The whole length cDNA of human interferon-?(IFN-?) gene including signal peptide was constructed to retroviral vector pLXSN and then packaged with PA317. A Human Hepetoma Cell Line(HHCL) and a human gastric cancer cell line (MKN-45) were infected by using supernatant containing retroviral vector with IFN-?. the gene-modified tumor cell. were isolated by G418 selection. Some changes were found in cell cycle, expression of HLA class Ⅰ and classⅡ, and tumorigenecity. Elevated immunogenecity and abrogated rumerigenecity suggest a means for generating gene-modified tumor cell vaccine. for the treatment of cancer.

Retroviral vector was used to introduce the interleukin - 2 (IL -2) gene into H22 cells, a BALB/c mouse hepatoma cell line. The IL-2 gene and marker gene (NeoR) were assessed by using PCR and RT-PCR methods. FACS analysis demonstrated that the cells with low DNA content in IL-2 gene modified H22 cells were more than these in control H22 cells. Electron microscopic morphological structure showed that some cells in IL-2 gene modified H22 went into apoptotic cell death. The study demonstrated that apoptotic cell death of H22 - IL-2 cells was induced by the IL-2 gene modification. It may be one of mechanisms of decreased tumorigenicity of IL - 2 gene modified tumor cells.

We compared the ability of tumorigenicity of TNF-? gene-modified and unmodified H22 tumor cells, and therapeutic functions of the irradiated cells. Results indicated that H22-TNF-? cells were less tumorigenic as compared to H22 and H22-LXSN cells. In case of treatment groups, injected with irradiated tumor vaccine in 3 or 6 days after inoculation of H22 cells, the tumor growth was suppressed.

Retroviral vector pLXSN was employed to introduce human r-Interferon (IFN-?) gene into four human hepa-tocellular carcinoma cell lines (HCC). The G418-resistant colonies were isolated and cloned. PCR and RT-PCR analysis indicated that the integration and expression of IFN-? gene was shown only in the transduced cells. Using a bioassay method, we found that all genetically modified HCC cells can secrete varied amount of IFN-?. The results of flow cytome-try showed that the cell surface expression of HLA class I molecules significandy increased following transduction. Moreover , we firsdy indicated that the increase in the expression of one specific HLA class I antigen, HLA-A2, was almost in the same magnitude as that of the total HLA class I molecules after transduction with IFN-r gene.

Objective To discuss the impact of hyperthyroidism on pregnancy outcome.Methods Clinical data of 80 pregnancy patients with hyperthyroidism(hyperthyroidism group)were retrospectively analyzed,in accordance with the norms of treatment during pregnancy is divided into treatment group(61 cases)and untreated group(19 cases);At the same time,in accordance with the ratio of 1:2 160 cases of healthy pregnant women were randomly selected as control group.childbirth,the maternal and perinatal outcome in three groups were compared.Results Graves disease group the incidence of pregnancy-induced hypertension 10.0%,3.8% incidence of heart failure,cesarean section rate of 61.4%,7.6% incidence of prematurity,asphyxia incidence of 7.6%,a low incidence of 3.8%,the mortality rate 3.8%,and 1.3% of the control group,41.3%,1.3%,0.6%,0,0,the differences were statistically significant(P＜0.05),the incidence of neonatal abnormalities in two groups is not different(P＞0.05);the treatment group of diseases of pregnancy-induced hypertension,heart failure,premature birth,asphyxia,such as the incidence were 3.3%,3.3%,3.3%,0,0 mortality,with non-governing group of 31.6%,15.8%,21.1%,15.8%,10.5%.The differences were statistically significant(P＜0.05),and a low incidence of the two groups,the cesarean section rate,incidence of deformities such as no difference.Conclusion Hyperthyroidism can increase the incidence of pregnancy complications,and standardize the treatment can improve pregnancy outcome.

AIM:To study the protective effects of penehyclidine hydrochloride(PHC) on transient forebrain ischemia reperfusion injury in gerbils.METHODS:The model of transient forebrain ischemia reperfusion was established in gerbils by bilateral carotid artery clamping.The effects of PHC on neurological function scores and the morphous of hippocampal pyramidal neuron of gerbils were observed after receiving transient forebrain ischemia reperfusion.SOD activities and contents of MDA in the hippocampus and cortex of gerbils were measured.RESULTS:In the groups of PHC(0.08),(0.24)(mg?kg~(-1)) and atropine,the stroke index was decreased,compared with the ischemia-reperfusion group after the gerbils received transient forebrain ischemia reperfusion for six hours.PHC could reduce the degree of injury in hippocampal pyramidal neuron after ischemia reperfusion for three days.CONCLUSION: PHC has protective effects on transient forebrain ischemia reperfusion injury in gerbils.

Objective To investigate the effects of the expressions of thymidine kinase 1 (TK1) and Ki67 alone or their combination on the recurrence and metastasis of breast cancer.Methods Sixty-five samples were selected from the Second Affiliated Hospital of Guangzhou Medical University from March 2005 to June 2007,which were resected by surgical operation and confirmed as breast carcinoma by pathology.They were individed into two groups including 37 cases with recurrence or metastasis in 5 years (group A),28 cases without recurrence or metastasis in 5 years (group B).The expressions of TK1 and Ki67 in the two groups were detected by immunohistochemical staining assay.Then,Kaplan-Meier assay was used to describe survival curve.Results The positive expression rate of TK1 in group A was 91.8％,which was dramatically higher than that in group B 67.8％ (x2 =6.116,P =0.013).The positive expression rates of Ki67 in group A and B were 78.4％ and 42.9％ (x2 =8.635,P =0.003).The positive expression rates of TK1 combined with Ki67 in group A and B were 67.6％ and 39.3％ (x2 =5.159,P =0.023).Moreover,disease free survival of patients with positive expression of TK1 combined with Ki67 decreased significantly,compared with patients with positive expression of TK1 or Ki67 alone (x2 =6.137,P =0.046).Conclusion Positive expression of TK1 combined with Ki67 is the high risk factor of the reccurence or metastasis of breast carcinoma,and indicates poorer prognosis compared with positive expression alone.

Objective: To explore the effect and underlying mechanism of puerarin on cell proliferation and ossification of fibroblasts in ankylosing spondylitis (AS). Methods: Fibroblasts were isolated from the hip capsule tissue of AS patients or femoral neck fractures. The cells were treated with puerarin at different concentrations (25, 50, 100, 150, and 200 mg/L). Cell proliferation was measured by CCK-8 method. Protein expressions of IL-6, TNF-α, OCN and OPN were measured by Western blot. ALP activity was detected and the mRNA expression of Col I was evaluated by qRT-PCR. Results: Puerarin of 150 and 200 mg/L significantly inhibited the proliferation of normal fibroblasts, while puerarin of 100, 150 and 200 mg/L significantly inhibited the proliferation of AS fibroblasts. Puerarin significantly inhibited protein expressions of pro-inflammatory cytokines IL-6 and TNF-α, decreased ALP activity, inhibited mRNA expression ofCol, and attenuated protein expressions of OCN and OPN. Conclusion: Puerarin inhibits cell proliferation and fibroblast ossification of normal and AS fibroblasts.

Objective To investigate the protective effects of Trimetazidine(TMZ)on the ischemia reperfusion injury (IRI)of fatty liver in autotransplantation model. Methods Fatty liver model was established by feeding high fat diet. Male Wistar rats (n=30) were randomized into three groups;Sham group, TMZ group and Model group. Liver was autotransplanted in both TMZ group and Model group. Serum levels of ALT, SOD, MDA, Bcl-2 and activated Caspase-3 were assessed 6 hours after the operation. The pathological performances of liver were also determined. Results Compared with the Model group, serum levels of ALT,AST, MDA and SOD levels decreased significantly in the TMZ group(P<0.05). Serum level of Bcl-2 was higher while level of activated Caspase-3 was lower in TMZ group than those in Model group(P<0.05). Histo?logical assay and TUNEL staining showed reduced hepatocyte swelling and narrowed sinusoid as well as decreased hepatic apoptosis in TMZ group compared with Model group. Conclusion TMZ can reduce oxidative stress, promote the expression of Bcl-2 and inhibit the activation of Caspase-3, which all contribute to its protective effect on fatty liver with ischemia-re?perfusion injury.

Objective To explore the disruption of circadian rhythm induced by the alteration of il?lumination leads to cognition impairement and mood dysreguation in mice. Methods 36 male C57BL/6 mice were divided into constant light group( CL) ,normal light group( N) and constant darkness group( CD) . Open field test was applied for the comparison of locomotor activity,tail suspension test and forced swimming test were conducted for assessment of mood state,and elevated plus?maze and Morris water maze were con?ducted for assessment of cognitive function. Results ( 1) Different circadian rhythms did not change the lo?comotor activity among three groups (CL:(200 160.00±955.28)cm,N:(208 148.00±578.11)cm,CD:(179 128.00±1 185.80)cm, P>0.05). (2) Compared with the control group,CL and CD mice showed significantly decreased immobility time in both TST (CD:(40.16±3.82)s,N:(18.83±2.27)s,CL:(46.00±2.80)s, P<0.01) and FST(CD:(181.33±9.03)s,N:(118.83±7.68)s,CL:(151.83±3.06)s, P<0.05). (3) Compared with the control group,CL and CD mice spent less time (CD:(21.76±6.88)s,N:(80.67±11.19)s,CL:(12.50±5.23)s, P<0.05) and made fewer entries (CD:3.33±0.49,N:6.83±0.91,CL:2.00±0.77, P<0.05) into open arms in elevated plus?maze, and exhibited less crossings in target quarter in Morris water maze (CL:2.67±0.76,N:5.00±0.26,CD:2.83±0.40, P<0.05). Conclusion Chronic constant light or darkness leads to negative impacts on mood and cognition in mice.