This study aimed to construct full-length cDNA clones of the Japanese encephalitis virus (JEV). SA14-14-2 strain and discuss the feasibility of constructing chimeric viruses for exogenous gene expression based on the JEV genetic skeleton. Long-fragment RT-PCR techniques were applied to amplify JEV cD-NAs, and two amplified fragments with corresponding restriction endonuclease sites at both ends were cloned into the pACYC184 vector sequentially. Using standard molecular techniques, the enhanced green fluorescent protein (EGFP) gene was inserted into the 3' non-coding region of JEV as a reporter gene. After in vitro transcription and transfection procedures, wild-type JEV and chimeric JEV that expressed the EGFP as the reporter gene were successfully rescued. The recovered viruses were characterized by RT-PCR, plaque assays, and direct fluorescence microscopy. After six serial passage generations, the stability of the recovered viruses were studied in terms of virus growth characteristics and structural gene expression. The results showed that cDNA clones of rJEV and rJEV-EGFP were successfully constructed and rescued in BHK-21 cells after in vitro transcription and transfection. Each generation of the recovered viruses was stable and the chimeric virus rJEV-EGFP could stably express EGFP. The findings of this study indicate that both rJEV and rJEV-EGFP could be constructed and rescued in BHK-21 cells, and the JEV SA14-14-2 strain could be obtained as a viral vector to express foreign genes.
Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Encephalitis Virus, Japanese ; genetics ; metabolism ; Encephalitis, Japanese ; virology ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Vaccines, Attenuated ; genetics ; metabolism ; Viral Vaccines ; genetics ; metabolism

Objective To explore hemodynamic parameter changes in pulmonary arterial hypertension(PAH)treated by transplantation of bone marrow mesenchymal stem cells(MMSCs)in the experimental rats.Methods MMSCs cells were collected from bone marrow of Sprague-Dawleye(SD)rat's femoral and tibial bones,cultured and passaged in vitro,then stained by Hoechst 33342 fluorescence dye stuff.Ninety healthy male SD rats were randomly divided into 3 groups(n=30):normal control group(group N),MMSCs transplanted group(group M),PAH model group(group H).The rats in the two latter groups were given a single subcutaneous crotaline(50 mg/kg)to establish the model of PAH.The rats of group N were injected respectively a single subcutaneous 9 g/L saline water(6 mL/kg).After 21 days,5?109 L-1 MMSCs cultured in 1 mL phosphate-buffered saline were infused into the rats respectively in group M by sublingual vein and 1 mL L-DMEM was given in group H.The indexalso of right ventricle systolic pressure(RVSP),right ventricle hypertrophy index,arterial blood gas analysis and the changes of small pulmonary blood vessel were observed after 28 days.Results The administration of MMSCs 28 days after PAH nearly completely prevented the increase in RVSP with PAH alone [(32.20?2.32)mmHg vs(48.30?1.56)mmHg P

Eleven type 1 diabetic patients who received fixed regime of insulin Glargine were included in the study.The levels of fasting serum insulin were measured for each subject at 6:00 in three consecutive mornings.The variability of mean fasting serum insulin in each subject was 3.3%-41.5% (mean 15.4%).The variability did not correlate with the dose of Glargine statistically.

OBJECTIVE: To test droplet digital PCR for species identification and absolute quantification of biological sample. METHODS: Specific primers and probes for human mtDNA encoding gene ND4 and 16S rRNA were designed, and the species-specificity was assessed on DNA samples derived from human and common animals. To determine the sensitivity and stability of droplet digital PCR for species identification and absolute quantification, gradient dilution series of recombinant plasmid and 16 human DNA samples were analyzed. RESULTS: Human recombinant plasmid FAM (ND4) could be used in detecting the samples of human. And the results of detecting were consistent with all levels of diluted concentrations. Droplet digital PCR was able to detect low and single copy of target DNA. CONCLUSION Droplet digital PCR, with high sensitivity and specificity, is fully amenable for species identification and absolute quantification of biological samples, also it can be applied on routine forensic examination.
Animals ; DNA ; DNA Primers/genetics* ; DNA, Mitochondrial/genetics* ; Humans ; NADH Dehydrogenase/genetics* ; Polymerase Chain Reaction/methods* ; RNA, Ribosomal, 16S/genetics* ; Sensitivity and Specificity ; Species Specificity

OBJECTIVETo observe the histocompatibility of a polypropylene matrix implanted subcutaneously for potential hair follicle transplantation in rabbits.

METHODSThe polypropylene matrix for harboring the hair follicles was prepared and implanted subcutaneously at the neck of 5 New Zealand white rabbits by means of hair follicle unit transplantation. At 1 week after the transplantation and then on a monthly basis in the following 6 months, full-thickness skin tissues were sampled at the site of grafting to evaluate the histocompatibility of the matrix material using HE staining and scanning electron microscopy.

RESULTSAt 1 week after implantation of the matrix material, a small number of inflammatory cells and lymphocytic infiltration were observed around the graft, with mild hyperemia in the proliferative capillaries and mild inflammatory responses. In the following 6 months, the inflammatory cells and lymphocytes around the graft decreased obviously or even disappeared, and such graft rejection responses as tissue lysis and necrosis were not observed. A large quantity of collagen fibers were found to encapsulate the polypropylene material.

CONCLUSIONPolypropylene matrix graft has good histocompatibility with the rabbit subcutaneous tissue without producing obvious graft rejection responses, suggesting its feasibility for further experiments of hair follicle transplantation.

Twelve cases of Bartter's syndrome were reported and reviewed retrospectively.Usually vomiting was the first sympton in children,while fatigue was common in adults.Bartter's syndrome was characteristic of hypokalemia,metabolic alkalosis,elevations of plasma renin activity,serum angiotersinⅡand aldosterone and juxtaglomerular apperatus hyperplasia.Supplementation of potassium choloride was the main manner of therapy.

Nosocomial infection (NI) is one of the most significant complications arising after open heart surgery,and leads to increased mortality,hospitalization time and health resource allocation.This study investigated the morbidity,mortality,and independent risk factors associated with NI following open heart surgery.We retrospectively surveyed the records of 1606 consecutive cardiovascular surgical patients to identify those that developed NI.The NI selection criteria were based on the Centers for Disease Control and Prevention (CDC) guidelines.The term NI encompasses surgical site infection (SSI),central venous catheter-related infection (CVCRI),urinary tract infection (UTI),respiratory tract infection and pneumonia (RTIP),as well as other types of infections.Of 1606 cardiovascular surgery patients,125 developed NI (7.8％,125/1606).The rates of NI following surgery for congenital malformation,valve replacement,and coronary artery bypass graft were 2.6％ (15/587),5.5％ (26/473) and 13.6％ (32/236),respectively.The NI rate following surgical repair of aortic aneurysm or dissection was 16.8％ (52/310).Increased risk of NI was detected for patients with a prior preoperative stay ≥3 days (OR=2.11,95％ CI=1.39-3.20),diabetes (OR=2.00,95％=CI 1.26-3.20),length of surgery ≥6 h (OR=2.26,95％ CI=1.47-3.47),or postoperative cerebrovascular accident (OR=4.08,95％ CI=1.79-9.29).Greater attention should be paid toward compliance with ventilator and catheter regulations in order to decrease NI morbidity and mortality following cardiovascular procedures.

Adolescent ; Anesthetics ; therapeutic use ; Humans ; Male ; Polychondritis, Relapsing ; drug therapy

The auricular (including auricular acupoints) adjuvant diagnostic method, besides inspection (including dyeing method), palpation (including thermometric method), tenderness method (including impressing method) and electrical detection, includes the adjuvant diagnostic method of stimulating auricular points as well, it has been mostly studied and used by specialists of western medicine or doctors of integrated Chinese and western medicine. But it hasn't been introduced in the published Chinese monographs of auricular acupuncture yet. This article briefly introduces the adjuvant diagnostic method of stimulating auricular points combined with X-ray radiography; application in fetal heart electronic monitoring and fetal biophysical monitoring; and diagnostic methods of auricle reflex, vascular autonomous signals, and auricle and somatic 7 frequency response regions, which began to be researched abroad 35 years ago. The authors hope it will give some invigoration or illumination to my colleagues in acupuncture, especially those who are interested in auricular acupuncture.
Acupuncture Points ; Acupuncture, Ear ; Diagnostic Techniques and Procedures ; Fetal Monitoring ; Humans ; Medicine, Chinese Traditional ; Reflex

Autoimmune Diseases ; diagnosis ; etiology ; therapy ; Child, Preschool ; Encephalitis ; diagnosis ; etiology ; therapy ; Female ; Humans ; Receptors, N-Methyl-D-Aspartate ; immunology