Induced pluripotent stem cells (iPSCs) can be propagated indefinitely, while maintaining the capacity to differentiate into all cell types in the body except for the extra-embryonic tissues. This iPSC technology not only represents a new way to use individual-specific stem cells for regenerative medicine but also constitutes a novel method to obtain large numbers of disease-specific cells for biomedical research. However, the low efficiency of reprogramming and genomic integration of oncogenes and viral vectors limit the potential application of iPSCs. Chemical-induced reprogramming offers a novel approach to generating iPSCs. In this study, a new combination of small-molecule compounds (SMs) (sodium butyrate, A-83-01, CHIR99021, Y-27632) under conditions of transient folate deprivation was used to generate iPSC. It was found that transient folate deprivation combined with SMs was sufficient to permit reprogramming from mouse embryonic fibroblasts (MEFs) in the presence of transcription factors, Oct4 and Klf4, within 25 days, replacing Sox2 and c-Myc, and accelerated the generation of mouse iPSCs. The resulting cell lines resembled mouse embryonic stem (ES) cells with respect to proliferation rate, morphology, pluripotency-associated markers and gene expressions. Deprivation of folic acid, combined with treating MEFs with SMs, can improve the inducing efficiency of iPSCs and reduce their carcinogenicity and the use of exogenous reprogramming factors.

BACKGROUNDStreptococcus mutans (S. mutans) is the prime pathogen of dental caries. There are few reports that studied the relationship between S. mutans, bacteria and dental caries in permanent teeth when compared to those in primary teeth. This study aimed to detect S. mutans and bacteria of dental caries and non-caries groups in permanent teeth from a north China population by real-time polymerase chain reaction (PCR) and compare the relationship between the number of these bacteria and the prevalence of dental caries in permanent teeth.

METHODSHuman saliva samples were collected from 142 subjects with permanent teeth. According to their dental tooth (DT), 142 subjects were divided into a dental caries group (DT ≥ 1) and a non-caries group (DT = 0). With specific primers for S. mutans and 16S rRNA, the total number of S. mutans and total bacteria of 142 saliva samples were detected by real-time PCR and statistically analyzed.

RESULTSThere was no significant difference between the detection rates of S. mutans (P = 0.118) and medians of S. mutans (P = 0.115). The ratio of S. mutans to total bacteria in people with dental caries was significantly higher than in those without caries (P < 0.001), but the total number of bacteria in people with dental caries was significantly lower than in those without caries (P < 0.001).

CONCLUSIONSS. mutans had different effects on caries in the permanent teeth of several individuals from a north China population. The ratios of S. mutans to total bacteria in saliva detected by real-time PCR with Sm479F/R and 16S RNA primers were closely associated with the prevalence of dental caries in the same population. These assays may be useful for the assessment of an individual's risk of dental caries.

Adolescent ; Adult ; Aged ; Bacteria ; isolation & purification ; Dental Caries ; microbiology ; Female ; Humans ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Saliva ; microbiology ; Sensitivity and Specificity ; Streptococcus mutans ; isolation & purification ; Tooth ; microbiology

OBJECTIVETo establish a quantity detection method of Streptococcus mutans (Sm) and bacteria and compare the relationship between the number of these bacteria and the prevalence of dental caries in different people.

METHODSWith specific primers for a unique sequence in a 14 kb HaeIII restriction fragment consistently presenting during detecting Sm by chromosomal DNA fingerprints, the total number of Sm and bacteria of 99 saliva samples were detected by real-time polymerase chain reaction (PCR) and statistically analyzed.

RESULTSThe primers were specific for Sm and the minimum detectable level by real-time PCR was 0.1 microg/L. The total number of bacteria in the dental caries and people without caries was 51.4 x 10(8) cell copies/L and 221.6 x 10(8) cell copies/L respectively, in which the ratio of Sm to bacteria was 0.0193 and 0.0059 respectively. The differences were significantly different between the people with dental caries and those without caries in the total number of bacteria and the ratio of Sm to bacteria.

CONCLUSIONSThe primers can be used to detect the Sm by real-time PCR. The ratio of Sm to bacteria was closely associated with the prevalence of dental caries.

Adult ; DNA Primers ; DNA, Bacterial ; analysis ; Dental Caries ; epidemiology ; microbiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Real-Time Polymerase Chain Reaction ; Saliva ; microbiology ; Streptococcus mutans ; genetics ; isolation & purification

Objective To investigate the infection and genotype of hantaviruses in rodents from Wuhan area,Hubei province.Methods Rodents were trapped in fields and residential areas of Xinzhou and Jiangxia districts of Wuhan in autumn and winter seasons,from 2000 to 2003 and from 2009 to 2011.Trapped rodents were identified,and hantavirus antigens were detected in the lung tissues with indirect immunofluorescence assay (IFA).Partial S segment sequences were amplified with RT-PCR in hantavirus antigen positive samples and then sequenced.Phylogenetic tree was constructed to analyze the genetic characteristics of hantaviruses.Results From 2000 to 2003,437 rodents were trapped,with 24 (5.49％) lung tissues showed hantavirus antigen positive.From 2009 to 2011,173 rodents were trapped and 7 (4.05％) were hantavirus antigen positive.Rattus norvegicus were the dominant species of rodents.Partial S segment sequences were amplified from 22 samples with Hantaan and Seoul viruses specific primers and sequenced.Partial S segments of Seoul viruses (nucleotide 588-1147) were amplified from 17 rodents (13 R.norvegicus and 4 Apodemus agrarius).Seven of these sequences belonged to 3 genetic lineage,while two novel genetic lineages were formed by 9 and 1 sequences,respectively.Partial S segments of Hantaan viruses (nucleotide 615-1141 ) were amplified from 5 A.agrarius.One of these sequences belonged to 7 genetic lineages,and 4 sequences formed one novel genetic subtype.Conclusion Hantaan and Seoul viruses co-circulated in Wuhan area.Hubei province.Novel genetic lineages were identified in this study and Seoul virus might have caused spillover infection in A.agrarius.

Induced pluripotent stem cells (iPSCs) can be propagated indefinitely, while maintaining the capacity to differentiate into all cell types in the body except for the extra-embryonic tissues. This iPSC technology not only represents a new way to use individual-specific stem cells for regenerative medicine but also constitutes a novel method to obtain large numbers of disease-specific cells for biomedical research. However, the low efficiency of reprogramming and genomic integration of oncogenes and viral vectors limit the potential application of iPSCs. Chemical-induced reprogramming offers a novel approach to generating iPSCs. In this study, a new combination of small-molecule compounds (SMs) (sodium butyrate, A-83-01, CHIR99021, Y-27632) under conditions of transient folate deprivation was used to generate iPSC. It was found that transient folate deprivation combined with SMs was sufficient to permit reprogramming from mouse embryonic fibroblasts (MEFs) in the presence of transcription factors, Oct4 and Klf4, within 25 days, replacing Sox2 and c-Myc, and accelerated the generation of mouse iPSCs. The resulting cell lines resembled mouse embryonic stem (ES) cells with respect to proliferation rate, morphology, pluripotency-associated markers and gene expressions. Deprivation of folic acid, combined with treating MEFs with SMs, can improve the inducing efficiency of iPSCs and reduce their carcinogenicity and the use of exogenous reprogramming factors.
Amides ; pharmacology ; Animals ; Butyric Acid ; pharmacology ; Cell Differentiation ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Extraembryonic Membranes ; cytology ; drug effects ; Folic Acid ; pharmacology ; Induced Pluripotent Stem Cells ; cytology ; drug effects ; Kruppel-Like Transcription Factors ; metabolism ; Mice ; Octamer Transcription Factor-3 ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism ; Pyrazoles ; pharmacology ; Pyridines ; pharmacology ; Pyrimidines ; pharmacology ; SOXB1 Transcription Factors ; metabolism ; Thiocarbamates ; pharmacology ; Thiosemicarbazones

BACKGROUNDLaparoscopic Heller cardiomyotomy and Dor fundoplication is the surgical procedure of choice for esophageal achalasia. The aim of this study was to investigate the clinical outcome of laparoscopic Heller-Dor procedure in our initial series of 25 patients with achalasia.

METHODSBetween October 2003 and January 2006, a total of 25 patients with achalasia underwent laparoscopic Heller-Dor operation. Among them, 9 were male and 16 were female with an average age of (41.5 +/- 5.1) years (21-66). All the patients received upper gastrointestinal series (barium swallow), esophagogastroscopy, esophageal manometry to exclude esophageal carcinoma and to confirm the diagnosis, and 21 patients also had 24-hour ambulatory pH studies. All the patients were operated by laparoscopic modified Heller's myotomy with Dor fundoplication. In addition, 2 of them had combined laparoscopic cholecystectomy + excision of hepatic hemangioma and laparoscopic cholecystectomy, respectively.

RESULTSThe average operating time was (110.6 +/- 12.9) minutes (range, 60-180), operative blood loss averaged (18.6 +/- 7.1) ml (5-50), the median time to oral feeding was (1.6 +/- 0.4) days (1-4) and the median hospital stay was (12.6 +/- 1.2) days (10-20). There was no conversion to open surgery. Intraoperative mucosal perforation was encountered in six patients and was repaired in all of them by laparoscopic suture. All the patients had an uneventful recovery without postoperative complication. After a median follow-up of (10.6 +/- 7.2) months (1-27), 24 patients were asymptomatic and 1 had mild postoperative dysphagia.

CONCLUSIONSLaparoscopic Heller-Dor operation had the advantages of reduced compromise of the cardiopulmonary function, with less disruption of the supporting structures (phrenoesophageal membrane) of the antireflux mechanism, requiring simpler general anesthesia and providing excellent exposure permitting an easy fundoplication, less pain and reduced morbidity, shorter hospitalization and faster convalescence.

Adult ; Aged ; Digestive System Surgical Procedures ; methods ; Esophageal Achalasia ; surgery ; Esophagus ; surgery ; Female ; Follow-Up Studies ; Fundoplication ; methods ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods

OBJECTIVETo explore the effect of extracellular signal regulated kinase 1/2 (ERK1/2) on edaravone (EDA)-triggered protection against myocardial toxicity induced by isoprenaline (ISO) in H9c2 myocardial cells (H9c2 cells).

METHODSH9c2 cells were exposed to ISO at different concentrations to establish a cardiac toxicity model induced by persistent excitation of β1 receptor. EDA was added before ISO as a pretreatment. PD-98059, an ERK1/2 inhibitor, was administered 1 h prior to EDA to inhibit the phosphorylation of ERK1/2. Cell viability was measured using cell counter kit (CCK-8). The expressions of p-ERK1/2 and t-ERK1/2 were tested by Western blotting. Mitochondrial membrane potential (MMP) was detected by Rhodamine123 (Rh123) staining and photofluorography.

RESULTSExposure of H9c2 cells to 80 µmol/L ISO for 24 h down-regulated ERK1/2 phosphorylation and repressed MMP. Pretreatment with 10-40 µmol/L EDA for 1 h inhibited ISO-induced myocardial toxicity and pretreatment of 40 µmol/L EDA partially rescued ERK1/2 phosphorylation and MMP level. PD-98059 abolished cardiac protection of EDA, leading to myocardial toxicity and MMP loss.

CONCLUSIONEDA can protect H9c2 cells against myocardial injury induced by ISO by suppressing ISO-triggered inhibition of ERK1/2 activation.

Animals ; Antipyrine ; analogs & derivatives ; pharmacology ; Cell Line ; Flavonoids ; pharmacology ; Isoproterenol ; toxicity ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; Phosphorylation ; Rats

Contagious ecthyma (also known as orf) is an acute skin zoonosis caused by orf virus (ORFV), which affects sheep, goats and humans. As one of the typical species of the Parapoxvirus genus of the Poxviridae family, orf virus has distinctive and unique characteristics of these species. A range of immuno-modulatory/pathogenesis -related genes acquired by virus that function is to limit (at least transiently) the effectiveness of host immunity during its evolution. This review is aimed to describe the latest progress on the molecular characteristics of ORFV, and upon which we analyzed molecular mechanism of the immune escape designed and a set of strategies developed for ORFV to effective against immune clearance of the host. Known as an essential component in evolutionary system, host is regulated by ORFV for using in population evolution. By the ORFV evolutional immune regulation components and its effect approach, we can understand the viral biological characteristics of ORFV, and it is helpful for us to further study the counter-measures of this disease.
Animals ; Ecthyma, Contagious ; immunology ; virology ; Gene Expression Regulation, Viral ; Immune Evasion ; Orf virus ; genetics ; immunology ; Viral Proteins ; genetics ; immunology

BACKGROUNDMicroalbuminuria (MAU) is a key component of metabolic syndrome (MetS) and is an early sign of diabetic nephropathy as well. Although routine Western medicine treatments are given to MetS patients to control high blood pressure, hyperglycemia and dyslipidemia, some patients still experience progressive renal lesions and it is necessary to modify and improve the treatment strategy for MetS patients.

OBJECTIVETo investigate the efficacy of Yiqi Huaju Qingli Herb Formula, a compound traditional Chinese herbal medicine, in MetS patients with MAU when it is combined with routine Western medicine treatment.

DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONSSixty patients with MetS were randomized into the Chinese herbal formula group (CHF, Yiqi Huaju Qingli formula treatment in combination with Western medicine) and control group (placebo in combination with Western medicine). All treatments were administered for 12 weeks.

MAIN OUTCOME MEASURESUrinary microalbumin (MA), urinary albumin-to-creatinine ratio (UACR), 24-hour total urine protein (24-hTP), body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), fasting plasma glucose (FPG), 2-hour postprandial plasma glucose (2-hPPG), glycosylated hemoglobin (HbA1c), homeostasis model assessment for insulin resistance (HOMA-IR), blood lipid profile and blood pressure were observed.

RESULTSCompared with the control group, CHF treatment significantly decreased BMI (P<0.05), WC (P<0.01) and WHR (P<0.01). Both groups had significant decreases in FPG, 2-hPPG, HbA1c, HOMA-IR, MA, and UACR, with CHF treatment showing better effects on these parameters compared with the control treatment (P<0.05). Both treatments significantly reduced the levels of total cholesterol, low-density lipoprotein cholesterol and triacylglycerol (TAG), and a greater reduction in TAG was observed with CHF treatment (P<0.05). The level of high-density lipoprotein cholesterol did not change in the control group after treatment (P>0.05), whereas it significantly increased with CHF treatment (P<0.01). Compared with before the treatment, significant decreases in systolic blood pressure, diastolic blood pressure and mean arterial blood pressure were observed in both groups (P<0.01). However, there was no significant difference between the two groups (P>0.05).

CONCLUSIONCombined treatment of Yiqi Huaju Qingli Formula and Western medicine significantly alleviated MAU, which may correlate with the improvement of insulin sensitivity and glucose and lipid metabolism. TRIAL REGISTRATION IDENTIFIER: This trial was registered in the Chinese Clinical Trial Registry with the identifier ChiCTR-TRC-11001633.

Adolescent ; Adult ; Aged ; Albuminuria ; drug therapy ; etiology ; metabolism ; Blood Glucose ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Lipids ; blood ; Male ; Metabolic Syndrome ; complications ; drug therapy ; metabolism ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo study the situation of tuberculosis (TB) infection among the employees of the anti-TB institutions in Henan.

METHODSCross-sectional study was adopted the employees working in all municipal-level- anti-TB institutions and 40 anti-TB institutions at county-level selected randomly from 109 counties of the province were regarded as surveyed objects. Tuberculin skin test (TST) was used to test the infection with PPD.

RESULTS2153 employees accepting the TST and the positive rate was 60.6%, of which the positive rate was 66.1% among healthcare workers. Among the employees and healthcare workers, the positive rates of TST adjusted by the stratum weights between municipal-level and county-level institutions were 57.3% and 62.8% respectively with Chi-square test the analysis of multivariate logistic vegression, both positive rate and strong positive rate among healthcare workers, the employees older than 30 years of age and working in municipal-level institutions were significantly higher than those among non-healthcare workers, the employees younger than 30 years old and working in county-level institutions, respectively. There were not significant differences of positive and strong positive rates between employees with and without BCG-history, or between male employees and female employees.

CONCLUSIONProgram on TB infection control in anti-TB institutions of Henan were weak and the employees especiolly healthcare workers had a high vocational exposure.

Adult ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Hospitals, Chronic Disease ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Occupational Diseases ; epidemiology ; Personnel, Hospital ; statistics & numerical data ; Population Surveillance ; Prevalence ; Tuberculin Test ; Tuberculosis ; epidemiology