Objective:To investigate the effect of adenosine (Ado) on the unit discharging electricities in habenula nucleus and on the c-fos expression in lateral habenular complex,and the influence of adenosine on the neuron activities and related gene expression involved in affecting sleep in habenula nucleus and the possible mechanisms.Methods:Intraperitoneal injection,brain flakes pouring of rats,immunohistochemistry and other methods were useel.Results:Ado pouring into flakes of brain depressed the unit discharging electricities of neurons in medial habenular complex(MHb),but obviously increased that of lateral habenular complex(LHb).0.5 h after the six rats being injected intraperitoneally with Ado,the c-fos protein expression in lateral habenular nucleus was significantly increased compared to the group with saline injection.Conclusion:Ado may restrain the unit discharging electricities of neurons in medial habenular complex but excite those in lateral habenular complex.At the meantime,Ado may increase the c-fos expression in LHb.This provides the experimental evidence that Ado may improve the sleep quality.

Objective To investigate the effect of tanshinone ⅡA on growth and apoptosis in human hepatoma cell line BEL 7402 in vitro . Methods The human hepatoma cell line BEL 7402 was treated with tanshinone ⅡA at various concentrations for 72 h. Cytotoxicity was evaluated by MTT assay, apoptosis related alterations in morphology ascertained by cytochemical staining(Hoechst 33258) and transmission electron microscopy(TEM). Apoptotic rate was quantified by flow cytometry (FCM). Results Tanshinone ⅡA inhibited the growth of hepatoma cells in a dose dependent manner, with IC 50 values of 6.28 ?g/ml. After treatment with 1～10 ?g/ml tanshinone ⅡA for 72 h, BEL 7402 cell apoptosis with nuclear chromatin concentration and fragmentation as well as cell shrinkage and the formation of apoptotic bodies were observed. FCM analysis showed hypodiploid peaks on histogram and the apoptotic rates at 5 ?g/ml concentration for 12, 24, 36, 48 and 72 h were (2.32?0.16)%, (3.01?0.35)%, (3.87? 0.43 )%, (6.73?0.58)% and (20.85?1.74)% respectively, which were all significantly higher than that of control group (1.07?0.13)%. Conclusion Tanshinone ⅡA can induce the apoptosis of human hepatoma cell line BEL 7402 in vitro , which may be related to the mechanism of growth inhibition of the human hepatoma cell line.

AIM: To study the effect of vascular endothelial growth factor(VEGF) on hematopoietic differentiation from mouse embryonic stem cells(ESC) in vitro.METHODS: ES-D3 was allowed to grow on mouse fetal fibroblast feeder layer,and then was developed into embryoid bodies(EB).EB cells were transferred into medium supplemented with different concentration of VEGF and VEGF+SCF for 1 week.Six groups,including.VEGF 5 ?g/L,VEGF 10 ?g/L,VEGF 20 ?g/L, VEGF 5 ?g/L+SCF,VEGF 10 ?g/L+SCF and VEGF 20 ?g/L+SCF,were designed.The group of spontaneous differentiation without cytokine(s) was used as control.Hematopoietic transcription factor GATA-2 and early hematopoietic differentiation genes(c-kit and ?-H1) were detected by RT-PCR.The content of CD34~+ cells in each group were measured by flow cytometry.The cells derived from ESC were incubated in semisolid methycellulose cultures.The numbers of total colony-forming units in culture(CFU-C) were counted by reverse microscope.RESULTS: ES-D3 grew and formed EB at day 4.VEGF had a stimulatory effect as a single factor on the expression of genes associated with early hematopoietic differentiation(GATA-2,c-kit and ?-H1),the generation of CD34~+ cells and CFU-C in EB.The effects of VEGF+SCF were the most potent in the experimental groups according to the percent of CD34~+ cells and the numbers of hematopoietic colonies.The most highest inducing efficacy was achieved in VEGF 20 ?g/L or VEGF 10 ?g/L combined with SCF.CONCLUSION: VEGF promotes the differentiation of ESC into hematopoietic cells in vitro.The strongest effect was achieved when VEGF was combined with SCF.

In order to study the effect of tanshinone II A on growth and apoptosis in human hepatoma cell line BEL-7402 in vitro, the human hepatoma cell line BEL-7402 was treated with tanshinone II A at various concentrations for 72 h. Growth suppression was evaluated by MTT assay; apoptosis-related alterations in morphology and biochemistry were ascertained under cytochemical staining (Hoechst 33258), transmission electron microscopy (TEM), and DNA agarose gel electrophoresis. Apoptotic rate was quantified by flow cytometry (FCM). The results showed that Tanshinone II A could inhibit the growth of hepatoma cells in a dose-dependent manner, with IC50 value being 6.28 micrograms/ml. After treatment with 1-10 micrograms/ml tanshinone II A for 72 h, BEL-7402 cells apoptosis with nuclear chromatin condensation and fragmentation as well as cell shrinkage and the formation of apoptotic bodies were observed. DNA ladder could be demonstrated on DNA electrophoresis. FCM analysis showed hypodiploid peaks on histogram, and the apoptotic rates at 5 micrograms/ml concentration for 12 h, 24 h, 36 h, 48 h and 72 h were (2.32 +/- 0.16)%, (3.01 +/- 0.35)%, (3.87 +/- 0.43)%, (6.73 +/- 0.58)% and (20.85 +/- 1.74)% respectively, which were all significantly higher than those in the control group (1.07 +/- 0.13)%. It is concluded that Tanshinone II A could induce human hepatoma cell line BEL-7402 apoptosis, which may be related to the mechanism of growth inhibition.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Division ; drug effects ; Cell Line, Tumor ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Humans ; Liver Neoplasms ; pathology ; Phenanthrenes ; pharmacology

Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.
Bacteriophages/genetics ; DNA, Complementary/*biosynthesis ; *DNA, Neoplasm/biosynthesis ; DNA, Recombinant/biosynthesis ; *Gene Library ; Genetic Vectors ; Leukemia, Promyelocytic, Acute/*genetics ; Leukemia, Promyelocytic, Acute/metabolism ; Leukemia, Promyelocytic, Acute/pathology ; RNA-Directed DNA Polymerase/metabolism ; Transcription, Genetic/genetics

Objective To investigate the clinical outcomes and complications of augmented antibiotic-loaded cement spacer in two-stage infected total hip arthroplasty with acetabular bone defect.Methods The periprosthetic infection (PJI) patients with acetabular bone defect were retrospectively reviewed from January 2007 to June 2016 in our hospital.A total of 26 patients (11 males and 15 females) were eligibly included in the present study.The mean age was 46.7 years old.The two-stage revision arthroplasty included implants removel,meticulous debridement,implantation of antibiotic-loaded cement spacer in firststage.After systemic therapy of antibiotics,the prosthesis was implanted in the second-stage.The supra-acetabular antibiotic cement shelf with screws was used to improve hip stability with acetabular wall defect.The handmade acetabular spacer was able to prevent femoral spacer ifto pelvis in patients with acetabular internal wall defect.The clinical outcomes and complications (spacer dislocation,spacer fracture and acetabular wear) were measured.Results The positive rate of bacteria culture was 80.8％ (21/26)and 57.7％ (15/26) patients were cultured with staphylococcus.The others were 2 fungus,2 Gram-positive rod,1 brucella,1 pseudomonas aeruginosa,1 escherichia coli,1 enterococcus faecalis,1 defective probiotics,1 serratiamarcescens and 1 Kocuriaroseus.Moreover,19.2％ (5/26) patients were mixed infection.There was one patient with spacer dislocation and two with spacer fracture.No patients were recurrent infection.Infection was controlled,and two-stage revision was successfully performed in 24 patients.Twenty-two patients were followed averaging 4.1 years (1-8) and the Harris Hip Score was significantly improved from 40.9± 14.0to 81.2± 11.2 at the final follow-up (P＜0.05).Conclusion The application of augmented antibiotic-loaded cement spacer has satisfactory clinical outcomes in PJI patients with acetabular bone defect.It can provide joint mobility and increase additional joint stability with decreased iatrogenic bone defect caused by acetabular wear.

Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.
Bacteriophages ; genetics ; DNA, Complementary ; biosynthesis ; DNA, Neoplasm ; biosynthesis ; DNA, Recombinant ; biosynthesis ; Gene Library ; Genetic Vectors ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; pathology ; RNA-Directed DNA Polymerase ; metabolism ; Transcription, Genetic ; genetics

Objective: To study the correlation of lifestyle characteristics with thyroid nodules in a population-based sample of centenarians in Hainan. Methods: The study was based on China Hainan Centenarian Cohort Study (CHCCS) conducted in 18 cities and counties in Hainan province from 2014 to 2016. A group of multidisciplinary team interviewed and examined local centenarians with structured questionnaires and ultrasonography procedures. A total of 918 centenarians were analyzed after excluding those who refused ultrasonographic examinations or had relevant missing data. Thyroids of centenarians were examined by 3-year experienced sonographer, details on lifestyle characteristics and dietary habits were collected by standard procedure. Results: Of the 918 centenarians, 683 (74.4%) had thyroid nodules under the ultrasonography procedures. The prevalence of thyroid nodules in different group of areca nut consumption varied significantly (P<0.05). In multivariate logistic regression model, the results showed female and areca nut intake were independently associated with thyroid nodules in centenarians (P<0.05). Conclusions The modifiable variables including areca nut intake as well as non-modifiable such as gender are independent determinants of thyroid nodules in centenarians. Further studies are warranted to explore and verify these associations in populations with different ranges of age.

Objective:To explore the clinical manifestations, diagnostic characteristics, treatment strategies and outcomes of patients with brucellaperi prosthetic joint infection (PJI).Methods:The medical records of 6 patients with brucella PJI in the First Medical Center of Chinese PLA General Hospital and the Third Hospital of Hebei Medical University from January 2010 to December 2018 were retrospectively analyzed, including 5 males and 1 female, aged 61.5±11.5 years (range 45-79 years) with body mass index 23.0±2.8 kg/m 2 (range 18.4-26.1 kg/m 2). Five cases lived in the countryside, 1 in the city. Four cases were farmers, while two cases were herder and unemployed. One case had contact history in the epidemic area, and 1 case had been in the slaughter industry. Three cases were with knee PJI, of which 1 patient underwent total knee arthroplasty due to knee joint villous nodular synovitis and 2 patients due to knee osteoarthritis. Three patients had hip PJI of which 1 patient underwent total hip arthroplasty due to spondylitis and hip ankylosis and 2 cases due to femoral head necrosis. Three cases were with acute PJI, while other 3 cases were with chronic PJI. Three cases showed fever, while 5 cases had local wounds swelling. A total of 4 cases were complicated with sinus tracts. Five cases had laboratory examinations on the day of admission of which 3 cases had elevated blood C-reactive protein (CRP) and 5 cases with increased erythrocyte sedimentation rate (ESR). Five cases were with increased blood interleukin-6 (IL-6), 2 cases with increased blood alanine transaminase (alanine transaminase, ALT). All cases had varying degrees of restricted movement of the affected joints. The normal range of motion of the hip joint was from 10° to 130°. The average range of motion of 3 patients with hip joint involvement was from 0° to 75°. The normal range of motion of the knee joint was 10°-135°. Three patients with knee joint involvement had an average range from -8° to 67°. One case showed loosening of the right hip prosthesis with infection and 1 case showed local soft tissue swelling. Other cases showed no obvious abnormalities in X-rays. Two patients who underwent frozen pathological examination during the operation had positive pathological neutrophilcounts. Four cases had positive Brucella culture in joint tissues or synovial fluid (1 case with mixed infection) and 2 cases had blood Brucella antibody positive. Results:Among the 3 cases of acute PJI, two of them were treated with debridement, antibiotics, irrigation and retention. One case was treated with two-stage revision. Among the 3 cases of chronic PJI, one was treated with two-stage revision and 2 were treated with one-stage revision. Brucella-specific antibiotics such as rifampicin and doxycycline were used in the antibiotic treatment with the course of antibiotics 3 to 12 weeks. At the time of discharge, the CRP and ESR dropped to the normal range (CRP 0-0.8 mg/dl, ESR 0-20 mm/1 h) in all cases except for the second case. Interleukin 6 was not tested in the sixth case before discharge. In the remaining 5 patients, the blood interleukin 6 fell to the normal range (0-5.9 pg/ml) in 2 cases, and the blood ALT was in the normal range (0-40 U/L) in 4 cases. The body temperature of the second case was 37.3 ℃, while the other cases dropped below 37.3 ℃. In the second case, fever occurred intermittently after surgery. Thus, the incisionwas reddened and swollen and exuded 2 months after the operation. The patient recovered after intravenous infusion of levofloxacin. Until the last follow-up, all patients had no recurrence of infection. Imaging examination comfirmed that the prosthesis was in good position.Conclusion:For patients with Brucella PJI, Brucella culture positive and Brucella antibody positive have specific diagnostic significance. Different surgical strategies will be adopt based on the patient's symptoms and the duration of infection. Surgery combined with Brucella specific antibiotic treatment can usually achieve satisfied therapeutic outcomes.

Objective:To explore the anti-epidemic preventions and perioperative management strategies of organ donation and liver transplantation during the pandemic period of novel coronavirus pneumonia (NCP) and summarize the experiences.Methods:On the basis of guidance of National Health Commission and Organ Transplantation Committee of Chinese Medical Association, anti-epidemic preventions and perioperative management strategies of organ donation and liver transplantation were adjusted under the background of NCP pandemic and the anti-epidemic preventions and treatment outcomes were evaluated. Eight organ donations and 7 liver transplantations were performed from February 4 to March 7, 2020. NCP infection screening results were negative in all pre-donation and pre-transplantation cases.Results:All donation operations and liver transplantations were successfully performed without postoperative complications. No NCP occurred during hospitalization period. Postoperative pulmonary infection occurred in 1 case (1/7) and the following novel coronavirus screening result was negative. Pulmonary inflammation became partially absorbed after antibacterial therapy.Conclusions:Through strict and effective anti-epidemic preventions and perioperative managements, organ donation and transplantation could be successfully performed during the pandemic period of NCP.