BACKGROUND: In the healing process of fracture, if the callus tissue around the location of fracture is cleared, the end of fracture is difficult to be integrated completely, and more or less there would be some bone de fect. We deduce that these callus tissues are the necessary product during the process of bone reparation, which ought to be useful. Concerning this,we observed histological content and the volume of transforming growth factor-β1 (TGF-β1) with histological analysis and hybridization in situ of the frozen section of these calluses to investigate the effects of callus graft in the healing process of fracture.OBJECTIVE: To understand the salvaging value of bony callus organization around fracture in delayed operation by means of determining histological substances and contents of TGF-β1.DESIGN: Hybridization in situ of the callus tissue and randomized-grouping and controlled experiment.SETTING: Department of Orthopaedics, the Second Hospital of Xi'an Jiaotong University, and Department of Orthopaedics, Central Hospital of Taizhou City PARTICHANTS: Totally 51 inpatients who received delayed fracture operations in the Department of Orthopaedics of the Second Hospital of Xi 'an Jiaotong University and Central Hospital of Taizhou City from July 1994 and October 2003 were recruited. In some cases of delayed fracture operation, callus was obtained with the consent of patients, Dig labeled kit was bought from Boehringer Mannheim Corp. CTT GCT GTA CTGTGT GTC CAA, TGF-β1 oligonucleotide probe sequence, was synthe sized by DNA synthesizer. Computer X-ray system was bought from Japan Fuji Corp. METHODS: 2,4,6,8 weeks after fracture, a small amounts of callus was obtained. Frozen section was made from undecalcified callus tissue, and noneisotope Dig labeled hybridization in situ, observe gene expression of TGF-β1 in callus. In the operation, after clearing the bone stump and fas tening the fracture, original bony callus was grafted to the bone defect and around the fracture. Simple callus graft, flank bone graft or both were adopted respectively on patients in delayed operation with follow-up obser vations were given for 1-4 years, with the average of 1 year and 9 months,at outpatient clinic for recheck.MAIN OUTCOME MEASURES: ① Histological substance and TGF-β1 content of bony callus at different phases of the fracture. ② Effect of different mode of bone graft on the time of fracture healing. RESULTS: Totally 51 cases were treated, and 7 cases were not followed up for death or moving to other areas. Data of the followed-up 44 patients were collected into the stage of result analysis. ① Gene expression of TGF-β1 in bone callus: In the second week after fracture, there were a small amount of fibrous bone callus and comparatively much cartilage bone callus,and TGF-β1 was hypso-expressed in cartilage cells inside the cartilage islet; about four weeks later, TGF-β1 was notably expressed in os teoblasts; about eight weeks later, a host of calluses grew, mainly carti laginea, and the numbers of formed fibroblast, callus, osteoblasts and tra becular bone were increasing. TGF-β1 expression in all kinds of cells dis appeared after six weeks but had a tendency to rise again in bone matrix in the eighth week. ② There were no significant difference of healing time in different modes of bone graft .CONCLUSION: TGF-β1 plays an important role in balancing bone healing, and necessary connections exist between tissue change in bone callus and change in expression of TGF-β1. As callus is a product of recovery and reconstitution of organism, we regard callus graft salvage as a feasible method in reducing pain and damage to patients who need graft.

Objective To observe the clinical efficacy of Zhishanghan Zhufeng Formula in treating knee osteoarthritis.Methods Totally 90 patients were randomly divided into treatment group and control group, 45 cases in each group. The treatment group was treated with Zhishanghan Zhufeng Formula for 30 days. The control group was treated with routine medicine (diclofenac sodium sustained release tablets and glucosamine sulfate tablets) for 30 days. The clinical efficacy and the changes of VAS, WOMAC score and knee temperature before and after treatment of two groups were observed.Results TCM clinical symptoms were significantly improved, and the effects in the treatment group were more evident compared with the control group (P<0.05). After treatment, VAS and WOMAC score of the two groups decreased obviously, while the knee temperature increased, and the treatment group was superior to the control group, with significant significance (P<0.05).Conclusion Zhishanghan Zhufeng Formula has effective clinical efficacy for knee osteoarthritis.

Objective To evaluate the efficacy of Jianpi-Xiaozheng recipe combined with chemotherapy in patients with late gastric cancer. Methods A total of 124 patients with late gastric cancer were randomly divided into a control group and a treatment group by random number table method, with 62 cases in each group. The patients in the control group received chemotherapy with S-1 and oxaliplatin (SOX), and those in the treatment group received Jianpi-Xiaozheng recipe combined with SOX chemotherapy. The treatment response was evaluated using the response evaluation criteria in solid tumors. The quality of life and physical status were evaluated with the European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire (QLQ-C30) and Karnofsky Performance Status (KPS), respectivly. The serum levels of tumor biomarkers, carbohydrate antigen 199 (CA199) and carbohydrate antigen 72-4 (CA72-4) were detected before and after treatment. Results The response rate (complete or partial response) in the treatment group was significantly higher than that in the control group (62.9%vs. 43.5%;χ2=4.665, P=0.031). The scores of QLQ-C30 (46.8 ± 6.3 vs. 42.2 ± 5.9;t=4.196, P=0.001) and KPS (79.1 ± 7.8 vs. 72.0 ± 7.5;t=5.167, P=0.000) in the treatment group were significantly higher than those in the control group. The serum levels of CA199 (61.7 ± 16.5 U/ml vs. 113.3 ± 21.4 U/ml;t=15.036, P=0.000) and CA72-4 (27.9 ± 9.6 U/ml vs. 34.3 ± 9.7 U/ml;t=3.693, P=0.001) in the treatment group were significantly lower than those in the control group. Conclusions Jianpi-Xiaozheng recipe combined with chemotherapy can increase response rate, decrease the serum levels of tumor biomarkers, and improve the quality of life in patients with late gastric cancer.

Objective The aim of this study is production of organ specific animal model for studying reproductive toxicity in mice.Methods F1 generation was gotten by mating the Ddx4 -cre transgenic male mice with the Rosa26mT/mG transgenic female mice.F1 offspring and its parents phenotype was screened by molecular biological, histopathological and in vivo imaging technology.Results At molecular level, specific DNA fragment was only found in testis of F1 offspring; At the organ level, the expression of green fluorescent protein could only be observed in testis of F1 offspring; Testicular frozen sections and sperm fluorescence observation showed that green fluorescent protein were mainly expressed in the germ cell lineage such as secondary spermatocyte and spermatocyte and spermatozoon.Conclusions The production of the mice with specific germ cell expressed green fluorescent protein by Cre/loxP recombination system were built successfully.

Background Multimorbidity imposes a heavy burden on individuals, families, and society. There are relatively few studies exploring patterns of multimorbidity among middle-aged adults in China. Objective To explore the current status of multimorbidity, associated risk factors, and multimorbidity patterns among adults aged 45-64 years in China, so as to provide a scientific basis to prevent and control multimorbidity in China. Methods A total of 5494 adults aged 45-64 years from the Chinese Health and Nutrition Survey (CHNS) in 2018 were selected. Of these, 2494 (45.39%) were men and 3000 (54.61%) were women. The nine diseases included were hypertension, diabetes, dyslipidaemia, obesity, mild cognitive impairment (MCI), myocardial infarction, stroke, asthma, and tumor. The prevalence of each disease or multimorbidity was expressed as N (%). Comparisons of multimorbidity prevalence between different groups were performed using the χ² test or Cochran-Armitage trend test. Association rule with the Apriori algorithm was used to explore the pattern of multimorbidity, with parameters set at a minimum conditional support of 3.00%, a minimum rule confidence of 50.00%, and a lift of >1.20. Logistic regression was used to evaluate the associations between selected risk factors and multimorbidity. Results In 2018, 37.44% of participants reported multimorbidity in 15 provinces of China. The prevalence of diseases in descending order was dyslipidaemia (39.99%), hypertension (39.48%), obesity (16.42%), MCI (14.47%), diabetes (14.16%), tumor (1.09%), stroke (1.04%), myocardial infarction (0.71%), and asthma (0.64%). A total of seven multimorbidity patterns were identified in this group. Obesity paired with hypertension, and diabetes paired with dyslipidemia were the two major patterns of multimorbidity in the general population and age or sex subgroups. The multimorbidity patterns of different populations were concentrated in the combination of obesity, hypertension, diabetes, and dyslipidemia. The risk of multimorbidity was lower in females than in males (OR=0.85, 95%CI: 0.75, 0.97). The multimorbidity risk was 1.56 times higher in the 55-64 years group than in the 45-54 years group (OR=1.56, 95%CI: 1.40, 1.75). Drinking in the past year increased the risk of multimorbidity by 25% (OR=1.25, 95%CI: 1.08, 1.45) compared to no alcohol comsumption in the past year. High and medium levels of physical activity were associated with a decreased OR (high: OR=0.74, 95%CI: 0.65, 0.85; medium: OR=0.81, 95%CI: 0.70, 0.93) with low level of physical activity as reference. Conclusion In 2018, there was a high prevalence rate of multimorbidity among middle-aged adults in China. The main multimorbidity patterns were obesity-hypertension and diabetes-dyslipidemia. Surveillance and interventions should be strengthened particularly for men, individuals with alcohol consumption or insufficient physical activity, and those with major multimorbidity patterns.

BACKGROUND: It has been reported that human adipose-derived mesenchymal stem cells can be induced to differentiate into hepatocyte-like cells with the function of albumin synthesis and urea secretion in vitro.OBJECTIVE: To investigate the potential of adipose-derived mesenchymal stem cells differentiating into hepatocyte-like cells in vitro.METHODS: Adipose-derived mesenchymal stem cells were isolated from the subcutaneous fat of hepatitis B virus infection patients by collagenase digestion and adherent method. Adipose-derived mesenchymal stem cells were induced by three-phase induction method and observed morphologically. The expression levels of alpha-fetoprotein, albumin and cytokeratin 18 were detected by immunohistochemical staining and glycogen synthesis function was detected by glycogen staining method.RESULTS AND CONCLUSION: Most of adipose-derived mesenchymal stem cells induced by three-phase induction method were differentiated into hepatocyte-like cells with polygonal morphology. Immunohistochemistry staining results showed that hepatocyte-like cells expressed alpha-fetoprotein, albumin and cytokeratin 18, and the expression levels of albumin and cytokeratin 18 increased with the culture time. The induced cells had the function of glycogen synthesis and were positive for periodic acid Schiff staining. These results showed that the subcutaneous adipose-derived mesenchymal stem cells could be induced into functional hepatocyte-like cells in hepatitis B virus infection patients.

Objective To analyze the cognitive changes and influencing factors in patients with lacunar cerebral infarction after carotid artery intervention therapy. Methods Sixty lacunar cerebral infarction combined with carotid stenosis patients treated with artery intervention therapy (intervention therapy group) and 68 lacunar cerebral infarction without carotid stenosis patients treated with drug therapy (drug therapy group) were selected. The neuropsychological test was completed at entry and 1, 6, 12 months after entry, and the results were compared with 60 healthy controls (control group). The cognitive changes were observed. The neuropsychological test included mini mental state examination (MMSE), Montreal cognitive assessment scale (MoCA) and cognitive field test. Results There were statistical differences in other scores except the Stroop test C section and Wechsler adult intelligence scale (WAIS-RC) picture arrangement subtest at entry in intervention therapy group and drug therapy group compared with control group (P<0.05). There were no statistical differences in the all scores at entry between drug therapy group and intervention therapy group (P>0.05). In intervention therapy group, the MMSE scores, MoCA total score, Rey-Osterrieth complex figure test (ROCFT), auditory verb learning test (AVLT), and the WAIS-RC picture arrangement subtest, verbal fluency test, WAIS-RC digit span backwards subtest of performing function 12 months after entry were significantly better than those at entry, and there were statistical differences (P<0.05). MMSE score, MoCA total score, long-time delayed recall of ROCFT, the immediate recall, long-time delayed recall and short delayed recall of AVLT, semantic category fluency test of performing function and digit span backwards subtest of WAIS-RC 6 months after entry were significantly better than those at entry:(27.8 ± 2.2) scores vs. (26.4 ± 1.9) scores, (20.7 ± 2.3) scores vs. (19.3 ± 2.0) scores, (12.4 ± 3.2) scores vs. (10.8 ± 2.6) scores, (54.3 ± 10.6) scores vs. (49.9 ± 10.9) scores, (12.4 ± 2.0) scores vs. (11.2 ± 2.8) scores, (12.9 ± 2.0) scores vs. (10.6 ± 2.6) scores, (17.5 ± 4.0) scores vs. (15.4 ± 3.4) scores and (4.0 ± 0.9) scores vs. (3.5 ± 0.9) scores, and there were statistical differences (P<0.05). In drug therapy group, there were no statistical differences in the all scores 1 and 6 months after entry, compared with that at entry (P>0.05);the MMSE score, MoCA total score, ROCFT, the immediate recall, long-time delayed recall and short delayed recall of AVLT, WAIS-RC picture arrangement subtest, verbal fluency test, WAIS-RC digit span backwards subtest of performing function and digit span backwards subtest of WAIS-RC 12 months after entry were significantly better than those at entry, and there were statistical differences (P<0.05). There were no statistical differences in all scores 12 months after entry between intervention therapy group and drug therapy group (P>0.05). In patients intervention therapy group, Logistic regression analysis showed that the MoCA score was related with age, hypertension and low education level (P<0.01 or<0.05), but was not related with smoking, diabetes and interventional treatment (P>0.05). Conclusions Cognitive impairment in patients with lacunar cerebral infarction and carotid stenosis is severe and extensive, but most cognition disorders can improve to normal level 12 months after artery intervention therapy.

BACKGROUNDTo explore the characteristics and clinical significance of diffusing capacity in the patients with lung cancer.

METHODSThe pulmonary diffusing capacity for carbon monoxide (DLCO) was measured with the rebreathing method in 138 patients with primary lung cancer, and 86 were performed pulmonary resection.

RESULTSThere was no significant difference in DLCOrb and DLCOrbc between the groups with different histological type and degree of ventilation impairment and general type (P > 0.05). DLCOrb/VA mildly decreased in the patients whose lung ventilation function was normal. DLCOc/VA decreased in the patients with restrictive ventilation dysfunction (P < 0.01), and DLCOrb/VA decreased in the patients with obstructive and mixed ventilation dysfuncion (P < 0.05). DLCOc/VA decreased in the patients with light and obvious lung dysfunction (P < 0.05). DLCOc/VA in the patients with central lung cancer was lower than that in the peripheral ones (P < 0.01). DLCOc/VA and D LCOrb in the group with postoperative respiratory failure were lower than that in the group without respiratory failure (P < 0.05). When DLCOc/VA of less than 80% and DLCOrb/VA of less than 70% were used to predict the postoperative respiratory failure, the correct ratios of dignosis and the diagnostic indexes were high.

CONCLUSIONSThe diffusing capacity decreases in the patients with lung cancer, and the main manifestation is the abnormal DLCOrb/VA and DLCOc/VA. The general type of lung cancer and the degree of pulmonary dysfunction may influence the diffusing function of the patients. DLCOc/VA of less than 80% and DLCOrb/VA of less than 70% should be selected for predicting postoperative respiratory failure.

Objective:To highly express HIV-1p17 in E.coli and purify the protein.Methods:①Recombinant plasmid was constructed by inserting HIV-1p17 gene amplified by PCR into plasmid vector,pET28c;②The recombinant plasmid was expressed in BL21,BL21(DE3),BL21(DE3) plysS and HMS174(DE3) of E.coli separately;③The target protein were purified with Ni-NTA resin;④The purified protein was detected by western blot and ELISA.Results:The expression of the P17 protein in BL21(DE3) represented up to 32% of total protein in E.coli,which was the most amounts compared with other kinds of E.coli.The purity of the purified protein reached 95%.The purified protein was recognized by HIV-1P17McAb as well as by HIV-1 positive serum.Conclusion:The recombinant plasmid is highly expressed in BL21(DE3) of E.coli that can be proceeded to the immunocompetence and the bioactivity research.The method of Ni-NTA resin is simple with low protein losing and high purity.And the purified p17 can be employed in early detection of HIV-1 infection and prediction of the clinical progression.

Objective Respectively applying the treatment of biventricular pacing and right ventricular septal pacing in atrioventricular block,to compare the heart function influence of two kinds of pacing mode on pacemaker dependent patients, to provide evidence for the physiological pacing mode selection?Methods Enrolled 20 patients from January 2012 to March 2013 who should be placed in pacemakers, their primary disease was the second degree,high or third degree atrioventricular block,giving them three chamber pacemaker ( right atrial + biventricular ) each?Randomly divided into right ventricular septum pacing group ( group A, n=10) and biventricular pacing group( group B,n=10)?Twelve months later,each group crossed into the each other group and continued following?up for 12 months?After 24 months to obtain all the data to do the statistical analysis,including patients'6 min walking distance(6MWD),the Minnesota Heart Failure Quality of life score (MLHFQ),plasma N?terminal pro brain natriuretic peptide precursor(NT?proBNP),left ventricular ejection ejection fraction(LVEF),left ventricular diastolic end diastolic diameter(LVEDD),left ventricular contraction end diastolic diameter(LVESD),left ventricular twelve segmental 14W time standard deviation(Ts?12SD),left ventricular twelve segmental 14W time maximum delay(Ts?dif),the paced QRS qrsd?Results Compared with group B,the 6MWD and LVEF of 12,24 months after treatment of group A were significantly increased( ( 242?58 ±37?56) m vs?(347?42±36?59) m vs?(340?67±24?99) m;(39?97±5?84)% vs?(57?92±10?01)% vs?(60?50±10?06)%;P<0?05),QRSd and NT?proBN were significantly decreased((139?25±10?43) ms vs?(114?25±10?07) ms vs?(110?83±11?08) ms) ms;( 2 857?84±236?48) ng/L vs?( 2 144?26±301?43) ng/L vs? (2 025?91±307?42) ng/L;P<0?05)?Compared with before treatment,at 12 and 24 months after treatment,6MWD in group B was significantly increased(228?17+38?06) m,(329?33+46?28) m,(350?67+35?43) m, LVEF was significantly increased ( ( 40?25+11?24 )% vs? ( 59?50+9?14 )% vs? ( 60?17+10?29)%),QRSd significantly narrowed((142?42+10?66) ms vs?(118?58+994) ms vs?(116?25+10?59) ms) and NT proBNP levels significantly reduced((2 848?25+318?65) ng/L vs?(2 144?26+301?43) ng/L vs?( 2 025?91+30?742) ng/L) were,the difference had statistical significance ( P<0?05)?There was no significant difference on the different time between the groups ( P=0?05)?Comparisons between A and B group at the same treatment time,these indexes of detections were no statistical significance(P>0?05)?Conclusion Compared with the right septal pacing,biventricular pacing is of no significant advantages on the effect of cardiac function for patients with pacemaker dependent.