Objective To investigate the clinical significance of dynamic changes of CD4~+ CD25~+ regulatory T cells(Treg)in patients subject to allogeneic hematopoietic stem cell transplanta- tion(alIo-HSCT).Methods Forty-five patients received allo-HSCT.The graft-vesus-host disease (GVHD)was prevented by cyclosporine A and short-term MTX regimen in 31 patients.Fourteen of all the patients received Zenapox(CD25MAb)at the day of transplantation and day 4 after transplan- tation.The levels of Treg in peripheral blood were detected by flow cytometry from 45 patients at 2nd,4th,8th and 12th week after allo-HSCT and the time of aGVHD development,respectively.Re- suits Anti-CD25 could suppress the peripheral blood levels of Treg significantly.The Treg levels were significantly higher in patients with grade 0-1 aGVHD than those with 2-4 aGVHD at 8th and 12th week after transplantation.Among patients with 2-4 aGVHD,Treg levels were significantly low- er after development of aGVHD than before.Conclusions Treg are important for the aGVHD preven- tion and can be a useful clinical surveillant index for the development of aGVHD.It can significantly decrease the levels of Treg in the peripheral blood with anti-CD25.

Objective To screen proteins binding with interferon?(IFN?)from human hepatic cDNA libraty by yeast-two hybrid technique.Methods The IFN?gene was amplified by polymerase chain reaction(PCR)and constructed into pGBKT7 vector as the bait plasmid in yeast-two hybrid system3,pGBKT7-IFN?was then transfected into yeast AH109.The transfected yeast were mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium.Diploid yeast was plated on synthetic dropout nutrient medium(SD/-Trp Leu-His-Ade)and synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)containing X-?-gal for selecting.After plasmid extracting and en- zyme cutting analysis,the blue colonies were performed sequence analysis,the results were analyzed by bioinformatics.Results IFN?gene was successfully cloned and expressed in yeast cells.Thirty- four positive colonies were obtained using yeast-two hybrid technique.After sequence analysis,eight clones were found may have a binding effect with IFN protein.Conclusions IFN?genes was success- ful cloned and eight proteins that could bind with IFN?protein were also screened.

Objective To explore the dose-effect relationship between water fluoride levels and hepatic damage in children and observe the difference in hepatic function between high-loaded fluoride people and dental fluorosis people in the same water fluoride level region. Methods 210 children were selected and divided into seven groups according to drinking water fluoride concentrations and whether they suffered from dental fluorosis. Urine and serum fluoride content total protein TP and albumin ALB content and activities of ALT AST and LDH in serum were determined. Results Both of urine and serum fluoride of high fluoride people and dental fluorosis people were higher than those of the control moreover fluoride contents in urine and serum increased gradually with the increase of fluoride level in drinking water. No significant differences were seen in serum TP ALB ALT and AST levels among groups. Serum LDH activities significantly increased in dental fluorosis people from area of 2.58 mg/L fluoride in drinking water and in two groups from area of 4.51 mg/L fluoride. Moreover there was an obvious dose-effect relationship between the drinking water fluoride concentration and LDH activity. Conclusion If the concentration of fluoride in drinking water exceed 2.0 mg/L it will cause hepatic damage in children with an remarkable dose-effect relationship. The degree of hepatic damage is related to not only water fluoride level but also the condition with or without dental fluorosis.

Objective To explore the expression and significance of hedgehog signal molecules (Ptch,Smo and Gli1 ) in chronic pancreatitis tissues in rats.MethodsSixty SD rats were randomly divided into CP group (n =50) and control group (n =10).DBTC solvent (8 mg · ml-1 · kg-1 ) was injected into the rat via tall vein in CP group.In control group,rats were treated only with the solvent at a dose of 1ml/kg body weight.All rats were sacrificed 6 weeks later to observe the pancreatic pathologic changes.Collagen accumulation in pancreatic sections was determined by staining for Sirius red.Expressions of Ptch,Smo,Gli1 mRNA and protein in pancreatic tissues were assessed by RT-PCR and immunohistochemistry.ResultsThe rate of chronic pancreatitis development in rats in CP group within six weeks was 73.9％.Collagen content was markedly higher in CP group than that in control group [ ( 38.52 ± 6.49 ) ％ ~s (7.37 ± 2.28 ) ％,P ＜ 0.05 ].No Path,Smo,Gli1 protein expression was observed in normal pancreatic tissues in control group.The positive rate of Ptch,Smo,Gli 1 expression was 73.5％,64.7％ and 52.9％ in CP group,and the difference between the two groups was statistically significant (P ＜ 0.05).The expressions of Ptch,Smo,Gli1 mRNA were 2.38 ±0.42,3.85 ± 1.03,4.63 ± 1.49 in CP group,which were significantly higher than those in control group (0.23 ±0.16,0.14 ±0.05,0.57 ±0.12,P ＜0.05).ConclusionsThe Ptch,Smo,Gli1 was highly expressed in pancreatic tissues in CP rats,suggests hedgehog messenger pathway may play an important role in the chronic inflammation and fibrosis of chronic pancreatitis.

Adult ; Causality ; Cross-Sectional Studies ; Efficiency ; Female ; Humans ; Male ; Middle Aged ; Occupations ; Social Support ; Stress, Psychological ; Surveys and Questionnaires

Child, Preschool ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gastrointestinal Diseases ; drug therapy ; Humans ; Infant ; Male ; Phytotherapy ; Plant Extracts ; therapeutic use

The mainly antigenic sites for the adenovirus neutraliation are present on Loop1 and Loop2 of hexon.Majority research were focus in the human adenovirus.Little was known on infectious canine hepatitis virus (ICHV), which was also called canine adenovirus typeⅠ.Here,ICHV (the isolated strain) DNA was isolated and purified from the cultured MDCK cells.The Loop1 and Loop2 fragments were amplified by polymerase chain reaction(PCR) method,and then was connected by ligase T4.The target fragment was then connected with vector pET28a.The nucleotide sequence ecoding Loop1 and Loop2 was determined.The nucleotide sequence identity of Loop1 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 100%, 100% and 83.8%, and the nucleotide sequence identity of Loop2 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 88.1% , 88.1% and 99.3%, and amino acid identity is 93.6%, 93.6% and 98.6%.The recombinant Loop protein was expressed in E.coli and was approximately 36kDa in size,and then was purified. Then BALB/c mice were injected subcutaneously in the back and armpit with the recombinant Loop protein.The anti-ICHV antibody titers of immunized serum was tested by indirect ELISA and the titers were up to 1:320.Western blot demonstrated that immunized sera could specifically combine with ICHV. The research laid a foundation for creating new genetic engineering products of infectious canine hepatitis virus.

Bicyclol with benzyl alcohol structure, is a poorly water-soluble drug, used for the treatment of chronic hepatitis B. To increase the drug solubility and oral bioavailability, a Bicyclol-phospholipid complex was studied on its preparation, formation mechanism, and the influence on drug physicochemical properties and oral absorption. The complex was prepared by a solvent evaporation method. The optimal formulation was selected by orthogonal experimental design, and a reasonable evaluating method of the complexation rate was established. Various methods, such as differential scanning calorimetry (DSC), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and 31P nuclear magnetic resonance (31P-NMR), were used to explore the phase state and formation mechanism of the complex. The solubility of drug in complex was investigated in water/n-octanol. Preliminary study of its absorption and liver tissue distribution in rats was also carried out. The results showed that Bicyclol and phosphatidylcholine can be complexed entirely in the molar ratio 1 : 2. Bicyclol was dispersed in phospholipids as amorphous state. They were combined by intermolecular hydrogen bond due to charge transfer effect which occurred between the two polarities of the double bond between phosphorus and oxygen (P=O) of phosphatidylcholine and benzalcohol group of Bicyclol. The solubility of the complex compared to the active pharmaceutical ingredient (API) was effectively enhanced 5.75 times in water and 7.72 times in n-octanol, separately. In addition, drug concentrations were also enhanced 43 times in plasma and 13 times in liver with one hour after administering the complex to rats via oral gavage. All of these indicated that Bicyclol with benzalcohol group can interact with phospholipids to form complex, improving drug's physicochemical properties, thus further increasing its absorption and target tissue distribution. This study also provided theoretical reference for the research of other benzalcohol derivatives complexed with phospholipids.
1-Octanol ; Animals ; Biological Availability ; Biphenyl Compounds ; pharmacokinetics ; Calorimetry, Differential Scanning ; Chemistry, Pharmaceutical ; Phospholipids ; pharmacokinetics ; Rats ; Solubility ; Spectroscopy, Fourier Transform Infrared ; Tissue Distribution ; X-Ray Diffraction

Injectable lipid emulsions have been routinely used in patients since 1960s as a nutritional supplement for patients requiring parenteral nutrition. In recent years, lipid injectable emulsions have been extensively studied as a kind of novel drug carrier, also the quality problems of the lipid emulsion attract more and more attentions gradually. Large diameter tail of injectable lipid emulsions as a significant quality control indicator should pay more attention. Regarding to the defect of detecting large diameter tail of lipid injectable emulsions in our country, the purpose of this article is to summarize the techniques of detecting large diameter tail, illustrate the impacts of large lipid droplet on the quality of lipid injectable emulsions, emphasize the importance of detecting large diameter tail in lipid emulsions and provide guidance for researching and developing lipid emulsions in domestic market.
Drug Stability ; Fat Emulsions, Intravenous ; chemistry ; Lipids ; chemistry ; Parenteral Nutrition Solutions ; chemistry ; Particle Size ; Quality Control

The aim of this research is to refine the protocol of purification of SA and identify the character of SA. By utilizing the cold-denaturing method, most of other kinds of protein were screened out and SA was purified from the fermentation broth of L-183 by using the refined affinity chromatography method. The rate of recollection was checked to be 75%～85%. By identification, it is indicated that the molecular weight of self-made SA was 74.5kD, the biotin-combining number 3.2, the activity 11.2u/mg, the pI around 7.4. So, the essential characters of SA are same as described by documents.