Phospholipid fatty acids are the major constituents of the membranes of all living cells, and different groups of microorganism synthesize a variety of PLFA through various biochemical pathways. Several PLFAs can be used as "signatures" to analyze changes in microbial biomass and microbial communities structure. More and more PLFA method was used in soil microbial analysis. This article briefly reviewed the applications of PLFA methods in soil microbial analysis.

The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72～84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.

· AIM :To investigate the role of pericytes in growth of retinal microvascular endothelial cells with a co-culture system in order to understand some mechanism of angiogenesis in hypoxia induced retinal neovascular disorders.(RMECs) were isolated by a modified protocol using CD31 coated Dynabeads, and identified by immunocytochemical staining with anti-Factor Ⅷ and CD31 antibodies. Rat retinal pericytes were isolated and characterized by immunofluorescent staining with PDGFR-β; and desmin antibodies. Pericytes and RMECs were cultured in a contact co-culture system both under normoxia and hypoxia by Millicell chamber. RMECs proliferation was evaluated by MTT and cell cycle assay with flow cytometry. RT-PCR was used to detect the alteration of KDR/Flk-1 mRNA level in RMECs under normoxia or hypoxia in the co-culture system.harvested with the modified isolating method. The two cell types were identified by positive Factor Ⅷ, CD31 and PDGFR-β, desmin cytochemical staining respectively.RMECs proliferated significantly under hypoxia from 3 to 9d with a maximal rate on day 6 (24.9%, P ＜ 0.01) by MTT. In the co-culture system, the proliferation of RMECs was inhibited by pericytes. After 6d exposure to hypoxia,the fraction of S-phase RMECs number was greatly increased by 43.9% (P ＜ 0.01). In the co-culture system,RMECs proliferation was inhibited by pericytes through decreasing the fraction of S-phase cell number both under normoxia (3.6%, P＜0.05) and under hypoxia (15.1%,P＜0.01). KDR/Flk-1 mRNA level in single cultured RMECs was shown to increase approximately 1.3-fold when exposed to hypoxia. Compared with single cultured RMECs, co-culture with pericytes could decrease KDR/Flk-1 mRNA by 45.1% (P＜0.05) and 27.7% (P ＜ 0.05) under normoxia and hypoxia condition respectively.pericytes could inhibit proliferation of RMECs under both normoxia and hypoxia. The inhibition effects of pericytes maybe, at least in part, due to downregulation of KDR/Flk-1 of RMECs. These findings confirm that pericytes could be a potential inhibitor in the pathogenesis of retinal neovascularization.

Objective:To explore CCR7 expression in splenic dendritic cells and its role in migration and activity of splenic dendritic cells in multiple organ dysfunction syndrome (MODS) in mice.Methods:The MODS model of mice was reproduced by Zymosan injection into peritoneal cavity.The mice were randomly divided into groups of normal,3-6 hours,24-48 hours and 10-12 days post zymosan injection.CD11c and CD205 were analysed by immunohistochemistry;The expression of CD86 and CCR7 of DCs were studied by the flow cytometry analysis.Results:In normal mice,many DC were found at the margin between the red and white pulp.In the 3-6 h and 24-48 h groups,CD86 and CCR7 were strongly up-regulated in the DC,and they distributed in T cells areas.In the 10-12 d group,DC distributed at the margin by the immature form.Conclusion:The CCR7 expression level of DC has close correlations with the migration of DC,CCR7 can be used to evaluate the migration and functional activity of DC in MODS.

Abortion, Missed ; chemically induced ; Adult ; Carbon Disulfide ; adverse effects ; Female ; Humans ; Occupational Exposure ; adverse effects ; Pregnancy ; Retrospective Studies ; Risk Factors ; Time Factors

To study the tissue distribution and excretion of indomethacin 5-fluorouracil-1-ylmethyl ester (IFM) metabolite 5-fluorouracil in rats, an accurate and specific high performance liquid chromatography method for quantifying IFM in rat plasma and tissues was developed. Biological samples were prepared by liquid-liquid extraction and separated on a Diamonsil C18 column (250 mm x 4.6 mm ID, 5 microm). The mobile phase for tissue samples, plasma samples and feces samples were composed of methanol-water-36% acetic acid (3:96.9:0.1, v/v) and the mobile phase for urine samples was a mixture of methanol-water-36% acetic acid (10:89.9:0.1, v/v). The eluate was monitored by UV absorbance at 260 nm. After a single ig dose of 100 mg x kg(-1) IFM in rats, 5-Fu was mainly distributed in stomach, small intestine, and liver. The concentrations of 5-fluorouracil in other tissues and plasma were low. The excretion of 5-Fu in urine and feces amounted to 0.0065% and 0.063% of the dose, respectively. The method is shown to be accurate and specific, and suitable for preclinical pharmacokinetic studies of IFM.
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; metabolism ; pharmacokinetics ; urine ; Antimetabolites, Antineoplastic ; pharmacokinetics ; urine ; Feces ; chemistry ; Female ; Fluorouracil ; pharmacokinetics ; urine ; Indomethacin ; metabolism ; pharmacokinetics ; urine ; Male ; Prodrugs ; pharmacokinetics ; Random Allocation ; Rats ; Rats, Wistar ; Sensitivity and Specificity ; Tissue Distribution

Objective To report the outcome of emergency repair of severe complex defect in forearm by transplantation of free flap and simultaneous functional reconstruction.Methods From Mar.1994 to Aug.2003,4 cases with severe complex defect in forearm was repaired by transplantation of free skin flap, free skin flap combined with fibula flap,or fibula osteocutaneous flap in emergency.Simultaneously the flexion and extension function were repaired by muscle transfer and/or tendon grafting,tenonectomy.Results All the cases were successful.Follow-up period ranged from 1 to 3 years postoperatively.The blood-supply,tex- ture and elasticity of transferred flaps were excellent with good bone healing.Opposition of thumb with four fin- gers was good.Sensory recovery of the hand was satisfactory.Conclusion Transplantation of free flap com- bined with simultaneous functional reconstruction is an ideal method in emergency repair of severe complex de- fect in forearm.

Objective To evaluate the association of islet autoantibodies [ glutamic acid decarboxylase antibody(GADA),protein tyrosine phosphatase antibody(IA-2A)and insulin autoantibodies(IAA)1 with HLA- DQ genotypes in the first-degree relatives of autoimmune type 1 diabetes mellitus.Methods This was a cross- sectional and case-control study.Three hundred and fifty-one first-degree relatives with normal glucose tolerance of patients with type 1 diabetes mellitus and 376 healthy controls were recruited and measured for GADA,IA-2A and IAA by radioligand assay,and 156 first-degree relatives of patients with autoimmune type 1 diabetes mellitus and 278 controls were typed for genetic polymorphisms of HLA-DQ with PCR sequencing-based typing method.Results (1)DQA1*03,DQBI*0303,*0401 alleles and DQA1 * 03-DQBI * 0303,DQA1 * 05-DQBI * 0201,DQA1 * 03-DQBI * 0401 haplotypes were significantly increased in the first-degree relatives of autoimmune type 1 diabetes mellitus(P

Objective To assess the continent diversion results of sigma rectum pouch after radical cystectomy. Methods The reconstruction of bladder with sigmoid was modified for treatment of 18 cases of bladder tumor.The intestine was incised over a length of 20～24 cm with the junction of sigmoid colon and rectum as the midpoint so as to create a low pressure reservoir for urine and side-to-side anastomosis was performed on the posterior borders of the rectosigmoid wall.Submucosal tunnel modified technique was em- ployed in antireflux urethral implantation,Urination has been controlled by anal sphincter.Results About 80 minutes was spent to finish a new low pressure pouch after radical cystectomy.Among 18 patients with this op- eration,the controlled emiction were good after pull out the anal duct and"J"stent in 1 week to 2 months.Af- ter 2 months,the times of urination is stable,4～5 times in daytime and 1～3 times during nighttime.Two pa- tients had nocturnal enuresis and the symptom vanished after 2 months. One patient had adhesive ileus, two patients had hyperchloremia acidosis and kaliopenia,one patient had urethral stump cancer.There is no com- plication as anastomotic block,renal function lesion and severe upper urinary tract infection. Conclusion This operative method was easy,emiction control was well,and with higher quality of life for patients.It is al- so a better alternative diversion procedure that would be easily accepted.

OBJECTIVEThe goal of this study was to investigate whether murine cytomegalovirus (MCMV) is able to exacerbate the atherosclerotic process in apolipoprotein E knockout (apoE -/-) mice, and the effect of fluvastatin on the atherogenesis.

METHODSThe apoE-/- mice kept on a west diet were given low dosage of MCMV. At 14,18 and 24 weeks post infection, AS lesion were measured on aorta. The fluvastatin was administered, and AS lesion were measured accordingly above.

RESULTSWe observed that in the chronic phase of the infection, AS lesion area was significantly increased. MCMV gB mRNA was not amplified by real-time PCR from the arterial wall. The IgG antibody level of MCMV in blood plasma and the content of virus DNA in salivary gland were not correlated with AS lesions. After the administration of fluvastatin, there was no significant difference of AS lesions between MCMV infected group and mock-infected group.

CONCLUSIONMCMV may aggravate the AS lesion in apoE -/- mice in the chronic phase of infection, and promote more severe type of AS lesions. But it might not be the direct effects of mechanism of MCMV on the local lesion of AS. Fluvastatin could meliorate the progression of AS after MCMV infection, but this was not accomplished by decreasing MCMV duplication.

Animals ; Aorta ; drug effects ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; blood ; drug therapy ; genetics ; virology ; Fatty Acids, Monounsaturated ; pharmacology ; Herpesviridae Infections ; blood ; drug therapy ; virology ; Immunoglobulin G ; blood ; Indoles ; pharmacology ; Male ; Mice ; Mice, Knockout ; Muromegalovirus ; genetics