Carbon-Carbon Ligases ; deficiency ; Child ; Gas Chromatography-Mass Spectrometry ; Humans ; Male ; Prognosis

OBJECTIVETo investigate the apoptosis-inducing effect of DMF on the human liver cells (HL-7702) in vitro.

METHODSLiver cells were exposed to different concentrations of DMF (0, 50, 100, 150, 200 mmol/L) for 12 hours. Apoptotic rate, the expression of Bax, Bcl-2 and Caspase-3 in liver cells were measured by FCM and western blotting respectively.

RESULTSThe increase in apoptotic rate of hepatocytes in concentration-manner was shown after DMF treatment for 12 h. After treatment the expression of Bcl-2 was decreased steadily and lower than the control group (P < 0.01), the expression of Bax showed no significant difference among the groups of different dosage by one-factor analysis of variance (P > 0.05), as the increase of the dosage of DMF. The ratio of Bcl-2/Bax dropped with the dosage of DMF increasing, and the ratio in 200 mmol/L of DMF was significantly lower than that of the control (P < 0.01). The new lands of procaspase-3 in 150, 200 mmol/L were observed, which demonstrated that there was active caspase-3.

CONCLUSIONDMF can induce apoptosis of cultured adult normal hepatocytes in vitro, and the mechanism might be related to the decrease of Bcl-2/Bax and the cleavage of Caspase-3.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cells, Cultured ; Dimethylformamide ; pharmacology ; Hepatocytes ; drug effects ; metabolism ; pathology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

AIM: To study the effects of tetrandrine(Tet) and fructose-1,6-diphosphate(FDP) on the elevated intrasynaptosomal [Ca 2+ ] i induced by excitatory amino acids (EAA). METHODS: A rapid method for preparing synaptosomes was used, and intrasynaptosomal free calcium ([Ca 2+ ] i) was measured by using the fluorescent indicator quin-2. RESULTS: L-glutamate (Glu, 100 ?mol/L), aspartate (Asp, 100 ?mol?L -1 ), N-methy1-D-aspartate (100 ?mol/L) and Glu (50 ?mol/L) plus Asp (50 ?mol/L) all elevated intrasynaptosomal [Ca 2+ ] i in a dose-dependent manner. Pretreatment with Tet (10,30,60 ?mol/L) , FDP (15, 30, 75, 150 ?mol/L), MK-801 (10, 20 ?mol/L) and Tet (15, 30 ?mol/L) plus FDP(15, 30 ?mol/L) all attenuated the increase in intrasynaptosomal [Ca 2+ ] i induced by EAAs mentioned as above in a dose-dependent manner, and the effect of Tet plus FDP was most significant. CONCLUSION: Both Tet and FDP inhibited a rise in intrasynaptosomal [Ca 2+ ] i induced by EAAs, which may be one of mechanisms that Tet and FDP pretect cerebral tissues against ischemia injury.

Background Cell immunologic therapy for retinoblastoma(RB)is becoming a hot research topic.Cancer-testis antigen is a human immunogenic protein and is used to treat some tumors.However,its effect on RB has not been investigated.Objective The present study was to discuss the antigen specific anti-tumor effect of cytotoxic T lymphocytes(CTL)induced by the cancer-testis antigen,NY-ESO-1-sensitized dendritic cells(DCs),on human RB.Methods PCR was performed to amplify target gene fragments from the NY-ESO-1 plasmids,and then the target gene fragments were digested with the restriction enzymes SalI and EcoRI.Harvested fragments were inserted into the pDC316 plasmid to construct the recombinant plasmid pDC316/NY-ESO-1.The expression of NY-ESO-1 protein in human RB cells strain,HXO-RB44,was detected by immunofluorescence and Western blot.Monocytes were isolated from 60 ml of peripheral blood from a healthy donor using Ficoll density-gradient centrifugation with a cell density of 1 × 107/ml.DCs isolated from blood were stimulated with recombinant human granulocyte-macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin-4(rhIL-4).The recombinant plasmid pDC316/NY-ESO-1 was transfected into DCs and the DCs were co-cultured with T lymphocytes.The resultant CTL were used as effector cells.The growth of the CTL was detected by MTT assay.The CTL were then added into the growth medium used for culturing HXO-RB44 cells and the vitality of the HXO-RB44 cells was assayed by MTT assay.Results The sequence of the cloned DNA fragment of the recombinant plasmid pDC316/NY-ESO-1 was conforms with the sequence of the NY-ESO-1 gene.The expression of the NY-ESO-1 protein in HXO-RB44 cells was tested by immunofluorescence and Western blot.DCs were successfully induced with rhIL-4 and rhGM-CSF from PBMC.The recombinant expression plasmid pDC316/NY-ESO-1 was successfully transferred into DCs.These DCs had high expression of surface molecules such as HLA-DR(42.1％),CD80(54.2％),CD83(39.7％)and CD86 (94.8％).The CTL that was induced by DCs-sensitized with NY-ESO-1 specifically killed HXO-RB44 cells.CTL induced by the sensitized DCs had a stronger cytotoxic effect against HXO-RB44 cells compared with un-sensitized DCs and CTL un-induced with DCs,as shown by MTT asssay(P＜0.05).The anti-tumor activity was highest when the ratio of effector to target was 75∶1(P＜0.05).Conclusions DCs transfected by the recombinant plasmid pDC316/NY-ESO-1 can induce the proliferation of allogenic CTLs,which showed a specific anti-tumor effect against HXO-RB44 cells.These results present a new type of immunotherapy for the treatment of RB.

AIM:To evaluate the efficiency of vitrectomy combined with inferior scleral buckling for a special kind of retinal detachment.
METHODS: Nineteen eyes of special retinal detachment had following features: 1 ) the course was more than 6mo;2) there were at least one hole located in the 5 : 00 - 7 : 00 of the marginal retina; 3 ) the detachment range of retina more than 270 degrees and retinal proliferation in retinal detachment. The 19 eyes who received vitrectomy combined with inferior scleral buckling, retinal anatomic reattachment and visual function recovery was observed.
RESULTS: Among these 19 eyes, all eyes retinal anatomic reattachment and best corrected visual acuity ( BCVA ) was improved in various degrees. The BCVA was 0. 01-0. 1 in 5 eyes, 0. 12-0. 3 in 9 eyes, and ≥0. 4 in 5 eyes.
CONCLUSION: It is an effective method to vitrectomy combined with inferior scleral buckling for the special kind of retinal detachment which has following features:1) the course was more than 6mo;2) there were at least one hole located in the 5:00-7:00 of the marginal retina;3 ) the detachment range of retina more than 270 degrees and retinal proliferation of cable in retinal detachment.

This article reported the treatment of 3 cases of frequent recurrent intractable nephrotic syndrome by XIE Chang-sen. On basis of the therapeutic method of invigorating the spleen and kidney, other methods should be used according to individual difference and special clinic signs and symptoms, such as other invigorating qi and consolidation of superficies, nourishing blood and dispersing stagnated liver qi, warming kidney qi to invigorate yang, replenishing essence and dispersing turbid and so on. And partner treatment is the key for cure.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Graft vs Host Disease ; therapy ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunosuppressive Agents ; therapeutic use ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Salvage Therapy ; methods ; Treatment Outcome ; Young Adult

Anticonvulsants ; adverse effects ; Asian Continental Ancestry Group ; genetics ; Carbamazepine ; adverse effects ; China ; epidemiology ; Drug Eruptions ; epidemiology ; ethnology ; genetics ; Genetic Predisposition to Disease ; genetics ; HLA-B Antigens ; genetics ; HLA-B15 Antigen ; genetics ; Humans ; Risk Factors ; Severity of Illness Index ; Skin ; pathology ; Stevens-Johnson Syndrome ; epidemiology ; etiology ; genetics

The purification of kallikrein using CTAB/hexanol/octane reverse micelles extraction has been studied and optimized,under various aqueous pH values, ionic strength and species, CTAB concentration and co-surfactant concentration. The result shows that the extraction efficiency approaches 100%, and the activity recovery is more than 80%, the commercial enzyme specific activity is increased by 1.97 times and the crude enzyme activity is increased by 7.15 times, which from 31U/mg to 219U/mg,under the conditions of[CTAB]=0.02 mol/L, hexanol/octane (V/V)=1∶5, pH=9.0 and[KBr]=0.1 mol/L in forward extraction, pH=7.0 ,[KBr]=1.5 mol/L and 15% ethanol(V/V) in backward extraction. The result of purified kallikrein is examined by the SDS-PAGE analysis. Reverse micelles extraction is a potential technique for the application in the downstream biotechnological processes.