A batch affinily adsorption method with heparin-sepharose 4B gel has been described for preparation and purification of antithrombin Ⅲ from plasma. The me- thod is suitable for batch preparation. The processes of each batch are completed only in 3 days. The recovery of activity was 22.4-42.3% from 20 liters of plasma. By fur- ther purification, the recovery of activity and antigen was 26.3% and 20.3% respec- tively, the specific activity was 9.9U/mg. The purified AT-Ⅲ showed a single band on disc gel electrophoresis, and the levels of purity, biologic activity and the recov- ery of activity are similar to those of for- eign reports.

Aim To investigate whether the annexation of bradykinin and papaverine had some synergetic effects on the opening of blood-tumor barrier,and find out the action of cytokine in this mechanism.Methods 160 female rats were divided into 8 groups:sham operated group;model group,PA group,BK group,PA+BK group,1/2PA+BK group,1/2BK+PA group and 1/2PA+1/2BK group.20 rats were needed in each group.The C6 brain tumor model was built.The drug was pumped into rat's brain via the carotid artery.The opening of the tight junction was detected by electron microscop.And the permeability of the blood-tumor barrier was tested by Evans blue.The expression of IL-1? was tested by Western blot and the expression of IL-1? on/around the capillary in the brain of the cerebral glioma rats was tested by SABC immunochemistry.Results In contrast to sham operated group/model group,the expression of IL-1? in PA group,BK group and PA+BK group increased obviously(P

BACKGROUND:Vaginal smooth muscle cells are mainly cultured by enzyme digestion method and explant method.The former method requires a short culture time with a high output,but this method demands a large number of tissues with a high opportunity of pollution,and the optimal digestion time is difficult to control.The latter method is simple and effective,but the culture needs a long time.OBJECTIVE:To observe the culture time of rabbit vaginal smooth muscle cells using explant + enzyme digestion methods,and to compare with the explant method.DESIGN,TIME AND SETTING:The in vitro observation experiment was performed at the Burn Institute and Animal Laboratory of First Affiliated Hospital,Nanchang University from February 2005 to February 2006.MATERIALS:Female New Zealand rabbits aged 4-5 months,with the body mass of 2.0-2.5 kg were selected for this study.METHODS:Vaginal smooth muscle cells using explant method:1 mm?1 mm?1 mm explants were uniformly incubated in a culture medium,and then placed in a incubator at 37 ℃ for 3.0-4.0 hours.After tissue confluence,tissues were immersed in DMEM containing 0.1 volume fraction,stored at 37 ℃ for 3.0-4.0 days.Following cells grew from the tissue islets,the medium was changed about once every three days.Vaginal smooth muscle cells using explant + enzyme digestion methods:The tissue pieces were digested by 0.1% collagenase type Ⅳ at 37 ℃ for 0.5-1 hour,and then enzymatic digestion was interrupted when the edge of tissue pieces became coarse.These pieces were cultivated in culture dishes.The remaining steps were the same as the explant method.Serial subculture:The 1st subculture was made when 50%-60% cells were confluent.After the 2nd passage of cells was obtained,subculture was made when 80% cells were confluent.MAIN OUTCOME MEASURES:Time of primary culture;Surface structure and ultrastructure of the 3rd passage of vaginal smooth muscle cells.RESULTS:The eruption time and the confluent time of vaginal smooth muscle cells could be shortened by the explant + collagen digestion methods about 1-2 days and 3-4 days respectively compared with the explant method only.Vaginal smooth muscle cells,which were obtained by the two methods mentioned above,could propagate for 5-6 passages.The attached cells were polygonal or spindle and after confluence could be seen,the marked smooth muscle cells "Peak-Valley" structure in some domains under the inverted microscope.The third passage of smooth muscle cell nuclei were serration;the cytoplasm contained the marked smooth muscle cell structural myofilaments and dense bodies and plenty of golgi bodies;the rough endoplasmic reticulum broadened;the free ribosome were abundant under the transmission electron microscope.This indicated that vaginal smooth muscle cells with synthesis phenotype could be greatly collected.CONCLUSION:Explant + enzyme digestion methods need a short cycle of primary culture.Vaginal explants should be kept moisture during drawing materials,isolation and the whole.The time of enzymatic digestion should not be too long,to the point that palpi pilosi is seen surrounding the explants.Primary culture should remain static state and use of plastic culture dishes.

Objective To interpret the clinicopathological features and the key factors for diagnosis of polyomavirus-associated nephropathy (PVAN).Methods Clinicopathological data of 13casesof polyomavirus-associatednephropathyduring2008-2011inour hospitalwere retrospectively analyzed.Three cases received repeat biopsy.The clinicopathological features were analyzed according to thelight microscopicsceneandSV40-Timmunochemicalexpression.Results Recipients had a peak incidence of PVAN in 12 to 18 months period after renal transplantation,accompanied by elevated serum creatinine.Due to the progression of the disease,3patterns of histological findings could be identified.The early lesion was confined to the collected ducts,with slightly inflammatory infiltration in medullary interstitium,viral inclusions were not necessarily seen.The only findings could be enlarged nuclear and irregular arrangement of the tubular epithelial cells.At the developing stage,prominent tubulointerstitial nephritis was detected,and the involved tubules extended to other segments of renal tubule,even the parietal epithelial cells of Bowman's capsule could be compromised.The epithelial cells shed off,leading the tubular basement membrane exposed.Typical intra-nuclear inclusions as well as variable nuclear changes were found.At the end stage,the allograft showed notable chronic tubulointersititial change,with diffuse tubular atrophy and interstitial fibrosis.Although in this period,typical viral inclusions were rare, stillIHCshowedpositiveexpression of SV40-T. After immunosuppressantreductionor exchange,2 cases developed renal failure,4 cases showed sustained increment in serum creatinine,while 7 cases had a stabilized serum creatinine level.Conclusions Polyomavirus-associated nephropathy can display uneven pathological changes,as well as the morphology of the infected epithelial cells.Segments of the involved tubule are associated with the course of disease.Reduction of immunosuppressant at the early stage has a favorable effect.A prompt renal biopsy should be done in renal transplant recipient if who shows increased serum creatinine,and a routine polyomavirus immunohistochemical staining should be applied as well.

Purpose:To investigate the relationship between the expression tansforming growth factor ?(TGF ?) and the differentiation, invasion of colorectal adenocarcinoma.Methods:To observe the expression of transforming growth factor ? and transforming growth factor ? receptor in 68 samples of colorectal adenocarcinoma with immunohistochemical technique. Results:The cells with positive reaction of TGF ? antibody were scattered in the stroma. The cells with infiltrative growth had positive TGF ? staining.Conclusions:It was indicated that TGF ? might play an important role in the invasion and metastasis of colorectal adenocarcinoma.

Objective To explore the establishment of peptide mapping database of Candida albicans ,laying the foundation for rapid diagnosis of Candida albicans infection .Methods 96 Candida albicans were collected clinically ,and its DNA was extracted . Polymerase chain reaction(PCR) was used to amplify the ITS1-5 .8S-ITS2 gene fragments and restriction endonucleases were a-dopted to identify them .Surface enhanced laser desorption ionization-time of flight-mass spectrometry(SELDI-TOF-MS) instrument was applied to detect the Candida albicans peptide mapping ,and Ciphergen ProteinChip software was used to collect data automati-cally .The established peptide mapping database was verified by confirmed Candida .Results According to restriction fragment length polymorphism analysis ,96 strains were confirmed as Candida albicans .15 peptide peaks were captured by SELDI-TOF-MS chips .Five peptide peaks of them with stable expression were screened out ,and the similarity analysis software was used to estab-lish peptide mapping database of Candida albicans .More than 95% of similarity was found between peptide mapping of Candida albicans and established database ,while less than 50% was found between peptide mapping of other Candida species and database . Conclusion The establishment of peptide mapping database of Candida albicans provides a theoretical basis for the rapid diagnosis of Candida albicans infection .

OBJECTIVE To understand the pathogens and their drugs resistance in general surgery department and provide rational suggestion of antibiotics use for clinic treatment.METHODS A total of 158 cases with nosocomial infection among the general surgery department inpatients from Jun 2006 to Oct 2008 were retrospectively analyzed and summarized.RESULTS The common nosocomial infection sites were the lower respiratory tract,gastrointestinal tract,urinary tract and surgical sites.The G-bacilli of nosocomial infections in turn were Escherichia coli(18.02%),Acinetobacter baumannii(11.26%),Pseudomonas aeruginosa(7.66%),and Klebsiella pneumoniae(4.50%).The main G+cocci were Staphylococcus aureus(21.62%),Enterococcus faecium(5.86%)and E.faecalis(3.15%)in turn.In G-bacilli,the sensitivity to imipenem was the highest from 58.82% to 100.00%.The sensitivity to amikacin were more than 70.00% except A.baumannii,and to sulbactam/cefoperazone were more than 50.00% except Pseudomonas aeruginosa.In G+ cocci,the sensitivity to vancomycin of S.aureus and E.faecium was 100.00% and 84.62%.CONCLUSIONS Investigating the pathogens and their drug resistance in general surgery department is very important to prevent and control nosocomial infections.

B ecause of the exist of com plex m atrix, the confirm ing indicators of qualitative results for toxic substances in biological sam ples by chrom atography-m ass spectrom etry are different from that in non-biological sam ples. E ven in biological sam ples, the confirm ing indicators are different in various ap-plication areas. T his paper review s the sim ilarities and differences of confirm ing indicators for the ana-lyte in biological sam ples by chrom atography-m ass spectrom etry in the field of forensic toxicological analysis and other application areas. T hese confirm ing indicators include retention tim e (R T ), relative re-tention tim e (R R T ), signal to noise (S/N ), characteristic ions, relative abundance of characteristic ions, parent ion-daughter ion pair and abundance ratio of ion pair, etc.

Objective To explore the role of cell proliferation and apoptosis, and the expression significance of involved protein as Bcl-2(B-cell lymphoma/leukemia-2) and Bax (Bcl-associate X protein) in the developing small intestine of human fetus. Methods The expression product of Bcl-2 and Bax was investigated with immunohistochemical methods in the 2nd, 3rd and 4th months of gestation respectively. Results In the second, the third and the fourth month of gestation, the Bcl-2 immunoreactive positive signals were found in the ganglion cells of intermuscular and submucous nerve plexus. Bax positive cells were observed in the cytoplasm of the simple columnar epithelium cells of the small intestinal mucous layer.Conclusion Bcl-2 and Bax proteins regulate the developing small intestine of human fetus.

Objective To study the clinical characteristics and the influencing factors of hypertension in IgA nephropathy(IgAN)patients with normal renal function.Methods From 2002 to 2006,a total of 507 idiopathic IgAN patients with normal renal function(eGFR≥90 mL/min)confirmed by renal biopsy were treated in the First Affiliated Hospital of Sun Yat-sen University.The patients were divided into a hypertension group(n=93)and a normal group(n=414)according to the BP levels.Univariate and multivariate logistic regression analysis were used to analyze the relationship between the hypertension and the clinical characteristics.Results We found that 18.3%(93/507)of the IgAN patients had hypertension,with hypertension as the main symptom in some cases.Univariate analysis showed that male sex,older age,higher BMI,elevated level of triglyceride and cholesterol were the clinical risk factors for hypertension in IgAN patients(P