BACKGROUND:Reprogramming somatic cells to generate pluripotent stem cells has a wide application in biomedical research. OBJECTIVE:To analyze the effect of different molecular weight proteins in chicken egg-white extract to elevate expression of pluripotent genes Oct-3/4 and Nanog in 293T cells. METHODS:The extracts of chicken egg-white were separated into more than 3 ku and less than 3 ku ingredients to be used for co-culture with 293T cells. There were four groups, 1×105 293T cells per wel , total 500μL. In the control group, 500μL culture medium was added;in the other three groups, 500μL chicken egg-white extract, more than 3 ku and less than 3 ku ingredients were respectively added. Quantitative PCR was used to determine the relative expression levels of pluripotent genes Nanog and Oct-3/4 in 293T cells. RESULTS AND CONCLUSION:By using co-culture method, more than 3 ku ingredients have a role to increase the expression of pluripotent genes Oct-3/4 and Nanog, but less than 3 ku ingredients cannot elevate the expression of pluripotent genes. This indicates that the ingredient of chicken egg-white extract to elevate the expression of pluripotent genes is more than 3 ku proteins.

BACKGROUND:As the high proliferation and low apoptosis of the bone marrow in polycythemia vera patients, hematopoietic stem cels transplanted into NOD/SCID mice can differentiate into erythroid cels, but whether hematopoietic stem cels transplantation could improve the hematopoietic function of aplastic anemia mice is not yet reported. OBJECTIVE:To investigate whether transplantation of bone marrow mononuclear cels with JAK2V617F mutation from polycythemia vera patients can influence hematopoietic reconstruction in aplastic anemia mice. METHODS:Severe aplastic anemia mouse models were established by using recombinant human interferon-γplus busulfan, and then, these mouse models were randomly divided into experimental group (n=10) and control group (n=10). Bone marrow mononuclear cels isolated from polycythemia vera patients with positive JAK2V617F mutation were transplanted into the mice in the experimental group via tail vein at 5 days after drug withdrawal.The same volume of normal saline was administered to the control group. Routine peripheral blood test, morphology of bone marrow cels, bone marrow biopsy, and percentage of CD45+ cels in the peripheral blood and marrow were determined at 14 days after transplantation. RESULTS AND CONCLUSION: At 14 days after transplantation, pancytopenia occurred in the experimental group, bone marrow smears showed scattered lymphocytes and hematopoietic progenitors, and bone marrow biopsy presented that hematopoietic tissues were reduced and a smal amount of granulocyte cels and erythroblasts could be seen, but megakaryocytes were rare. In contrast to the control group, there was no improvement in the hematopoietic function of mice in the experimental group. CD45+ cels were detectable in the peripheral blood and bone marrow in the experimental group, but not in the control group; and a higher percentage of CD45+ cels was measured in the bone marrow than in the peripheral blood of experimental group mice. Experimental findings indicate that bone marrow mononuclear cels from polycythemia vera patients with positive JAK2V617F mutation can be engrafted into aplastic anemia mice, but cannot improve the hematopoietic function of mice.

Objective: To explore the effectiveness of preventive treatment for latent tuberculosis infection (LTBI) patients, so as to provide reference for the management and preventive treatment of clustered epidemic in schools. Methods: Data came from the school tuberculosis outbreak of a boarding high school in Kaizhou District, Chongqing, which occurred between June 2017 and March 2018 among 2016 grade high school teachers and students for investigation and analysis. The total incidence, LTBI patients, and the incidence after preventive treatment for 5 years were followed up. Results: A total of 34 cases of pulmonary tuberculosis from June 2017 to March 2018. A total of 1 357 individuals were screened for 6 concentrated contact screenings, with a confirmed tuberculosis rate of 2.43%, a tuberculosis skin test (TST) positive rate of 27.41%, and a strong TST positive rate of 7.39%. Among them, the confirmed tuberculosis rate and TST positive rate in the first case class were much higher than those in other classes, with statistically significant differences ( χ 2=286.30, 98.59, P ＜0.01). 88 cases of LTBI were found, with 31 cases receiving preventive treatment (35.23%), of which 28 completed preventive treatment (90.32%). After five years of follow-up, 73 cases of pulmonary tuberculosis were diagnosed in 2016 by the school senior high school, with a incidence rate of 0.98/10 2 (person/person years). Fifteen of the 88 LTBI patients were diagnosed with pulmonary tuberculosis, and the incidence rate was 3.33/10 2 (person/person years). The incidence rate of the preventive treatment group was 0.7/10 2 (person/person years)lower than that of the medical observation group 4.5/10 2 (person/person years), with a statistically significant difference ( χ 2=4.31, P ＜0.05). Conclusion The classes with higher TST positive rate and strong positive rate have higher incidence rate. Improving the preventive treatment rate of LTBI patients can effectively reduce the incidence rate of tuberculosis.

OBJECTIVETo evaluate the ability of a polycaprolactone/polylactic acid (PCL/PLA) membrane to inhibit epidural scar adhesion after laminectomy, and observe the responsive changes of the pain media in the spinal cord.

METHODSL(1), L(3) laminectomies were performed on 96 Wistar rats. The rats were divided into 3 groups: None-implant Control Group (NC), Autologous free fat graft group (AFFG) and PCL/PLA membrane group (PCL/PLAm). The rats were killed at 1, 3, 6, and 12 weeks postoperatively. Epidural scar formation and adhesion were observed grossly and histologically. Reverse transcription polymerase chain reaction (RT-PCR) were used to analyses the expression of Transforming growth factor beta (TGF-beta) in the epidural scar. Immunohistochemistry stain and RT-PCR were performed to evaluate the expression of the substance P and the c-fos gene in the relevant spinal cord, and the results were analyzed statistically.

RESULTSGross evaluation and histological evaluation showed that in the NC lamina defect site had much scar tissue and had wide and tight adhesions to the dura; in the AFFG, with the fat degrading gradually, the adhesions were increased; whereas in the PCL/PLAm group, there were slightly adhesions to the dura. RT-PCR showed that the expression of the TGF-beta was much less in the PCL/PLAm group than in the NC group. The insertion of the PCL/PLA membrane and the fat patch reduced the expression of the substance P and the c-fos gene in the spinal cord.

CONCLUSIONThe insertion of the PCL/PLA membrane reduces scar formation and separates fibrosis tissue from the dura, the results indicate that PCL/PLA membrane is an effective way of reducing peridural scar formation and preventing the failed back surgery syndrome.

Animals ; Biocompatible Materials ; Cicatrix ; prevention & control ; Female ; Lactic Acid ; Laminectomy ; adverse effects ; Membranes, Artificial ; Polyesters ; Polymers ; Postoperative Complications ; prevention & control ; Prosthesis Implantation ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Rats ; Rats, Wistar ; Spinal Cord ; metabolism ; Spinal Diseases ; prevention & control ; Substance P ; biosynthesis ; Tissue Adhesions ; prevention & control

BACKGROUNDPrevious studies show that sleep-related breathing disorder (SRBD) is common in patients with heart failure (HF) and is associated with increased mortality. This study aimed to determine whether there was significant difference of subjective daytime sleepiness between HF patients with and without SRBD.

METHODSWe enrolled, prospectively, 195 consecutive HF patients with left ventricular ejection fractions (LVEF) < or = 45% and all subjects underwent polysomnography to measure the sleep structure between 2005 and 2008. Patients were then assigned to those with SRBD including obstructive and central sleep apnea (apnea-hypopnea index (AHI) > or = 5/hour of sleep) and those without SRBD (AHI < 5/hour) according to the sleep study. The subjective sleepiness was assessed with Epworth sleepiness scale (ESS).

RESULTSAmong 195 HF patients, the prevalence of obstructive sleep apnea (OSA) was 53% and of central sleep apnea (CSA) was 27%. There was no significant difference of ESS scores between patients without SRBD (NSA) and with SRBD (NSA vs OSA: 6.7 +/- 0.6 vs 7.6 +/- 0.4, P = 0.105 and NSA vs CSA: 6.7 +/- 0.6 vs 7.4 +/- 0.5, P = 0.235, respectively), indicating that SRBD patients had no more subjective daytime sleepiness. Compared with NSA, patients with SRBD had increased arousal index (ArI) (NSA vs OSA: 14.1 +/- 1.4 vs 26.3 +/- 1.5, P < 0.001 and NSA vs CSA: 14.1 +/- 1.4 vs 31.3 +/- 3.5, P < 0.001, respectively), more awake number after sleep onset (NSA vs OSA: 19.2 +/- 1.5 vs 26.2 +/- 1.4, P = 0.01 and NSA vs CSA: 19.2 +/- 1.5 vs 36.9 +/- 4.4, P < 0.001, respectively), and reduced proportion of slow-wave sleep (SWS) (NSA vs OSA: 13.8 +/- 1.7 vs 9.3 +/- 0.7, P = 0.024 and NSA vs CSA: 13.8 +/- 1.7 vs 8.9 +/- 0.9, P = 0.024, respectively).

CONCLUSIONSOSA and CSA remain common in patients with HF on optimal contemporary therapy. Patients with both HF and SRBD have no significant subjective daytime sleepiness compared with patients without SRBD, despite of significantly increased awake number, arousal and decreased proportion of deep sleep stages. It is not a credible way and means to exclude SRBD in patients with HF according to the absence of subjective daytime sleepiness.

Adult ; Aged ; Aged, 80 and over ; Female ; Heart Failure ; physiopathology ; Humans ; Male ; Middle Aged ; Polysomnography ; Sleep Apnea Syndromes ; epidemiology ; etiology

BACKGROUNDAnkylosing spondylitis (AS) is the most common rheumatic condition that is slowly progressive and predominantly affects adolescents. Pathological bone formation associated with AS is an important cause of disability. The aim of the study was to investigate the possible involvement of the genes related to endochondral ossification and ectopia ossification in genetic susceptibility to AS in a Chinese Han population.

METHODSSixty-eight single nucleotide polymorphisms (SNPs) from 13 genes were genotyped in discovery cohorts including 300 AS patients and 180 healthy controls. The rs10019009 in dentin matrix protein 1 (DMP1) gene shown as association with AS after multiple testing corrections in discovery cohorts was replicated in a validation independent cohort of 620 AS patients and 683 healthy controls. The rs10019009 was assessed with bioinformatics including phylogenetic context, F-SNP and FastSNP functional predictions, secondary structure prediction, and molecular modeling. We performed a functional analysis of rs10019009 via reverse transcription-polymerase chain reaction, alkaline phosphatase (ALP) activity in human osteosarcoma U 2 OS cells.

RESULTSInterestingly, the SNP rs10019009 was associated with AS in both the discovery cohort (P = 0.0012) and validation cohort (P = 0.0349), as well as overall (P = 0.0004) in genetic case-control association analysis. After a multivariate logistic regression analysis, the effect of this genetic variant was observed to be independent of linkage disequilibrium. Via bioinformatics analysis, it was found that the amino acid change of the rs10019009 led to changes of SNP function, secondary structure, tertiary conformation, and splice mode. Finally, functional analysis of rs10019009 in U 2 OS cells demonstrated that the risk T allele of the rs10019009 increased enzymatic activity of ALP, compared to that of the nonrisk allele (P = 0.0080).

CONCLUSIONSThese results suggested that the DMP1 gene seems to be involved in genetic predisposition to AS, which may contribute to the ectopic mineralization or ossification in AS. In addition, DMP1 gene may be a promising intervention target for AS in the future.

Adult ; China ; ethnology ; Extracellular Matrix Proteins ; chemistry ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Logistic Models ; Male ; Phosphoproteins ; chemistry ; genetics ; Polymorphism, Single Nucleotide ; Spondylitis, Ankylosing ; etiology ; genetics

OBJECTIVETo determine the incidence and risk factors of HCV infection among heroin addicts who were receiving methadone maintenance treatment(MMT)in Dehong prefecture, Yunnan province.

METHODSAll heroin addicts who were HCV negative at the initiation of MMT in June 2005 through March 2012, in Dehong prefecture, were included in this cohort analysis. HCV incidence was calculated and related risk factors determined by using Cox proportional hazard regression model.

RESULTSA total of 2390 MMT clinic attendants were qualified for this cohort study by March 2012. 731(30.6%) of them had never received any follow-up HCV testing so were recognized as loss to follow-up. The other 1659 (69.4%) participants had received at least one follow-up HCV testing and were observed for a total of 3509.12 person-years(py). During this period 99 new HCV infections or HCV sero-converters were identified. The overall HCV incidence was 2.82/100 py and was 3.62/100 py for 2006, 5.36/100 py for 2007, 6.71/100 py for 2008, 2.56/100 py for 2009, 1.90/100 py for 2010, and 0.44/100 py for 2011, respectively. Results from multiple regression analysis, using Cox proportional hazard model, indicated that after controlling for confounding variables, those who were unemployed, being injecting drug users(IDUs)or HIV positive at entry into the MMT program were more likely to be newly infected with HCV or HCV sero-converted during the follow-up period than those who were peasants, non-IDUs or HIV negative at entry into the MMT program(HR = 2.02, 95% CI:1.18-3.48; HR = 9.05, 95% CI:5.49-14.93; HR = 2.12, 95% CI: 1.37-3.56), respectively.

CONCLUSIONThe incidence of HCV infection among MMT clinic attendants was decreasing since 2009 in Dehong prefecture. Those who were unemployed, injecting drug users and HIV positive were at higher risk of HCV infection.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Cohort Studies ; Drug Users ; Female ; Hepatitis C ; epidemiology ; Humans ; Incidence ; Male ; Methadone ; therapeutic use ; Risk Factors ; Substance-Related Disorders ; drug therapy ; epidemiology ; virology

OBJECTIVETo report the experience of the surgical treatment of congenital heart diseases (CHD) in pediatric patients with body weight less than 10 kg.

METHODSBetween January 2000 and December 2007, 105 children with CHD, aged 2 months to 3 years and weighing between 3.5 to 10 kg, underwent surgical treatment. Of the 105 patients, 56 were concomitant with moderate to severe pulmonary hypertension or repeated pulmonary infections, and 35 with complex cardiac abnormalities. Operations were performed through median sternotomy with moderate hypothermic cardiopulmonary bypass in 88 cases, with normothermic extracorporeal circulation on beating heart in 14 cases, and with deep hypothermic circulatory arrest in 3 cases.

RESULTSThere were 5 early deaths (4.8%). During a follow-up of 2 month to 3 year, 97 survivors with corrective procedure had no late mortality or complications with NYHA class I of cardiac function, excepting 2 cases with little residual shunt. Three survivors with palliative procedure enjoyed higher quality of life.

CONCLUSIONSThe surgical treatment of CHD in pediatric patients with body weight less than 10 kg seems to be feasible and safe, with satisfactory early and long-term results.

Body Weight ; Cardiac Surgical Procedures ; adverse effects ; methods ; Child, Preschool ; Female ; Heart Defects, Congenital ; mortality ; surgery ; Humans ; Infant ; Male

Objective To determine the incidence and risk factors of HIV infection among heroin addicts receiving methadone maintenance treatment(MMT)in Dehong prefecture,Yunnan province.Methods All heroin addicts who were HIV negative at the initiation of MMT in June 2005 and through June 2011,in Dehong prefecture were included in the cohort analysis.HIV incidence was calculated and related risk factors determined by using Cox proportional hazard regression model.Results A total of 3154 MMT clinic attendants were qualified for this cohort study.By June 2011,1023(32.4％)of them had never received any follow-up HIV testing so were thus referred as loss to follow-up.The other 2131(67.6％)members had received at least one follow-up HIV testing and were observed for a total of 4615.86 person-years.During the period,22 new HIV infections or seroconverters were identified,making the overall HIV incidence as 0.48/100 person-years.The HIV incidence was higher among those who were unemployed,never married,self-reported being injecting drug users(IDUs)and HCV positive at entry into the MMT program.None of those who were always negative on follow-up-urine-testing of morphine was discovered as HIV newly infected during the follow-up period.Data from multiple regression analysis under Cox proportional hazard model indicated that after controlling for confounding variables,non-IDUs at the entry point for the MMT program,were less likely to be HIV newly-infected or seroconverted than IDUs(HR=0.29,95％CI:0.11-0.76).Conclusion MMT prograqm in Dehong prefecture was demonstrated to be fairly effective in reducing HIV transmission through drug use.Those HIV negative attendants at the MMT clinic who were IDUs or keep using drugs during the treatment,were at higher risk of HIV seroconvertion.More efforts were needed to improve the follow-up and HIV testing programs for the MMT clinic attendants.

OBJECTIVEIn mid-July 2005, five patients presented with septic shock to a hospital in Ziyang city in Sichuan, China, to identify the etiology of the unknown reason disease, an epidemiological, clinical, and laboratory study were conducted.

METHODSAn enhanced surveillance program were established in Sichuan, the following activities were introduced: active case finding in Sichuan of (a) laboratory diagnosed Streptococcus suis infection and (b) clinically diagnosed probable cases with exposure history; supplemented by (c) monitoring reports on meningococcal meningitis. Streptococcus suis serotype 2 infection was confirmed by culture and biochemical reactions, followed by sequencing for specific genes for serotype and virulence factors.

RESULTSFrom June 10 to August 21, 2005, 68 laboratory confirmed cases of human Streptococcus suis infections were reported. All were villagers who gave a history of direct exposure to deceased or sick pigs in their backyards where slaughtering was performed. Twenty six (38%) presented with toxic shock syndrome of which 15 (58%) died. Other presentations were septicaemia or meningitis. All isolates were tested positive for genes for tuf, species-specific 16S rRNA, cps2J, mrp, ef and sly. There were 136 clinically diagnosed probable cases with similar exposure history but incomplete laboratory investigations.

CONCLUSIONAn outbreak of human Streptococcus suis serotype 2 infections occurred in villagers after direct exposure to deceased or sick pigs in Sichuan. Prohibition of slaughtering in backyards brought the outbreak to a halt. A virulent strain of the bacteria is speculated to be in circulation, and is responsible for the unusual presentation of toxic shock syndrome with high case fatality.

Animals ; Bacteremia ; epidemiology ; microbiology ; China ; epidemiology ; Disease Outbreaks ; Humans ; Meningitis, Bacterial ; epidemiology ; microbiology ; Shock, Septic ; epidemiology ; microbiology ; Streptococcal Infections ; epidemiology ; microbiology ; veterinary ; Streptococcus suis ; isolation & purification ; Swine ; Swine Diseases ; microbiology