Tryptophan is the essential amino acid in the growth of cell.Indoleamine 2,3-dioxygenase is the rate-limiting enzyme in the catabolism of tryptophan along the kynurenine pathway.Indoleamine 2,3-dioxygenase enzyme mainly promotes tryptophan consumption of local micro-environment,causing tryptophan lack,thus inhibiting proliferation and activation of T cell.Tryptophan-derived catabolites suppress allogeneic T-cell proliferation (kynurenine,3-hydroxykynurenine,and 3-hydroxyanthranilate).Indoleamine 2,3-dioxygenase dendritic cells may interact with regulatory T cells forming an immunomodulatory network to promote immune tolerance induction.Though the immune regulation mechanism of the indoleamine 2,3-dioxygenase-initiated L-tryptophan in transplantation remains unclear,Numerous researches indicate that its role of tolerance induction in transplantation will be significant.

Objective To explore the expression of brain-derived neurotrophic factor(BDNF) and highaffinity receptors tropomyosin receptor kinase B(TrkB) protien in the prefrontal cortex of the post stroke depression in the rats.Methods Focal cerebral ischemic rat models were made with thread embolization method.Post stroke depression rat models were established with comprehensive separately breeding and chronic unpredicted mild stress (CUMS) on this basis.Normal control group,depression group and stroke group were used to compared with PSD group.6 rats were used in each group.Immunohistochemistry for detecting the expression of BDNF and TrkB in the prefrontal was used at 29th day since the CUMS.Results The number of BNDF immunopositive cells in PSD group was the least ((21.00 ± 12.41) per microscope field of vision) than other groups.Whereas there was no statistical difference among groups(P＞ 0.05).The number of TrkB immunopositive cells in the prefrontal cortex in PSD group was the least (20.78 ± 7.20) among three groups,and depreesion group was secondary (21.00 ±5.61).Stroke group has the most number of immunopositive cells(31.67 ± 7.38) in the prefrontal cortex among four groups.One way ANOVA statistical analysis showed the number of TrkB immunopositive cells decreased significantly in the PSD group and depression group compared with stroke group (P＜0.05).Conclusion The downregulation of TrkB immunopositive cells in the prefrontal cortex may be responsible for the pathogenesis of PSD.

Objective To explore the expression of brain-derived neurotrophic factor(BDNF) mRNA and high-affinity receptor TrkB mRNA in the prefrontal cortex of the post stroke depression in the rats.Methods Focal cerebral ischemic rat models were made with thread embolism method.Post stroke depression(PSD) rat models were established with comprehensive separately breeding and chronic unpredicted mild stress (CUMS) on this basis.Normal control group,depression group and stroke group were used to compare with PSD group.8 rats were used in each group.RT-PCR was employed to detect gene expression of BDNF and TrkB.GADPH was used as control at 29th day after the CUMS.Results The results showed that the gene level of BNDF in the prefrontal cortex of rat subjected PSD was lowest among all groups(0.75 ± 0.21).And the expression of BNDF mRNA in the normal control rats was (0.83 ± 0.16) and was highest among all groups.While it was (0.77 ± 0.22) in the depression group and (0.80 ± 0.20) in the stroke group.The one-way analysis of variance showed the expression of BDNF mRNA in the prefrontal cortex decreased significantly in the PSD group compared with normal control rats (P ＜ 0.05).Whereas,the expression of TrkB mRNA had no statistical difference among groups (P ＞ 0.05).Conclusion The downregulation of BDNF mRNA in the prefrontal cortex may be responsible for the pathogenesis of PSD.

[Objective]To compare the osteoinductive activity of calcium sulfate(CS),allogenetic demineralized bone materials(ADBM) and heterogenetic demineralized bone materials(HDBM) by observing their efficiency in inducing bone formation.[Method]CS,ADBM and HDBM were transplanted into thigh muscle pouches of mice.Thirty-six mature Sprague-Dawly mice were divided into 2 groups at random.CS was transplanted into the left(group A 1,n=9) and ADBM into the right(group A2,n=9) thigh muscle pouches.HDBM was transplanted into the left thigh muscle pouches(group B 1,n=9) and the right sites were taken as blank controls(group B2,n=9).Experiments were done to induce ectopic bone formation.At 2,4,6 weeks postoperatively,specimen were collected to evaluate gross and tissue structures and biochemical tests for alkaline phosphatase(ALP) and Ca2+ so that osteoinductive activities of different bone graft materials could be assessed.[Result]At 2 weeks postoperativly,ADBM and HDBM were wrapped up by fibrous tissues and stromal cells gathering around the DBM slices.At 4 weeks postoperativly,formation of cartilage and osteoblasts were observed,and at 6 weeks,materials like cartilage matrix were observed to grow.The concentration of ALP and Ca2+ in study groups was higher than that of control group,which meant that 2 types of DBM had osteogenic potential and that the differences of osteogenetie potential in ADBM and HDBM relied on the donors,whereas CS could be degraded and absorbed fast with light inflammatory reaction and no ectopic bone formation was observed in CS graft.[Conclusion]Both ADBM and HDBM have osteoinductive potential.ADBM is better in inducing ectopic bone formation than HDBM.Differences in donors and preparation of ADBM and HDBM have impacts on their abilities of inducing ectopic bone formation.CS is good at biocompatibility and could be used as bulking agents to repair bone defects.

Objective To study the experimental protocol for purification and analysis of hematopoietic stem cells(HSC)and myeloid lineage-committed progenitors.Methods According to differentiation antigen expression pattern on hematopoietic stem cells(HSC) and progenitors during hematopoietic development,HSC and progenitors from bone marrow of 14 healthy mice were analyzed and sorted by magnetic nanoparticles and 4-color or 6-color flow cytometry using multiple antibody panels.Sorted HSC and progenitors were further tested by methylcellulose colony forming unit(CFU)and serial replatingassays.Results The expression of hematopoietic progenitor cells(HPC)was 10-fold higher expression than that of HSC.However,replating activity of common myeloid rogenitors(CMP)was only half of that of HSC.And there was almost 120 replating activity observed in granulocyte/macrophage lineage-restricted progenitors(GMP)and megakaryocyte/erythroeyte lineage-restricted progenitors(MEP).Conclusion Multiparametric flow cytometry could be used to isolate and count HSC and myeloid lineage-committed progenitors accurately.

Objective To investigate the mutations of basal core promoter (BCP) and precore (PreC) region of hepatitis B virus (HBV) and the association with the development of hepatocellular carcinoma in patients with chronic HBV infection.Methods Totally 381 untreated HBV patients were recruited from the Department of Infectious Diseases,People's Hospital of Shangyu from Jan 2003 to Dec 2010,which included patients with chronic hepatitis B (CHB,n =166),cirrhotic hepatocellular carcinoma (cirrhotic-HCC,n =158) and noncirrhotic hepatocellular carcinoma (noncirrhotic-HCC,n=57).The mutations in HBV BCP and PreC and the genotypes of HBV were determined by polymerase chain reaction (PCR) and direct sequencing.Data were analyzed by chi square test and Logistic regression.Results The HBV genotype of most cases was genotype B (CHB,n =124;cirrhotic-HCC,n=126 ; noncirrhotic-HCC,n=50).In univariant analysis,BCP V1753 (x2 =7.927,P=0.005),BCP T1762/A1764 (x2 =12.796,P＜0.01),PreC A1896 (x2 =6.890,P=0.009) and PreC A1899 (x2=11.850,P =0.001) mutations were more frequently detected in cirrhotic-HCC patients than those in CHB patients.PreC A1896 (x2 =27.310,P＜0.01) and A1899 (x2=7.575,P=0.006) mutations were highly detected in noncirrhotic-HCC patients than those in CHB patients.Multivariate Logistic regression analysis revealed that in HBeAg positive patients,BCP T1762/A1764 (wald=6.180,P=0.016,OR=8.883) and PreC A1899 (wald=10.279,P=0.001,OR=7.475) mutations were independently associated with the development of cirrhotic-HCC; PreC A1896 (wald=4.324,P=0.038,OR=4.439) and PreC A1899 (wald=4.850,P=0.028,OR=6.010)mutations were independently associated with the development of noncirrhotic-HCC.While in HBeAg negative patients,PreC A1896 mutation (wald=15.448,P＜0.01,OR=12.128) was independently associated with the development of noncirrhotic-HCC.Conclusions BCP T1762/A1764 mutations are associated with the development of cirrhotic-HCC in HBeAg positive patients.PreC A1896 mutation is associated with the development of noncirrhotic-HCC in HBeAg positive and HBeAg negative patients.PreC A1899 mutation is associated with the development of cirrhotic-HCC and noncirrhotic-HCC in HBeAg positive patients.

Objective To value the effect and safety of butyl phthalide sequential treatment in cerebral infarction of branch sclerosis of arterial congee appearance. Methods Sixty patients with acute cerebral infarction were randomly divided into treated group( n = 31)and control group( n = 29). According to the condition of illness,all patients were given aspirin,atorvastatin calcium,and the injection of ozagrel sodium intravenous;controlled the blood pressure,blood sugar,blood lipid,and treated complications posstively;take the early rehabilitation of nerve treatment afte the illness was in stable condition. Butyl phthalide was used in the patients of treated group(100 ml,twice per day,intravenous drip,during 14 days period therapy,and then 0. 2 g oral,third per day),besides the routine therapy. The degree of neural function defect score( NIHSS)and activities of daily living score(BI)between two groups were observed before and after treatment. Corresponding adverse consequences were recorded. Results Compared with pretreatment,the NIHSS of postreatment at the 14th day in treat and control groups were decreased(treated group:(4. 36 ± 3. 11)vs.(11. 42 ± 3. 20);control group:(6. 12 ± 2. 67)vs.(11. 64 ± 3. 43),P < 0. 05,and the treated group was significantly lower than control group(F inner groups = 2. 125,P < 0. 01;F between groups = 18. 63,P < 0. 01;F cross groups = 25. 34,P< 0. 01;P < 0. 05). The BI of postreatment in two group were increased(treated group:(86. 72 ± 8. 44)vs. (26. 54 ± 13. 36);control group:(75. 96 ± 9. 86)vs.(26. 38 ± 13. 02)),and the treated group was significantly lower than control group(F inner groups = 29. 27,P < 0. 01;F between groups = 32. 48,P < 0. 01;F cross groups= 42. 41,P < 0. 01;P < 0. 05). There was no the adverse reactions. Conclusion Butyl phthalide sequential treatment can improve the NIHSS and BI of cerebral infarction of branch sclerosis of arterial congee appearance and have a better therapy effect.

Objective To establish a capillary gas chromatography method for determination of camphol and isoborneol in Shaoshang yuhe gao ( burn healing cream) . Methods The capillary gas chromatography was adopted under the following conditions: use PEG-2000 as the stationary liquid,nitrogen as carrier gas,ZB-WAX (30 m×0. 25 mm,0. 25 μm) as the chromatographic column,and the flame ionization detector. The column temperature was programmed at 80 ℃ for 5 min as the initial temperature,then raised to 180 ℃ at the rate of 5℃·min-1 and kept for 10 min. The shunt ratio was 101. Results The liner range for camphol was 0. 487 5-31. 25 μg ( r =0. 999 6),and the average recovery was 95. 95%( n =6). The liner range for isoborneol was 0. 487 5-31. 25 μg( r =0. 999 7),and the average recovery was 96. 44%( n =6). Conclusion The method is accurate,sensitive,and can be applied to quality control of shaoshang yuhe gao.

Objective To compare the pre-existing mutations in reverse transcription region of HBV in patients with different HBV infection stages.Methods Totally 474 patients with chronic HBV infections,including 205 with chronic hepatitis B (CHB),153 with liver cirrhosis and 116 with hepatocellular carcinoma (HCC),were enrolled from the People' s Hospital of Shangyu and the First Affiliated Hospital of Zhejiang University during January 2011 and June 2013.All patients had not received nucleos (t)ide analogues treatment.HBV RT region mutations and genotypes were determined by PCR followed by sequencing.SPSS14.0 was used for statistical analysis.Results There were 387 (81.6％) patients with HBV genotype B,in which 156 were with CHB,124 were with liver cirrhosis,and 107 were with HCC.Nucleos(t)ide analogues-related mutations were observed in all the above 387 patients.rtS106C mutation was more popular in CHB and liver cirrhosis (14.1％ and 14.5％) patients than that in patients with HCC (4.7％) (x2 =6.126,6.207,P ＜0.05); And the positive rates of rtD134E/G/N/S mutations were also higher in CHB and cirrhotic patients (21.8％ and 20.2％) than that in HCC patients (10.3％,x2 =5.933,4.263,P ＜ 0.05).rtD134E/G/N/S and rtS106C mutations were correlated with HBeAg (P ＜0.01) and gender (P ＜ 0.05),but not with HBV virus load and age (P ＞ 0.05).The mutation frequencies in A-B interdomain were higher in CHB and cirrhotic patients (5.3％ and 5.6％) than that in HCC patients (3.5％,x2 =9.018,11.018,P ＜ 0.01).Conclusions Nucleos (t) ide analogues-related mutations exist in various HBV infection stages.rtSl06C and rtD134E/G/N/S mutations may be involved in necro-inflammation,and A-B interdomain mutations may be correlated with necro-inflammation,immune response and fibrosis in chronic liver diseases.

Objective To evaluate the radiation method and resuh of 450 patients received TBI(total body irradiation).Methods Single-dose Measurement was used to mark dose of TLD(thermo luminescence dosimeter).The values of actual dose in body midline were evaluated by calculating and correcting mean dose of incidence and emergence.Radiation methods:In four-field Irradiation.diagonals of fields coinside with the longitudinal axis of the patients,patient in supine and lateral positions received two pairs of parallel opposite radiation.Scheme of TBI came from a preparative radiation about one week before,and this four-field and equal-in-dose(about 10%of TBI)preparative radiation offered US the optimal scheme with aminimal dose non-uniformity by adjusting different dose proportion of supine and lateral position.In small field irradiation,patients received one pair of parallel opposite radiation from lateral side sitting on a special stool with backrest,the stool can be rotated CW or CCW,pedals can be move forward or backward and fixed.In opposite lateral irradiation,similar to four-field irradiation,patients received one pair of horizontal opposite radiation only in supine position.Five of these patients received FTBI(Fractional TBI). Results The average non-uniformity in midline of patients in four-field irradiation group(87 patients).small field irradiation group(91patients)and opposite lateral irradiation group(272 patients)is respectively ±8.1%,±7.4% and ±4.9%. Conclusions It iS a important process for QA and Qc to measure the dose of incidence and emergence real-timely with TLD or semiconductor dosimeter.We can adopt small field irradiation when the field iS not large enough to contain the patient from head to foot,and it showed advantages over four-field irradiation in treatment process and outcomes.We found the uniformity in body midline would be much better in supine position with diagonal＞180 cm than that in four-field irradiation and small field irradiation with diagonal＜110 cm.We compared supine position irradiation with opposite lateral irradiation,only to find which has its strong point.And actually we considered that FTBI treatment booth can be used more often in anterior and posterior parallel fields irradiation,patient semi-sitted,repeatedly received forward and backward radiation. In spit of not possessing radio-biological advantages as FTBI,STBI(Single TBI)is still a practical form of TBI.