemic counties,12 endemic townships,and 37 KD patients in Harbin.All 12 townships were the light endemic regions of KD.Other townships were conformed to be the history regions with KD.The management index scores of KD reached the elimination requirements which were more than 85 points in every region county (city).Ten region counties got to technical standards for KD elimination.Harbin basically reached the index of KD elimination.The GPS satellite positioning of KD patients were concentrated in the 45.1-45.5 degrees north latitude,and 127.7-128.3 degrees east longitude.Conclusion The regions of KD endemic areas have narrowed down and the prevalence rate has declined significantly in Harbin,which have reached the basic elimination standards of KD by the 12th Five-Year Plan.

Objective:To explore the application of high-throughput sequencing (HTS) technology in pathogens detection for spinal infection.Methods:From January 2019 to May 2020, a total of 41 patients including 31 males and 10 females with an average age of 59.7±11.9 years (29-75 years) were suspected of spinal infections. There were 37 patients with local pain, 15 with fever (≥38 ℃) and 18 with neurological dysfunction. The infected sites were as follows, 4 cases of cervical spine, 8 cases of thoracic spine and 29 cases of lumbar spine. There were 36 patients met the surgical indications and underwent open debridement, bone grafting, fusion and internal fixation, while the other 5 patients underwent conservative treatment (three received drug therapy and two were transferred to the internal department for chemotherapy). Lesions obtained from open surgery patients were underwent pathology and HTS examination. In 5 cases with conservative treatment, two of them underwent CT guided percutaneous puncture for samples, while one case underwent ultrasound guided percutaneous puncture for pus, one case for venous blood, and one case received lumbar puncture for cerebrospinal fluid. The samples were sent for pathological and HTS examination, while liquid specimens were sent for bacterial culture and HTS. The sensitivity and specificity of HTS results were determined according to pathological examination which was regarded as the "gold standard". Based on HTS results combined with the clinical manifestations, imaging examination and pathological results of the patients, targeted antibiotics or anti-tuberculosis drugs were selected for postoperative drug therapy. Patients with bacterial infection received anti-infection treatment for 3 months after operation. For tuberculosis patients, "tetrad" (isoniazid+rifampicin+pyrazinamide+ethambutanol) anti-tuberculosis treatments were underwent for one year. Inflammation indicators from the blood samples were observed before and after treatment, including white blood cell count (WBC), C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). These indicators were used to monitor disease progression and the curative effects. All patients were followed up for at least 3 months after surgery.Results:A total of 41 patients with suspected spinal infection were included in this study. The HTS pathogen detection results were obtained within 48 h. For the initial 5 patients, first-generation sequencing verification was conducted with coincidence rate 100%. Further, no further verification was conducted in the rest patients. Among the 41 cases, a total of 26 cases had positive results with a positive rate of 63.4%(26/41). Among them, thirteen cases were with mycobacterium tuberculosis (31.7%) and 6 cases with staphylococcus (14.6%). Fungi and Brucellosis were diagnosed in 2 cases respectively, accounting for 4.9% respectively. The test were negative in 15 patients (36.6%), including 2 patients with tumor or tumor-like lesions (1 hematologic tumor and 1 eosinophilic granuloma). A total of 38 patients underwent pathological examination, which confirmed 7 cases of suppurative infection, 12 cases of tuberculosis, 2 cases of tumor or tumor-like lesions and the remaining 17 cases of inflammatory lesions. The sensitivity and specificity of HTS were 80%(16/20) and 55.6% (10/18) with positive predictive value (PPV) 66.7% (16/24) and negative predictive value (NPV) 71.4% (10/14). All patients were followed up for 3 months. The inflammation indicators of blood at 3 months were all lower than that at admission. WBC decreased from (7.50±3.26)×10 9/L at admission to (6.22±2.53)×10 9/L at 3 months after treatment without statistically significant difference ( t=1.082, P=0.290). The CRP decreased from (32.2±34.1) mg/L to (4.5±10.5) mg/L, and ESR from (44.2±26.5) mm/1 h to (18.6±12.1) mm/1 h with statistically significant difference ( t=8.963, P<0.001; t=5.421, P<0.001). Conclusion:High-throughput sequencing technology can be used in detection of spinal infection pathogens, due to its relatively high positive rate, satisfied sensitivity and good diagnostic value.

Aim To observe the protective effects of probiotics on alcoholic liver injury in rats .Methods Male Wistar rats were randomly divided into the follow-ing three groups: control group , normal diet with nor-mal (5 ×108 CFU· kg -1· d -1) treatment group.Ex-cluding the rats in the normal control group , the other animals were initially received intragastric administra-tion with 56%( V/V) ethanol 5.5~11.0 mL· kg -1 · day -1 for 8 weeks.Then the rats’ faeces were collect-ed, and the liver and the small intestine were obtained for pathologic and ultrastructural observation .Serum ALT, AST and ALP was measured by method of bio-chemistry .Serum DAO and D-LA was measured by en-zyme linked immunosorbent assay .The expression of FOXO4 in small intestine was detected by immunohis-tochemistry .The intestinal flora genome DNA was ex-tracted from faeces and the sequence of 16 S rDNA was analyzed by high-throughput sequencing technologies . Results Hepatic steatosis was obviously improved in probiotics treatment groups compared with ethanol-trea-ted group , and the ultrastructural such as mitochondri-al and rough endoplasmic reticulum pathological chan-ges was significantly alleviated . The ultrastructural changes in intestinal were better in probiotics treatment group than in the ethanol-treated group .And ethanol-induced rats ’ serum ALT, AST, ALP, D-LA and DAO levels showed a significant reduction in the probi-otics treatment groups compared with the ethanol-trea-ted group ( P<0.05 ) .The FOXO4 expression was in-creased obviously in the probiotics treatment groups compared with the ethanol-treated group ( P <0.05 ) . And the intestinal flora diversity was impacted after feeding alcohol , and probiotics had a certain regulative action in helping the intestinal flora back to normal state; At phylum level , the Firmicutes quantity was lower and the Bacteroidetes quantity was higher in eth-anol-treated group than those in the control group ( P<0.05 ) , and the conditions were improved after supple-menting probiotics .At genus level , the percent of ge-nus abundance was similar to normal control group in the probiotics treatment groups compared with the etha-nol-treated group .Conclusion Probiotics can relieve liver injury induced by alcohol in rats , and the mecha-nism may be related to the modulation of probiotics on the intestinal flora distribution and intestinal barrier .

Objective To explore the risk factors of postoperative liver dysfunction in cirrhotic patients with Child-Pugh A liver function after splenorenal shunt and esophagogastric devascularization.Methods 128 Child-Pugh A cirrhotics undergoing splenorenal shunt and portal-azygous disconnection in our hospital from February,2014 to February,2016 were evaluated for risk factors of postoperative liver dysfunction.Results 22 cases (17.2％) developed postoperative liver dysfunction.By single factor analysis,age,history of upper gastrointestinal hemorrhage,ascites,serum prealbumin,serum albumin,mechanical ventilation time after surgery,postoperative serum albumin were associated with postoperative liver dysfunction.Non-conditional Logistic regression showed that age ＞ 60,serum prealbumin ＜ 170 g/L and serum albumin ＜ 30 g/L was independent risk factors for postoperative liver dysfunction.Conclusion Patients' age ＞ 60,serum prealbumin ＜ 170 g/L and serum albumin ＜ 30 g/L was independent risk factors for postoperative liver dysfunction after splenorenal shunt and portal-azygous disconnection.

Objective: To investigate the long-term outcomes of posterior long segment instrumentation and fusion in adult degenerative scoliosis (ADS), and to explore the correlative factors of sagittal imbalance after long segment instrumentation and fusion, and to summarize the impacts of different lowest instrumented vertebra (LIV) on sagittal complication rates. Methods: ADS patients who underwent long segment instrumentation and fusion between January 2008 and January 2014 were retrospectively reviewed and divided into two groups depended on LIV (L ₅ group and S₁ group). The follow-up time was at least five years. Radiographic parameters, Visual Analogue Score (VAS), Oswestry Disability Indexscore(ODI), the 12-item Short Form Health Survey-Physical Component Summary (SF-12 PCS)and sagittal complications were analyzed. And the correlation between sagittal imbalance and sagittal complications were calculated. Patients were divided into two groups(sagittal imbalance group and sagittal balance group) to explore the correlative factors of sagittal imbalance. Results: All of 56 patients were included and evaluated in this study, and 35 cases stopped at L ₅, 21 cases at S₁. The mean follow-up time was 6.6±1.6 years. No significant difference were found in age, gender, follow-up time and surgery method (P>0.05) between two groups. No significant differences were found in Cobb angle(11.8°±7.3° vs 12.6°±6.4°), coronal balance distance(CBD) (0.7±0.6 cm vs 1.0±0.5 cm), sagittal vertebral axis(SVA)(3.0±3.1 cm vs 2.6±5.6 cm), thoracic kyphosis(TK) (21.4°±9.6° vs 22.5°±6.9°), lumbar lordosis(LL) (-32.7°±7.3° vs-34.8°±9.1°), pelvic tilt (PT) (23.4°±9.8° vs 23.3°±8.1°), VAS score(2.0±1.2 vs. 2.2±0.9), ODI score(18.5%±6.4% vs 19.3%±5.2%) and SF-12 PCSscore(45.5±5.2 vs 42.4±7.0) between two groups at final follow-up time(P>0.05). Radiographic parameters including Cobb angle, CBD, SVA and health related quality of life scores including VAS score, ODI score and SF-12 PCS scores were improved significantly at two weeks postoperatively, two-year follow-up time andfinal follow-up time in two groupscompared with those preoperatively (P<0.05). And no significant difference were found in radiographic parameters and health related quality of life scores between those at two years follow-up time and those at final follow-up time; Sagittal complication rates at final follow-up were higher than these at two years follow-up (82.1% vs 33.9%, P=0.000). Sagittal imbalance was related to internal fixation failure in L ₅ group (r=0.691, P=0.011), internal fixation failure in S₁ group (r=0.691, P=0.001) and pseudoarticulation formation (r=0.499, P=0.021). 10 patients in the sagittal imbalance group were with preoperative sagittal imbalance, 9 with preoperative coronal imbalance, 6 with internal fixation failure, 5 with proximal junctional kyphosis (PJK), 4 with adjacent segment degeneration during the follow-up periods, the incidence rates of which were higher than those in the sagittal balance group. LL in the sagittal imbalance group was lower than that in the sagittal balance group at two years follow-up time and final follow-up time. Compared with the sagittal imbalance group, the sagittal balance group achieved a higher SF-12 PCS (45.4±5.1 vs 41.6±7.4, P<0.05) and lower ODI scores (17.8%±6.1% vs 21.5±4.6%, P<0.05), and no significant difference was found in VAS scores between two groups (1.9±1.2 vs 2.4±0.8, P>0.05). The incidence of adjacent segment degeneration in L ₅ group was higher than that in S₁ group (P<0.05) at final follow-up time, and the incidence of pseudojoint in S₁ group was higher than that in L ₅ group (P=0.002) at final follow-up time. Conclusion Posterior long segment instrumentation and fusion for adult degenerative spinal deformity can obtain satisfying long-term clinical outcomes, and postoperative sagittal imbalance was related to preoperative sagittal/coronal imbalance, postoperativeinternal fixation failure, PJK, pseudoarticulation formation and adjacent segment degeneration. Distal fusion at L ₅ or S₁ may lead to high risk of adjacent segment degeneration or pseudoarticulation formation respectively.

At present, hepatic encephalopathy has a relatively high mortality and thus greatly affects patients’ quality of life. This article describes the changes of intestinal flora in patients with hepatic encephalopathy and analyzes the mechanism of action of intestinal flora in hepatic encephalopathy and related treatment methods. It is pointed out that the development of hepatic encephalopathy is closely associated with intestinal flora, and clinical treatment by regulating intestinal flora has achieved a marked effect in patients with hepatic encephalopathy. In the future, the research on intestinal flora in patients with hepatic encephalopathy can be deepened to provide better regimens for the treatment of hepatic encephalopathy.

Objective: To explore the relationship of sleep with C reactive protein (CRP) abnormality and hyperinsulinemia in adolescents, and to provide reference for early prevention of metabolic disorders. Methods: Based on the Chinese Metabolic Syndrome Cohort Study in Anhui Province, a total of 653 adolescents aged 12 to 19 were selected to examine the relationship between wakeup time, bedtime, sleep disordered breathing (SDB) and CRP abnormality as well as hyperinsulinemia. Results: Later wakeup time (OR=1.68, 95%CI=1.03-2.75) was positively correlated with a higher risk of fasting hyperinsulinemia. Late bedtime (OR=1.96, 95%CI=1.29-2.99) was associated with 2 h postprandial hyperinsulinemia. Among those with high CRP concentration, late wakeup time and late bedtime were positely associated with hyperinsulinemia than those with normal CRP concentration; and the correlation between SDB and hyperinsulinemia was observed. Conclusion Later wakeup and late bedtime may be risk factors for hyperinsulinemia in adolescents. High concentrations of CRP may further increase the risk of hyperinsulinemia, a condition associated with sleep problems.Teenagers should get up and go to bed as early as possible.