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.

ObjectiveTo investigate the features of normal and psoriasis skin on ultrasound biomicroscopy (UBM) and explore the method of thickness measurement.MethodsUsing 50 MHz ultrasound probe of biological microscope, ultrasonographic observation and ultrasonic thickness measurement were conducted in 90 normal adults and 40 psoriasis patients. Innormal patients, ultrasound evaluations were performed at 10 different parts of the body skin.ResultsOn sonogram, the normal skin showed a “sandwich” structure with two parallel hyperechoic bands and the middle isoechoic dots and short liens. The sonograms of the psoriasis skin showed obviously thickened epidermis and dermis, disordered internal structure and clear boundary from adjacent normal skin. The range of the epidermis’ thickness measurement was between the medial forearm (0.12±0.03) mm and the palm (0.29±0.15) mm. The range of dermal thickness measurement was between the back hand (1.18±0.32) mm and parasternal (1.55±0.21) mm. Psoriasis skin was thicker than the uninvolved skin (P<0.001). And the dermis’ thickness of uninvolved skin in psoriasis patients was thicker than that of the normal adults (P<0.001).ConclusionNormal adult’s epidermis, dermis and skin appendages can be shown clearly using 50 MHz ultrasound biomicroscopy. And ultrasound biomicroscopy canaccurately measure the thickness of dermis and epidermis, which provides the basis for the diagnosis and treatment of psoriasis.

Objective To analyze the correlation between apolipoprotein E(ApoE) gene polymorphisms and the incidence rate of chronic cardiovascular disease as well as the blood lipid levels of patients.Methods ApoE gene polymorphism and lipid levels were measured by suing gene chip analysis system and biochemical analyze in 1 414 cases of chronic cardiovascular disease patients(experimental groups) and 374 cases of healthy subjects(control group).Results Compared with control group,E3/4 genotype frequency was increased in experimental group,while E2/E3 genotype frequency decreased(P<0.05).Compared with control group,the levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein-cholesterol(LDL-C) were obviously increased and the level of HDL-C was decreased in experimental group(P<0.05).Compared with patients with E2/E3 genotype,the level of HDL-C in patients with E3/E4 genotype was decreased and the levels of TC,LDL-C were increased significantly(P<0.05).Proportions of different ApoE genotypes in patients with cerebral infarction,cerebral hemorrhage,hypertension,coronary heart disease,type-2 diabete and fatty liver were different.Compared with the E2/E3 genotype,the proportion of the E3/E4 genotype in patients with cerebral infarction,cerebral hemorrhage,hypertension,coronary heart disease,type-2 diabete and fatty liver were increased(P<0.05).Conclusion ApoE gene polymorphism might be important cause of the individual difference of lipid levels and a risk factor for the occurrence and development of chronic cardiovascular diseases.

Objective To investigate the protective and therapeutic effect of chalcone ketones compound(code:L2H17,hereinafter referred to as L2)on mice infected with influenza A virus.Methods BALB/c mice were randomly divided into six groups:normal group,model group,positive drug-treated group,L2 treated groups (3 different concentrations).The mice were adapted for 72 hours,before a model was established by intranasal infection.Mice in each group were given medicine by i.g once daily for 6 days starting 24 h before virus challenge.Survival was observed daily for 14 days.The mortality,median survival time,rate of death protection and rate of prolonging life were determined to observe the therapeutic effect of chalcone(L2) against influenza virus infection.The whole lungs were taken under aseptic conditions on days 3 and 5 post-infection to calculat lung indexes and lung index inhibition.The left lung was fixed with 4% formaldehyde for pathological biopsy,the right lung was soaked in RNAstore to detect lung tissue viral load,and the double antibody sandwich ELISA method was used to detect the expression of inflammatory cytokines IL-6 and TNF-α in order to observe the therapeutic effect of L2 on viral pneumonia caused by influenza virus infection.Results Compared with the model group,the L2 80 mg/kg treatment group exhibited significant increases in median survival time(11 d),the rate of death protection (50%),and the rate of prolonging life(24.1%)but a moderate 50% decrease in mortality.In addition,the lung index decreased significantly both on d 3 and 5 after virus infection (P<0.05).The pathological results also improved significantly.The L2 80 mg/kg dose group had a significantly lower viral load of lung tissue on d 3 and 5 post-infection(P<0.05).Compared with model group,the expression of inflammatory factor IL-6 became lower to different degrees.Conclusion L2 has a protective effect on mice infected with influenza virus by reducing the degree of pathological changes of pneumonia caused by influenza viruses.