Objective To detect whether trichostatin(TSA) can induce HL-60 differentiation in vitro. Methods MTT method was used to test the effect of TSA on HL-60 cell growth. Cell cycle was tested by flow cytometry. CD11b expression was tested for detecting cell differentiation, RT-PCR was used to detect the mRNA expression of c-myc in the cells treated by TSA. Results Down-regulation of cell proliferation was observed and cells significantly accumulated at the G0 and G1 phase in HL-60 cells treated with TSA( P <0. 01). Dif-ferentiation rate was 15. 24% after being treated by TSA for 48 h. mRNA of c-myc was down regulated in time-dependent manner. Conclusion TSA can inhibit proliferation and induce differentiation in HL-60 cells.

Human fetal esophageal epithelial tissue were cultured in vitro and treatedwith mycotoxins of Alternaria alternata (AME or AOH) for 4 h. The genomic DNAwere extracted from these tissues. Genomic DNA was isolated from normal human fetalesophageal epithelium (as blank control), DNA from malignant tissue and its adjacentnormal mucosa was obtained from esophagectomy patients. DNA was amplified with PCRreaction, using genomic DNA as templet. The PCR products was a 104bp fragment from which the 12 codon of c-Ha-ras gene was contained. The excition point of restriction en-zyme Hpe Ⅱ was located in this fragment. The PCR amplified 104bp fragment was diges-ted by Hpa Ⅱ and analysed by agarose gel electrophoresis. The results showed that the104bp fragment amplified from genomic DNA of blank control and esophagectomy patientcould be digested by Hpa Ⅱ ; but that from genomic DNA of human fetal esophagealepithelium treated by AME or AOH could not. These results indicated that a mutationhad taken place at 12-codon of c-Ha-ras gene after it was treated by AME, AOH for ashort time. The mutation of Ha-ras gene might be the early event during esophageal car-cinogenesis. The effect of AME and AOH during the onset of esophageal cancer and themolecular machanisms of the effect were worth of further study.

Objective To investigate the mechanism of acquisition of HLA-G1 antigen by NK cells.Methods K562 cells stably expressing HLA-G1 antigen (K562-G1) were constructed.K562-G1 cells, K562 cells and shed HLA-G1 were respectively co-cultured with NK-92MI cells to observe the acquisi-tion of HLA-G by NK cells.To further investigate the mechanism , NK-92MI cells with blockage HLA-G re-ceptors were further co-cultured with K562-G1 cells and HLA-G1 proteins expressing on K 562-G1 cells were blocked and then co-cultured with NK-92MI cells. Acquisition of HLA-G 1 by NK-92MI cells was analyzed by flow cytometry and fluorescence microscopy .The effects of HLA-G1 expression on the cytotoxicity of NK-92MI cell were evaluated by flow cytometry analysis based on CD 107a labeling.R esults NK-92MI cells could quickly acquire HLA-G1 from K562-G1 cells in co-culture experiments .Blockade of HLA-G1 or its re-ceptors KIR2DL4 and ILT2 with specific mAbs did not affect the acquisition of HLA-G1 by NK-92MI cells. Moreover, HLA-G1 could significantly inhibit the cytotoxicity of NK cell ( P<0.01).Conclu sion NK-92MI cells acquire HLA-G1 from K562-G1 cells via trogocytosis , which is not associated with affinity be-tween receptor and ligand , extracellular domain of HLA-G1 or passive adhesion .

Objective To establish the expression of membrane-bound HLA-G1-G4 isoforms in choriocarcinoma cell line JAR and to investigate its roles in NK cytotoxicity in vitro. Methods Stable expression of HLA-G1, -G2, -G3 and -G4 in JAR cells was established by gene cloning and transfection.HLA-Gtranscripts and protein expression in the transfected JAR cells was tested by RT-PCR, flow cytometry, Western blot and immunocytochemistry, respectively. High-affinity peptide KIPAQFYIL pulsing was performed to evaluate its effects on HLA-G expression. Effects of HLA-G1-G4 isoforms on NK cytotoxicity was performed with lactic dehydrogenase (LDH) releasing method. Results RT-PCR, Western blot and immunocytochemistry results showed that exogenous HLA-G1-G4 gene were successfully transfected and proteins were stably expressed in the HLA-G negative JAR cells; Flow cytometry data showed that only HLAG1, but not HLA-G2-G4 isoform was detectable in those transfected JAR cells and the peptide pulsing did not affect their expression status. However, all HLA-G1-G4 isoform expressed JAR cells could significantly decreased the NK cell cytotoxicity (P＜0.05). Conclusion HLA-G1-G4 isoform expression could dramatically inhibit NK-92 cell lysis, indicating that membrane-bound HLA-G isoforms are importantly immunotolerant and may play immune regulation roles in various physio-pathological situations.

[Objective]We summarized Professor Zhang Weihua’s clinical experience in treating constipation and diarrhea alternating bowel dysfunction by Chinese medicine. [Method] This paper revealed the professor Zhang Weihua ’s experience of curing constipation and diarrhea alternating bowel dysfunc-tion by analyzing the etiology and pathology ,therapeutic principles and relevant cases .[Results] The etiology and pathology of constipation and diarrhea alternating bowel dysfunction is root deficiency and tip excess ,and cold-heat complex ,the disorders of E. conduction,and the treatment should focus on using warming and heat-clearing simultaneously, tonifying spleen and kidney. [Conclusions] The experience of Professor Zhang Weihua in treating con-stipation and diarrhea alternating bowel dysfunction is effective and thought-provoking in treatment.

Objective To discuss the significance of stress roentgenogram on diagnosis of chronic instability of lateral ankle and. bring forward the radiology diagnosis criteria. Methods 40 patients with chronic instability of lateral ankle and 40 normal people were randomly selected. Bilateral ankles of each subject had two basic roentgenographic measurements named inversion stress anteroposterior roentgenogram and anterior drawer stress radiograph. The talar tilt angel and anterior translation of talus were measured. Results The average TT and ATT of suffered ankles are 9.1 and 7.8mm,while the values of the opposite ankles are 5.4 and 5.4mm, the comparison group are 4.9 and 6.1mm.There is significant difference between the suffered ankles and normal ones(P

Objective To investigate the association between the systolic/diastolic orthostatic hypotension (OH-S/OH-D) and myoeardial infarction (MI) in the elderly.Methods Health screening physical examination were carried in 1081 subjects without MI aged over 65 years in Guangzhou Military region.The orthostatic blood pressure and heart rate were measured in supine position after resting for more than 5 minutes and at 0 and 2 minutes after standing.All the cases were divided into systolic or diastolic group on the basis of definition of orthostatic hypotension and followed up by telephone or inpatient medical records with mean period of 315.8 days.The primary endpoint was MI occurrence.Results The prevalence of OH in this cohort was 24.5％ ( OH-S/OH-D:19.3％/17.2％ ).Significant differences in the occurrence of OH and OH-S were found in the elderly and the very elderly subjects( ≥80 years) (26.1％ vs 20.1％,P=0.045; 21.0％ vs 14.6％,P =0.018),while no difference was found in OH-D.The prevalence of MI in the OH positive subjects was significantly higher than that in the OH negative subjects,as well as in OH-S or OH-D group.After adjustment of age,supine blood pressure,creatinine and cerebrovascular history by logistic regression,the association was observed between MI and OH ( HR 15.635,95 ％ CI 3.299-74.091,P=0.001),OH-S(HR 8.760,95％CI2.487-30.851,P=0.001)and OH-D(HR 3.889,95％ CI 1.097-13.790,P =0.035 ).Conclusion OH-S and OH-D hypotension are robust predictors for MI in the elderly.

Objective To investigate the mechanism of different HLA-G isoform mRNA patterns in different cells alters its cell membrane expression.Methods RT-PCR was used to analyze HLA-G isoform mRNA(HLA-Gl-6)of ovarian cancer cell lines HO-8910,HO-8910PM and OVCAR-3,leukemia cell lines Jurkat,K562,HIJ60,MUTIZ-1,and the chorioeareinoma cell lines JEG-3,JAR.HLA-G between cellular membrane and intracellular expression were analyzed by flow cytometry.Results All HLA-G mRNA isoforms were observed in the positive control cell line JEG-3,but none in the negative control cell line JAR.HLA-G1 isoform mRNA was expressed in HO-8910,HO-8910PM,OVCAR-3,MUTZ-1 and Jurkat cells.HLA-G2 mRNA was not detected in any cell line but JEG-3.HLA-G3 mRNA was found in HO-8910,HO-8910PM,K562,HIJ60,MUTZ-1,OVCAR-3 and Jurkat cells.HO-8910,HO-8910PM,HIJ60,Jurkat cells expre8sed HLA-G4 mRNA.Only the Jurkat cells expressed HLA-G5 mRNA.FACS results showed that JEG3 and HO-8910PM cells membrane expressed HLA-G,however,the intraeellular HLA-G expression was detected in all tested cells except the negative control cell JAR.Conclusion Only the HLA-G1 isoform could be exDressed on cell membrane in particular cell lines. Other isoforms including HLA-C2,-G3,-G4,-G5 and HLA-G6 could not reach cell snrface.

Objective To explore the effect of oris(oral)muscles training on speech language rehabilitation for autism spectrum disorders children.Methods 40 cases were divided into 2 groups,20 cases in each group.The control group was treated with language cognitive training.The observation group was treated with language cognitive combined with oris(oral)muscles training.The treatment was given for 6 months.S -S(sign -significate relationgs) was used to evaluate the language development quotient of the two groups.Results After treatment,the language development quotient was significantly improved compared with before treatment(P <0.05),and the language expres-sion quotient of the observation group was more effective than the control group(t =2.434,P <0.05).The language comprehension quotient and operation quotient of the two groups had no significant differences.Conclusion Oris muscles training combined with language cognitive is more effective on language expression capability of autism spec-trum disorders children.

Objective To explore the treatment of the ICU ventilator associated pneumonia(VAP),and the prevention strategy.Methods Analyzed the clinical features and microbiological data of 80 ICU VAP patients. Results The main pathogenic bacterias of VAP were acinetobacter baumannii and pseudomonas aeruginosa,which were Gram negative bacteria.A total of 200 strains pathogenic bacteria were isolated,in which,Gram negative bacteria reached to 76.0%,which mainly including:acinetobacter baumannii strains,klebsiella pneumoniae,pseudomonas aeruginosa and so on.24.0% was Gram positive bacteria,including staphylococcus aureus accounted for 10.5%,and the epidermis staphylococcus,hemolysis staphylococcus and so on.Proportion of fungi was 11.0%.According to the results of microbiological data,the effective anti-microbial treatments were administered.After treatments for 10 days, the VAP infection parameters(including temperature,the white blood cell count and the number of strains)were grad-ually back to normal.Conclusion The main VAP pathogenic bacteria are Gram negative bacteria,patients commonly are infected by two or more bacterias,which lead to the multiple infection,the treatment should be fully based on mi-crobiology and clinical monitoring data,and the formulating personalization antibacterial treatment.