Objective To study the application value of T lymphocyte subsets and inflammatory factors in evaluating severity andprognosisofaplasticanemia.Methods 40patientswithaplasticanemiainthehospitalwereenrolledinthestudyasAAgroup, which was further divided into acute aplastic anemia(AAA) subgroup and chronic aplastic anemia(CAA) subgroup .In addition to that ,40 healthy people who received physical examination in the hospital during the same period were enrolled in the study as con‐trol group .Then blood routine tests were performed ,T lymphocyte subsets content in peripheral blood and serum inflammatory fac‐tor content were detected and the results were compared .Results Th1 ,Th17 contents and Th1/Th2 ,Th17/Treg ratio in AA group were higher than those in control group(P<0 .05) ,while Th2 ,Treg contents were lower(P<0 .05);Th1 ,Th17 contents and Th1/Th2 ,Th17/Treg ratio in AAA group were higher than those in CAA group(P<0 .05) ,while Th2 ,Treg cells contents were lower . Serum IL‐2 ,IFN‐γ ,IL‐17 contents in AA group were higher than those in control group(P<0 .05) ,while IL‐4 ,IL‐6 ,IL‐10 contents were lower(P<0 .05);serum IL‐2 ,IFN‐γ,IL‐17 contents in AAA group were higher than those of CAA group ,while IL‐4 ,IL‐6 , IL‐10 contents were lower .Hb ,PTL contents were negatively correlated with Th1 ,Th17 contents and serum IL‐2 ,IFN‐γ ,IL‐17 contents ,while positively correlated with Th2 ,Treg cells contents and serum IL‐4 ,IL‐6 ,IL‐10 contents .Conclusion Peripheral blood T lymphocyte subsets and serum inflammatory factors contents in aplastic anemia are abnormal and are correlated with the severity and prognosis related indicators .

Objective To investigate the time, location and relationship to clinical manifestation of abnormal expression of tyrosine hydroxylase(TH) in the cerebellum of Niemann\|Pick type C disease (NPC). Methods Immunohistochemical staining was applied, by using antibodies to TH, to brain sections from NPC and normal mice of various ages. Some adjacent sections were stained for calbindin D28k. Results There was no marked decrease in number of cerebellar Purkinje cells(PC), which were negatively stained for TH, in the NPC mice aged 1\|3 weeks. The PC were decreased in number starting from week 5. During postnatal weeks 8\|11, PC were significantly lost, but considerable number of PC in the nodulus and uvula vermis survived. Some survived PC and their dendritic trees were TH immunoreactive after week 8. These PC showed axonal spheroids and irregular dendrites that were bent, broken, locally enlarged or atrophied. Conclusion Mutation of NPC1 gene induces severe loss of cerebellar PC and survived PC have been damaged morphologically and show abnormal gene activity. These may be the pathogenic basis of movement disorders of NPC.

Objective To investigate the distribution of five indicators of hepatitis B(HBV-M)and its relationship with liver function parameters and HBV-DNA load in patients with chronic HBV infection in different stages.Methods The serum samples were collected from 456 patients infected with HBV.The HBV-M,liver functional parameters and HBV-DNA level were quantita-tively detected.According to the stages of disease,the patients were divided into 3 groups including chronic hepatitis B group(inclu-ding mild subgroup,moderate subgroup and severe subgroup),liver cirrhosis group(including compensatory subgroup,decompen-sated subgroup)and hepatocellular carcinoma goup.Results The ratio of HBsAg,HBeAg,HBcAb positive pattern(135 pattern) and HBsAg,HBeAb,HBcAb positive pattern(145 pattern)in the three groups were statistically different(P <0.05).In each chron-ic hepatitis B subgroup,both ALT and AST levels of 135 pattern were significantly higher than the other two patterns(P <0.05). In each liver cirrhosis subgroup and hepatocellular carcinoma group,there were statistically significant differences in ALB and TBIL levels between the three patterns(P <0.05 ).In each group,the HBV-DNA level of 135 positive pattern were significantly higher than the other two patterns(P <0.05).Conclusion With the advancement of chronic hepatitis B,there is a downtrend in the ratio of 135 pattern and increasing trend in the ratio of 145 pattern.If the stage of hepatitis B is discriminated,ALT,AST,ALB TBIL and HBV-DNA level were closely related to HBV-M pattern.

Objective To observe the variation of the blood pressure in patients with persistent vegetative state (PVS) during whirl-pool-bubble bath. Methods 5 PVS patients accepted whirlpool-bubble both therapy, 38℃for 20 minutes. Their blood pressure was recorded 0, 10 and 20 minutes after immersing for 4 weeks. Results The systolic pressure decreased 20 minutes after immersing compared with that just immersing, but all above 60 mmHg. The systolic and diastolic pressure maintained in a satisfactory range during whirlpool-bubble bath. Conclusion Hypotension has not been found in the patients with PVS during whirlpool-bubble bath.

The purified 3C8 was obtained by two step column purification,including Protein G affinity purification and DEAE anion exchange purification.The purity of purified 3C8 was about 93％ when analyzed by reverse column.SDS-PAGE showed that the purity of 3C8 was increased greatly by two step purification.By flowcytometry we found that 3C8 specifically binded with B7-H4/293T cells and did not bind with Mock/293T cells,moreover 3C8 did not bind with other B7 family members transgene cells.In confocal experiment 3C8 could specifically stained B7-H4/293T cells.In Western blot only B7-H4/293T cells showed positive band while Mock/293T cells showed negative result.The result of immunohistochemistry showed that B7-H4 was highly expressed in prostate cancer and renal cell carcinoma,while para-cancer tissues did not express B7-H4.The T cell proliferation experiment showed that B7-H4-Ig could bind to activate T cells and inhibit T cell proliferation,while 3C8 could block the binding of B7-H4-Ig and reverse the T cell proliferation inhibition effect of B7-H4-Ig by CFSE and CCK8 assay.The cytokine IFN-γ and IL-2 secreted by activating T cells was decreased by B7-H4-Ig and 3C8 could reverse the effect of B7-H4-Ig.

A series of phthalazine ketone compounds were synthesized and the structures were confirmed by H NMR and HR-MS spectrum. All target compounds were obtained through 7 steps, including selective reduction, nitration, bromination, ring enlargement, reduction, Knoevenagel and acylated reaction. The compounds were evaluated for their immunosuppressive effects of T-cell proliferation and inhibitory activity of IMPDH type II in vitro, as well as their structure-activity relationship were assessed. Several compounds exhibited strong immunosuppressive properties, especially compounds 7f and 7h, with IC50 values of 0.093 micromol x L(-1) and 0.14 micromol x L(-1) respectively, which were superior to mycophenolic acid. The information obtained from the studies may be useful for further research on the immunosuppressive agents.

Objective:To develop a two-step method for purification of monoclonal antibody rhTF243 protein from mouse ascites by using hydrophobic charge induction chromatography(HCIC) and affinity chromatography with protein A sepharose CL-4B.Methods:The ascites was first purified by HCIC after centrifugation and filtration.Then the fraction containing the protein of interest was directly purified by affinity chromatography with protein A sepharose CL-4B.Results:The purity of the obtained monoclonal antibody was up to 97% with recovery of 73% and of high activity.Conclusion:The method for purification of monoclonal antibody is developed using HCIC and Protein A affinity chromatography and the obtained antibodies are of high purity and activity.

OBJECTIVE To investigate whether the KCNN4 gene and KPTN gene contribute to a Chinese non-syndromic hearing loss pedigree linked to DFNA4 with positional candidate approach. METHODS The complete coding region of the two genes were amplified with polymerase chain reaction (PCR), and bidirectional sequencing of the PCR products was subsequently applied in the 36 family members to identify the possible mutations or polymorphisms in the candidate genes. RESULTS Sequence analysis of coding regions and splice sites of the two candidate genes in 36 members including 12 hearing-impaired individuals in family Z002 failed to demonstrate any deafness-causing mutations of KCNN4 gene. There was one heterozygous mutation identified in exon10 coding sequence (942C/T) of KPTN gene, which did not result in amino acid change (P302P) as a repoerted synonymous SNP site (rs2293424). This SNP site did not cosegregate with the phenotype of family Z002. CONCLUSION Our study excluded the two candidates, KCNN4 and KPTN , as the causative genes involved in this Chinese DFNA4 pedigree.

AIM: To study the expression of connective tissue growth factor (CTGF) induced by hypoxia, and the role and mechanism of hypoxia on promoting renal interstitial fibrosis. METHODS: Renal interstitial fibrosis was induced by unilateral ureteral obstruction (UUO) in rat animal model for 9 days as in vivo studies; marker of hypoxia-HIF-1? mRNA and protein, the expression of CTGF in the obstructed kidneys were assessed by RT-PCR, immunohistochemistry and Western blotting respectively. In vitro , normal rat kidney interstitial fibroblast cells (NRK-49F) were exposed to hypoxia (1%O 2) for up to 6 hours, hypoxia was confirmed by detecting the expression of HIF-1? protein in cells, cellular level of CTGF mRNA and protein were assessed by RT-PCR and Western blotting respectively. RESULTS: Neither HIF-1? mRNA nor HIF-1? protein was expressed in the kidney from sham-operated group of rats. High level of HIF-1? mRNA were occurred, and strongly HIF-1? positive immunostaining were seen in the tubular and interstitial cells in kidney from UUO rats. Expression and location of CTGF protein were paralleled and relevant with the expression of HIF-1? protein in kidney of UUO rats. In cultured NRK-49F cell line, subjected to hypoxia even for 6 hours stimulated the expression of CTGF mRNA and protein. CONCLUSION: Our results indicated that hypoxia could stimulate the expression of CTGF mRNA and protein in kidney from UUO rats, which may in turn contribute to renal interstitial fibrosis.

Objective:To assess the possibility of suicide in setting condition among urban and rural residents samples in northern China.Methods:By randomly sampling,1007 persons aged 18 years or older were selected from 10 urban neighborhoods in Beijing and from 10 rural village in Hebei province.The Survey Schedule of Atti-tudes in China in which 24 negative life events were listed,and the possibility of committing suicide was asked to the subjects when they encountered the supposed events.Results:The rates of thinking about committing suicide when encountered the supposed life events were 65.2% of incurable illness,62.0% of burden on other and no future hope,61.2% of drug dependence,57.3% of large debt due to gambling,54.2% of being raped, 51.6% of severe depression,50.7% of being elderly and no family to provide support.Only 14.6% of the ur-ban and rural residents would not consider suicide when encountered any type of supposed problems.Conclusion:There is higher possibility of considering suicide in urban and rural residents when encountered negative life events, so it is important to give support to the residents with one or more life events.