An immunosuppressive early pregnancy factor (EPF) present in the serum of pregnent women, between 3 and 6 weeks of gestation, was isolated and purified using ion-exchange chromatography, affinity chromatography and gel filtration techniq(?) The serum samples and chromatographic fractions were measured for EPF activity using rossette inhibition test and hCG content by radioimmunoassay. The result showed that the column eluates (?) gel filtration exhibited no hCG (3. 1 ng/ ml). The fraction containing the bulk of EPF on SDS polyacrylamide gel electrophoresis revealed the presence of polypeptide bands of molecular weight at 24 kDa and 41 kDa respectively.

Clinical efficacy of liver lesion with the treatment of reduced glutathione and ademetionine was analyzed retrospectively. 83 patients were randomly divided into two groups based on the application of preventive hepatopro-tective drug. Control group was treated with reduced glutathione intravenous drip infusion once a day ( n =40 ) , while treatment group with reduced glutathione and ademetionine(Transmetil) once a day(n=43). After 12 days, the clinical efficacy of treatment group was better than that of control group. Total response rate was 95. 35% for treatment group, much better than that of control group(80. 00%). There was significant difference between two groups ( P<0.05 ) . Reduced glutathione and ademetionine are more effective in the treatment of chemotherapeutics-induced liver lesion than only with reduced glutathione.

Objective To study imaging features of neuronal tumors, and to evaluate the value of CT and MRI in diagnosing these tumors. Methods The CT and MRI studies in 10 patients with pathologically proved neuronal tumor were retrospectively reviewed. Results (1) CT showed a mixed attenuation cystic solid mass with calcification in 3 gangliogliomas; The mixed signal intensity on T 1 or T 2WI with heterogeneous contrast enhancement was common in ganglioglioma. (2)Three central neurocytomas had mixed signal intensity. (3)Three dysembryoplastic neuroepithelial tumor (DNT) had decreased signal intensity on T 1 weighted MR images and well demarcated increased signal intensity on T 2 weighted images without peritumoral edema and calcification. Prominent MR imaging features were a gyriform configuration on T 1 or T 2WI. The tumor contrast enhancement was not common; (4)A desmoplastic infantile ganglioglioma (DIG) was located in the frontal and parietal lobes, appearing as cystic tumors with a solid cortical component enhanced intensely. Conclusion Some neuronal tumors possess of specific imaging features, but diagnosis of the tumor with imaging modalities alone may be difficult. Therefore, combined clinical history could help to establish a correct diagnosis.

Objective To construct shifting mutant of cysteine-rich region to 3?@terminal of Tat gene of human immunodeficiency virus type-1 (HIV-1) HXB2 strain, and to analyze the immunogenicity of mutant protein (Tat-cct) after prokaryotically expressed and purified. Methods The cysteine-rich region (nucleotides 64--111) of Tat gene was shifted to 3'terminal of Tat of HIV-1 HXB2 strain by polymerase chain reaction (PCR) and Tat mutant DNA sequence was obtained. Prokaryotie express plasmid pET32a-Tat-cct was constructed and transformed into E. coli BL21 (DE3), then Tat-cct protein was expressed and purified. BALB/c mice were immunized with the fusion protein Tat-cct, and immunogenicity of the immunized serum was detected by enzyme-linked immunosorbent assay (ELISA). Results The recombinant plasmid pET32a-Tat-cct expressed in E. coli BL21 (DE3) and the relative molecular mass of the purified fusion protein was 31 000. The serum antibody titer of mice immunized with Tat-cct recombinant protein was 1 : 1600, which binded specifically with both Tat-ect protein and Tat protein (amino acids 1-101). Conclusions The recombinant protein Tat-cct of Tat mutant strain can be expressed efficiently in E. coli and well retains immunogenicity, which provides valuable information for basic research of HIV-1 Tat vaccine.

Objective Deleting the cysteine-rich region (22-37 amino acids)of HIV-1 HXB2 Tat protein(whole length is 101 amino acids) to improve its stability and expression level in E.coli and to analyze the immunogenicity of Tat protein without the cystein-rich region [Tat(△C)protein]. Methods Tat DNA deleted the cysteine-rich region (64-111 nucleotides), named as Tat(△C)DNA, was obtained in vitro by PCR and cloned into pET-32a vector. pET-32a-Tat(△C)plasmid and the pET-32a-Tat plasmid were established and transformed into E.coli BL21(DE3) strains respectively to express and purify the protein. Three rabbits were vaccinated with pET-32a-Tat(△C)protein, then testify the reactivity of sera from rabbits by ELISA and Western blot. Results The dense of the purified pET-32a-Tat(△C)protein was 7.12 mg/ml,which was greatly more than pET-32a-Tat protein(1.50 mg/ml). Dimer of pET-32a-Tat protein can be observed just after the protein purification and stored at 25℃ and 4℃ for 7 days, but dimer of pET-32a-Tat(△C)protein was not formed at the same condition. Experimental rabbits were immunized with pET-32a-Tat(△C)protein and produced high titre of anti-pET-32a-Tat(△C)serum(1∶320 000), the antibody can react specifically with Tat(△C)protein, Tat protein (1-101 AA)and synthetic Tat(1-86 AA) protein. Deletion mutation of the cysteine-rich region of Tat protein was first performed in the study. Conclusion The expression level in E.coli and the stability of Tat protein deleted the cysteine-rich region can be increased greatly, and the protein remains good immunogenicity. The results may provide a novel antigen for further development of HIV-1 Tat vaccine.

We constructed a phage-displayed random mutation library of Tat38-61(51N/55N), for studying the molecular evolution screening of HIV-1 Tat38-61 epitope. We used primers containing the random nucleotide sequences, and introduced the random mutations at the sites of 51 and 55 amino acids coding sequences into full-length Tat sequences by overlapping PCR. With the randomly mutated full-length Tat as template, the Tat38-61(51N/55N) mutants which contained recognition sequences for the Xba I in both ends were amplified by PCR using the designed primers. The mutants were cloned into Xba I site in the phagemid vector pCANTAB5S, then the recombinants were transformed into E. coli TG1, a phage-displayed the random mutation library of Tat38-61(51N/55N) was constructed by the rescue of help virus M13KO7. The results showed that the library consisted of about 5.0 x 10(6) colonies and the phage library titer was 2.65 x 10(12) TU/mL. More than 56.50% colonies in the library were positive for insertion. Sequence analysis showed that the nucleotides encoding amino acids at the sites of 51 and 55 distributed randomly. The constructed mutation library could meet the requirements for the following molecular evolution screening, and might prepare the Tat mutants for the further study of new Tat vaccine candidates.
AIDS Vaccines ; immunology ; Escherichia coli ; genetics ; metabolism ; HIV-1 ; genetics ; Humans ; Mutation ; Peptide Fragments ; biosynthesis ; genetics ; immunology ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; tat Gene Products, Human Immunodeficiency Virus ; biosynthesis ; genetics ; immunology

OBJECTIVETo evaluate the difference in the clinical therapeutic effects on anal fissure at Ⅰ and Ⅱ stages between the acupoint catgut embedding therapy and western medication.

METHODSSixty patients of anal fissure at Ⅰ and Ⅱ stages were randomized into an embedding therapy group and a western medication group, 30 cases in each one. In the embedding therapy group, the acupoint catgut embedding therapy was applied at bilateral Tianshu (ST 25), Changqiang (GV 1), bilateral Chengshan (BL 57) and Tigangxue (Extra), once a week. In the western medication group, the external inunctum on the wound was given with 0.2% nitroglycerin ointment, once every morning and evening a day. The treatment lasted for 4 weeks continuously in the two groups. The follow-up visit was done for 3 months after treatment. The visual analogue scale (VAS) and anal pain duration were observed and recorded before treatment and on the 3rd day and the 7th day of treatment separately. The clinical therapeutic effects were compared between the two groups.

RESULTSAfter treatment, on the 3rd day and the 7th day of treatment, VAS score and anal pain duration were all reduced significantly as compared with those before treatment in the patients of the two groups (all<0.01). The differences in the embedding therapy gruop were better than those in the western medication group before and after treatment (<0.01,<0.05). In the 2nd and 4th weeks after treatment, the clinical therapeutic effects in the embedding therapy group were better than those in the western medication group (both<0.05). In 3-month follow-up, the recurrent case in the embedding therapy group was one, and the recurrent case in the western medication group was six.

CONCLUSIONSThe acupoint catgut embedding therapy is safe and effective in the treatment of anal fissure at Ⅰ and Ⅱ stages and its recurrent case is lower as compared with the treatment of western medication.