Objective To investigate the relation between serum leptin/tumor necrosis factor-α (TNF-α)and malnutrition in patients with chronic obstructive pulmonary disease (COPD) and stable chronic cor pulmonale (CCP) at high altitude. Methods Totally 162 COPD and CCP patients and 40 normal controls (group C) were studied. COPD and CCP patients were divided into malnutrition group (group A, n = 104) and normal nutrition group (group B, n =58) according to the nutritional parameters. Levels of serum leptin and TNF-α were measured by enzyme-linked immunosorbent assay (ELISA). Results Body mass index (BMI), percentage of normal body weight (NW%), triceps skinfold thickness (TSF), mid-upper arm circumference (MAC), serum albumin (ALB) ingroupA[(17.4±1.8) kg/m2, (82.3±4.3)%, (7.0±2.6) mm, (17.8±2.8) cm, (30.3±3.9)g/L, respectively] were significantly lower than those in group B and group C [(21.8 ± 2.0) kg/m2,(98.6±5.5)%, (9.3±2.6) mm, (21.5±2.9) cm, (36.2±3.8) g/L, and (23.1±2.3) kg/m2,(102.2±5.2)%, (9.7±3.8) mm, (22.1±2.8) cm, (36.8±3.9) g/L, respectively; all P＜0. 01].The levels of serum leptin and TNF-α in group A [(9.5 ±1. 8) ng/ml and (17.3 ±2. 2) ng/ml, respectively]were significantly higher than those in group A and group C [(7.3 ± 2. 0) ng/ml, (13.5 ± 2. 3) ng/ml; and (6. 7 ±2. 3) ng/ml, (12. 8 ±2. 1) ng/ml, respectively; all P ＜0.01). However, they were not significantly different between group A and group B (all P ＞ 0. 05). The level of leptin was negatively correlated with BMI (r=-0.745, P=0. 0005), NW% (r= -0.887, P=0. 0005), TSF (r= -0.725, P=0. 0005), MAC (r= -0. 761, P=0. 0005), serum albumin (r= -0. 558, P=0. 0005) in group A, and was positively correlated with TNF-α (r = 0. 527, P = 0. 0005). Conclusion Serum leptin and TNF-α correlate with malnutrition in patients with COPD and CCP at high altitude.

Objective To explore the mechanism of preventive effect of Chinese medicine with Replenishing kidney and invigorating Qi on osteoarthritis. Methods 72 male Long-eared white rabbits aged 4 months were randomly divided into six groups, A group (blank control group), B group (model group), C group (traditional Chinese medicine high-dose prevention group), D group (Chinese herbal medicine with medium-dose prevention group), E group (Chinese herbal medicine with low-dose prevention group), F group (glucose-amino acid hydrochloride capsules prevention group).All the animals apart from A group were established osteoarthritis model by immobilizing knee joint with plaster cast for 6 weeks. In the same time of immobilization, traditional Chinese medicine and glucose-amino acid hydrochloride capsules were given to C, D, E and F group for 4 weeks. Physiological saline was given to B group. After 6 weeks of modeling, synovial fluid was extracted and the changes of TNF-α in it were analyzed by ELISA. Results Articular cartilage degeneration of B group was most obviously compared with C, D, E and F group. There was a significant difference of TNF-α level in comparison of A and B group (P＜0.05), B and C, D, E and F group (P＜0.05), and among C, D and E group. Conclusion Chinese medicine with replenishing kidney and invigorating Qi can prevent osteoarthritis by reducing TNF-α level in synovial fluid, enhancing cartilage cell metabolism, and slowing down cartilage degradation.

Objective To investigate the expression of miR-424* in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of non-small cell lung cancer(NSCLC) patients,and to explore its potential role in the diagnosis and prognosis of lung cancer.Methods A549 cells were irradiated with 2 and 4 Gy X-rays,and some of irradiated cells were injected into nude mice through tail vein.Real time quantitative PCR (RT-qPCR) assay was employed to detect the expression of miR-424 * in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of lung cancer patients.Results Compared with the control group,the expression of miR-424* was up-regulated significantly in X-ray irradiated A549 cells at 1,2,12,24 and 48 hpost irradiation,respectively (2 Gy:t =-45.886--6.709,P ＜0.05;4 Gy:t =-29.087--7.833,P ＜ 0.05).Furthermore,the expression of miR-424 * was up-regulated in the lung and serum of nude mice with injection of 0,2 and 4 Gy X-ray irradiated A549 cells,compared with control group (fold change was 9.72,8.58 and 4.7 with 2 Gy irradiation and 11.93,9.22 and 8.99 with 4 Gy irradiation,t=-13.243,-12.409,-9.833 in lung andt=-6.436,-3.052,-3.609 in serum,respectively,P ＜ 0.05).Out of 11 tissue samples of NSCLC patients,6 were detected with up-regulated miR-424* expression,and no significant discrepancy of miR-424* expression was detected in two type of NSCLC tissue samples.On the contrary,43 serum samples were detected with up-regulated miR-424* expression out of 84 serum samples (51.20％) of NSCLC patients (fold change range 1.97 to 17.71),and significant discrepancy of miR-424* expression was shown in two subtypes of NSCLC serum samples [adenocarcinoma:39.10％ (18/46) and squamous carcinoma:65.8％ (25/38)],as well as in serum samples of NSCLC patients with radiotherapy [41.5％ (22/53)] and without radiotherapy [67.7％ (21/31)] (t=5.919,5.387,P ＜0.05,respectively).Conclusions 2 and 4 Gy X-ray irradiation could up-regulate the expression of miR-424* in A549 cells,which might be correlated with the enhanced metastasis of A549 cells induced by X-ray in vivo and in vitro.Furthermore,the expression of miR-424* was up-regulated in over 50％ of the tissue and serum samples of NSCLC patients,which might be correlated with the diagnosis of NSCLC subtype and prognosis of radiotherapy.

Liver cancer is a malignant tumor with the characteristics of high morbidity and mortality.However,the routine detection methods for the liver cancer such as imaging tests and pathological detection are with low specificity and sensitivity.And it's also hard to detect the condition of patients dynamically.Cell-free DNA (cfDNA) is the DNA fragment that is away from the extracellular and exists in the blood,which contains genetic information from the organism.The related disease information could be reflected on the cfDNA in patients with liver cancer,indicating that cfDNA has great potential in the screening,diagnosis,treatment and prognosis of liver cancer.The clinical applications of cfDNA can make up for the deficiency of existing detection methods and achieve noninvasive and dynamic monitoring of liver cancer.This article will present a review on the progress of clinical applications of cfDNA in liver cancer.

Objective To establish genotyping methods for vitamin K epoxide reductase complex subunit 1 (VKORC1)and cytochrome P450 2C9(CYP2C9)based on pyrosequencing technique to detection of warfarin metabolizing enzyme related gene polymorphisms.Methods A total of 50 peripheral blood samples from healthy adults were collected and the whole blood genomic DNA was extracted.A set of biotin-labeled amplifi-cation primers and sequencing primers were designed respectively for three SNP sites:VKORC1 -1639 G>A,CYP2C9 430C> T and CYP2C9 1075A>C.After PCR amplification of the samples,pyrophosphoric acid se-quencing was conducted.And then the signal peaks form were combined to analyze and determine each sample genotype.Genotyping results were verified by Sanger sequencing,and the consistency of the two sequencing methods was compared.Results Genotypes of the three SNPs can be clearly determined according to the ba-ses and height of the signal peaks.Among the 50 samples,there were 41 AA and nine AG for VKORC1 -1639G>A,accounting for 82% and 12% respectively,and there were 45 *1/*1,five *1/*3 for CYP2C9, accounting for 90% and 10% respectively,no CYP2C9*2 allele detected.Genotype results detected by pyrose-quencing and Sanger sequencing were consistent with each other.Conclusion In SNP genotyping,Pyrose-quencing has the advantages of convenience,time-saving,cheap with accurate and reliable results,which can quickly determine the genotypes of CYP2C9 and VKORC1.

Objective: To improve physical activity by assessing pedometer-based physical activity during specific intervals over a week, among junior high school students,and to provide a reference for improving the level of students’ physical activity. Methods: Students (n=675) of grades 6-8 from six public junior high schools in Shanghai were recruited and instructed to wear a pedometer for a week, step counts and contents were recorded over a specific period. This period mainly included steps taken on the way to and from school, at school, at home in the evening on weekdays, and in the morning, afternoon, and evening on weekends. Results: The daily step counts recorded were 8 332 steps, with those on weekdays showing significantly higher values than step counts on weekends (9 065 steps vs. 6 392 steps)(t=22.9, P<0.01). Proportionately, the physical activity level at school contributed more to daily step counts (61.3%), followed by those on the commute to and from school(25.5%). Boys were more active than girls. For all intervals on weekdays, the step counts of students in rural districts, with overweight or obese, in grades 6 and 7, were higher than those in urban districts, with normal status, in grade 8 respectively (P<0.05). Low-activity students with physical education were more active than one without physical education(t boy=1.99,t girl=2.45,P<0.05). Conclusion These findings facilitate the implementation of effective, feasible interventions to enhance physical activity over a series of intervals during the day.

Objectives:To compare the effects of 3D-printed personalized guiding template and robot-assisted pedicle screw placements in orthopedic surgery for adult degenerative scoliosis(ADS).Methods:Retrospective analysis was conducted on 18 ADS patients hospitalized and treated with corrective surgery in the department between January 2020 and December 2022.There were 3 males and 15 females,aged 46-73(63.2±8.2)years old.A total of 236 pedicle screws were placed,and the patients were divided into two groups according to the auxiliary screw placement methods:the 3D-printed personalized guiding template group(3D-printed group,1 1 cases,142 pedicle screws were placed)and robot-assisted screw placement group(Robot group,7 cases,94 pedicle screws were placed).The patients were followed up for 6 months and more.The vertex rotation angle and scoliosis Cobb angle were measured and compared between groups before operation.All pedicle screws were classified by Gertzbein-Robbins classification standard at 1 week after operation,and the accuracy and satisfaction of screw placement in the two groups were compared;And the operative time and incidence of complications were also analyzed and compared between the two groups.Re-sults:The 3D-printed group was not statistically different from the robot group in terms of age(63.6±9.0 years vs.62.6±7.3 years),gender ratio(male/female:2/9 vs.1/6),number of screws placed(12.91±3.83 vs.13.43± 3.60),Cobb angle(40.36°±1 1.82° vs.38.14°±12.84°),and vertex rotation angle(30.27°±7.25° vs.29.86°±9.65°),respectively(P＞0.05).The robot group was longer in operative time than that of 3D-printed group(354.29± 53.73min vs.282.27±73.87min,P＜0.05).Of the total 142 pedicle screws placed in the 3D-printed group,128 screws were of class A,10 were of class B,and 4 were of class C;Of the total 94 pedicle screws placed in the robot group,86 screws were of class A,5 were of class B,and 3 were of class C;No class D or E screws in the two groups.The accuracy rate of screw placement(90.14％)and satisfaction rate(97.18％)of the 3D-printed group were not statistically different from the accuracy rate(91.49％)and satisfaction rate(96.81％)of the robot group,respectively(P＞0.05).The incidence of complications in the 3D-printed group(36.36％)was not significantly different from that in the robot group(57.14％,P＞0.05).Conclusions:Both auxiliary screw placement methods can assist spinal surgeons to accurately place screws in ADS patients,but 3D-printed personalized guiding template needs less time in screw placement.

Objective To investigate the role of BMP-2/Smad signaling pathway in the osteogenic differentiation of human aortic smooth muscle cells (HASMCs) caused by hyperphosphatemia -induced calcium phosphate (CaP) crystals.Methods High-phosphate medium was incubated at 37℃ for 3 days.CaP crystals and supernatant were isolated by ultracentrifugation.Scanning electron microscope and energy dispersive X-ray spectroscopy were performed for analysis of physicochemical characteristics of CaP crystals.HASMCs were cultured in vitro,and divided into high-phosphate,control,crystals and supernatant groups.Calcification was visualized by Alizarin red staining.Calcium loads in cells were quantified by o-cresolphthalein complexone method.Protein expression of bone morphogenetic protein-2 (BMP-2),Runt-related transcription factor 2 (RUNX2),osteopontin (OPN),phospho-Smad1/5/9 (p-Smad1/5/9) were quantified by Western blotting.After knockdowns of BMP-2 and Smad1 with small hairpin RNA (shRNA) interfering respectively in HASMCs,protein expressions were measured by Western blotting.Results High-phosphate medium induced the formation of CaP crystals.Compared with the cells in control group,CaP crystals significantly induced HASMCs calcification,increased calcium loads and up-regulated the levels of BMP-2,RUNX2 and OPN proteins (all P ＜ 0.05).After the addition of CaP crystals into HASMCs,the level of p-Smad 1/5/9 protein peaked at 30 min (P ＜ 0.05).After BMP-2 was knocked down in HASMCs,the expression of p-Smad1 caused by CaP crystals was blocked completely,and the expressions of RUNX2 and OPN caused by CaP crystals were reduced significantly (all P ＜ 0.05).After Smad1 was knocked down in HASMCs,the expressions of RUNX2 and OPN caused by CaP crystals were decreased significantly (all P ＜ 0.05).Conclusions Hyperphosphatemia-induced CaP crystals promoted osteogenic differentiation of HASMCs through the BMP-2/Smad signaling pathway.

Objective:To investigate the predictive value of peritoneal protein clearance (Pcl) for cardiovascular events and cardiovascular mortality in peritoneal dialysis (PD) patients.Methods:Eligible PD patients were prospectively enrolled from January 2014 to April 2015 in the PD Center of Renji Hospital, School of Medicine, Shanghai Jiao Tong University. All patients were followed up until death, withdrawing from PD, transferring to other centers, or the end of study period (October 1, 2018). The patients were divided into high Pcl group and low Pcl group by the median Pcl, and the differences of related indicators between the two groups were compared. A multiple linear regression model was used to analyze the influencing factors of Pcl. The Kaplan-Meier method and Log-rank test were used to compare the cumulative survival rates of patients between the two groups. A multivariate Cox regression model was used to estimate the risk of cardiovascular events and cardiovascular mortality in relation to Pcl in PD patients.Results:A total of 271 patients were enrolled, with 135 males (49.8%), age of (56.92±0.84) years old and a median PD duration of 38.77(19.00, 63.10) months. There were 70 patients (25.8%) comorbiding with diabetes and 81 patients (29.9%) with cardiovascular diseases (CVD). The median Pcl of this cohort was 67.93(52.31, 88.36) ml/d. Compared with the low Pcl group (Pcl<67.93 ml/d), the high Pcl group (Pcl≥67.93 ml/d) had older age, and greater proportion of CVD, body mass index (BMI), pulse pressure, brain natriuretic peptide, mass transfer area coefficient of creatinine (MTACcr), and lower serum albumin (all P<0.05). There was no significant difference in gender, dialysis duration, proportion of diabetes, proportion of angiotensin converting enzyme inhibitor and angiotensin receptor blocker, proportion of continuous ambulatory PD, high sensitivity C reactive protein, fluid removal including 24 h urine volume and 24 h ultrafiltration, and residual renal function between the two groups (all P>0.05). Multiple linear regression analysis showed that serum albumin ( β=-0.388, P<0.001), BMI ( β=0.189, P<0.001), and MTACcr ( β=0.247, P<0.001) were independently related to lg(Pcl). During the study period, 55 patients experienced one or more cardiovascular events and 39 patients had cardiovascular mortality. According to Kaplan-Meier analysis, cardiovascular mortality in the high Pcl group was higher than that of low Pcl group (Log-rank χ2=6.902, P=0.009). Multivariate Cox regression analysis showed that, high lg(Pcl) was an independent influencing factor of cardiovascular events in PD patients ( HR=7.654, 95% CI 1.676-34.945, P=0.009). Conclusions:Serum albumin, BMI and MTACcr are independently associated with Pcl, and Pcl is an independent predictor of cardiovascular events in PD patients.

Objective: To evaluate the accuracy of endoscopic titanium clip localization combined with CT three-dimensional reconstruction for the control of incision margin in early gastric cancer under laparoscopy. Methods: A prospective analysis was made for gastric cancer whose lesions were located in the middle of the stomach and T stage was 1 to 2 from October 2017 to January 2019 at Department of Gastrointestinal and Pancreatic Surgery, Zhejiang Provincial People′s Hospital. Totally 25 patients were eventually enrolled in the study. There were 17 males and 8 females aging of (63.6±7.2) years (range: 48 to 77 years). All cases were treated with titanium clip localization under endoscope combined with CT three-dimensional(3D) reconstruction to construct a virtual panorama of gastric cavity and lesions, and to design surgical margins. Laparoscopic surgical resection was performed according to the surgical margins designed before operation. The distance from the gastric angle to the origin of the minor curvature of the incisional margin, the distance from the gastric angle to the the center of lesion and the distance of the upper incision margin were measured under three-dimensional CT reconstruction and under actual specimen. Paired t test was used to compare the three distances measured by two methods. Results: The measured distances from the gastric angle to the center of the lesion and the proximal incisional margin under 3D reconstruction CT were according to the measured values of actual specimens ((2.67±1.38) cm vs. (2.83±1.56) cm, t=1.51, P=0.14; (5.23±0.60) cm vs. 5 cm, t=1.93, P=0.07); the measured distances from the gastric angle to the origin of the minor curvature of the incisional margin under CT 3D reconstruction were different with the measured values of solid specimens ((5.94±0.94) cm vs. (6.37±0.90) cm, t=3.52, P=0.00). Conclusion The method of titanium clip localization combined with CT 3D reconstruction can provide a feasible laparoscopic localization method and incision edge solution for T1 to T2 gastric central cancer.