Humans ; Traumatology

In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.

To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.
Animals ; Antibodies, Viral ; blood ; Chikungunya Fever ; blood ; diagnosis ; virology ; Chikungunya virus ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; blood ; Mice

Animals ; Bone Marrow Cells ; cytology ; Brain ; cytology ; physiopathology ; Brain Injuries ; physiopathology ; therapy ; Cell Movement ; Cells, Cultured ; Cisterna Magna ; Immunohistochemistry ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; transplantation

Objective To assess the effective length of jejunal graft when the 3~(rd) intestinal artery is u- tilized as vascular pedicle and afford a reliable theoretic base for clinical esophageal reconstruction.Methods In 32 formalin preserved and 21 fresh cadaver specimens,the diameter of 1st to 5th intestinal arteries and diameter of arterial arches are measured with linear calibre.Measure the length of jejunum that can be harves- ted as graft when the arches are extended.In the 21 fresh specimens,the 1st,2nd,4th and 5th intestinal ar- teries are ligated,acetic ester stained with red dye were injected into the lumen of 3rd intestinal artery via catheter.Extent of distribution of the arteries to the jejunum was observed.And then red ABS solution was in- jected into the 3rd intestinal artery to make into cast specimen.The blood supply distribution of jejunum through 3rd intestinal artery-arterial arch and communicating system were observed again.Results The di- ameter of the 3rd intestinal artery was the largest among the 1st to 5th intestinal arteries.The length of jejunum vascularized by 3rd intestinal artery can be as long as (142.2?62.3) (69.0～206.60cm) in acetic ester in- filtrated specimens.While in ABS east specimen,the average available extent of donor jejunum was(30.8?7.3) (23.0～37.3cm).Conclusion As observed by this applied anatomy study,the jejunum graft vascu- larized by 3rd intestinal artery alone has sufficient length to meet the need of esophageal reeonstrution.

Objective To investigate the significance of the serum levels of interleukin-10 (IL-10),IL-13,IL-15 of patients with chronic hepatic failure and the correlation between those inter- leukin levels and nosocomial infections.Methods The serum levels of IL-10,IL-13,IL-15 of 58 patients with chronic hepatic failure were measured by double antibody sandwich enzyme-linked immu- nosorbent assay at the time of admission and 2 weeks after admission.Results The serum levels of IL-15 and the propotion of IL-15/IL-10 and IL-15/IL-13 in patients with chronic hepatic failure group at the time of admission were significantly higher than those in healthy control group[(358.16?290.91) ng/L vs (38.55?21.49) ng/L,12.93?14.26 vs 1.10?0.55,98.55?97.5.5 vs 9.70?5.03,respectively,all P=0.000].Those in death group were significantly higher than those in improving group[(479.93v205.52) ng/L vs (244.51?236.29) ng/L,17.65?17.78 vs 8.53?7.98,130.69?115.50 vs 68.55?65.99,respectively,all P

0.05).However,the shorter the history of the disease,the better the efficacy of the treatment.The younger the patient was,the better the efficacy of the treatment.Conclusions Vulva dystrophy can be treated with focused ultrasound effectively and safely.This approach appears to be a new promising treatment method.

OBJECTIVETo study the effect of 5-fluorouracil-FU in combination with astragalus membranaceus(AM) on amino acid metabolism in mice model of gastric carcinoma induced by 3-methylcholanthrene(MC).

METHODSMice gastric carcinoma models were established by 3-methylcholanthrene induction and randomly divided into different groups, and received 5-FU treatment (group A) 5-FU plus AM (group B), 5-FU plus a high dose of AM(group C), no treatment (group D). Normal mice were used as control (group N). Free amino acid in the tumor specimens were examined.

RESULTSThe levels of free Valine, Methionine, Leucine, Arginine and cystine in the tumor specimens in group D were significantly higher than that in group N(P< 0.05). The levels of free serine in group A, B, C, D were significantly higher than that in group N. The levels of free glutamic acid in group A, B were significantly higher than that in group N(P< 0.05). The levels of free proline in group C, D were significantly higher than that in group P, N(P< 0.05).

CONCLUSIONSThe increasing levels of free serine and proline in tumor specimens in gastric cancer mice model reveals metabolic disturbance of amino acid. 5-FU plus astragalus membranaceus can decrease the level of free glutamic acid in the mice models, and inhibit tumor growth.

Amino Acids ; metabolism ; Animals ; Astragalus membranaceus ; Fluorouracil ; therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Phytotherapy ; Stomach Neoplasms ; drug therapy ; metabolism

Objective: To establish a method for isolation and purification of fetal membrane derived adherent cells (FMDACs) , and investigate their biological characteristics. Method: FMDACs were isolated with trypsin inducing and cultured in vitro. FMDACs were induced to differentiate into osteoblasts and adipocytes. FACS and immunocytochemistry technique were used to examine the cell surface antigen. The genetic stability was verified by karyotype analysis. Results: FMDACs were successfully isolated and expanded in vitro. They had strong proliferative ability. FMDACs were positive for CD44 and CD29, but negative for CD34, CD14 and CD45. FMDACs were differentiated into osteoblasts and adipocytes after inducement. The karyotype was stable in the sixth-passaged FMDACs and the tumorigenicity was not found. Conclusion; FMDACs have the possibility of multipotent stem cells, which have strong capacities of self-renewal and multidirectional differentiation. The genetic background of FMDACs is stable. FMDACs may be used as a kind of novel seed cells for tissue engineering.

Objective To investigate the effect of a selective muscarinic receptor antagonist (penehyclidine hydrochloride) in three vessel occlusion model of acute global cerebral ischemia-reperfusion in rats.Method One hundred and forty-four male SD rats were randomly divided into four groups:sham operated group,vehicle treated group (saline 1 ml,i.p.),scopolamine treated group (0.01 mg/kg,i.p.) and penehyclidine hydrochloride treated group (0.01 mg/kg,i.p.) with drugs injected 40 minutes before ischemia respectively.The ischemic duration was 10 minutes.The animals were subjected to motor activity tests (open field activity test,beam-walking test and grip test) at 24 hours or on the 3rd and 7th day after reperfusion.HE staining,TUNEL staining and immunohistochemical reactions of bax and bel-2 were carried out at the time points of 2,12,24 hours,3 and 7 days after reperfusion.TTC staining was carried out in some rats for assessment of infarction volume on the 4th day after reperfusion.Results As compared with the vehicle treated group,both penehyclidine hydrochloride treatment and scopolamine treatment decreased the numbers of apeptotie neurons (P