Objective To observe the effects of hemin on the expression of hemeoxygenase(HO) on the human umbilical vein endothelial cells.Methods The hemin and zinc protoporphyrin-Ⅸ were added in the cultured endothelial cells to induce and inhibit the expression of HO-1,respectively.The expressions of HO-1 and HO-2 mRNA were detected using reverse transcription polymerase chain(RT-PCR) and the relative amount of CO released into the media was quantitated as carboxyhemoglobin(COHb) by enzyme linked immunosorbent assay(ELISA).The proliferation of endothelial cells was detected by methyl thiazolyl tetrazolium test(MTT).Results The expression of HO protein was mainly detected in the cytoplasm of endothelial cell.The expression of HO-1 mRNA in endothelial cell and the amount of COHb in the media increased significantly with the treatment of hemin.Conclusion The expression of HO-1 and the increase of endogenous CO are the molecular bases of the proliferation of endothelial cells,and the HO/CO system may play an important role in the development of cardiovascular diseases.

Objective:To find out the role of JNK/SAPK signaling-transduction pathway in the effect of elemene against hepatocarcinoma, offering the clue to ilustrate the molecular mechanisms of antitumor effects of elemene. Methods: The detection of the distribution of elemene in Hca-F cells was detected by gas chomatography and apoptotic changes in elemene treated. SMMC7721 cells were examined by TEM. After elemene treatment, the activation of JNK/SAPK in HepG2 cellls and the DAXX gene expression in SMMC7721 cells were detected by Western blotting and RT-PCR respectively. Results: Gas chomatography showed that elemene was detected at 8. 42 minute. The SMMMC7721 cells treated by elemene for 3 hours began to show typical apoptotic changes . The JNK/SAPK activity in HepG2 cell treated with heat shock was the highest of all groups and the group treated with elemene was the next and the control group is the lowest one. There was no DAXX gene expression in SMMC7721 cells treated with elemene. Conclusion: Elemene can diffuse into cells. Tumor cell apoptosis treated with elemene may be induced by JNK/SAPK activating and DAXX signal pathway may not play key role in JNK/SAPK activation induced by elemene.

The expression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) in the placenta of the patients with pregnancy induced hypertension (PIH) was detected and its role in the pathogenesis of PIH was studied. The pathological changes in placental vessels were observed by HE staining. NO2-/NO3-, the stable metabolic end products of NO, was measured with nitrate reductase. The eNOS activity in placental tissues was assayed by spectrophotometry. Western blot analysis was applied to detect NOSTRIN expression. The incidence of thickening and fibronoid necrosis of placental vessels was significantly higher in women with PIH than in the normal group (P < 0.01). The levels of placental NO2-/NO3- in PIH patients (27.53 +/- 7.48 micromol/mg) were significantly lower than in normal group (54.27 +/- 9.53 micromol/mg, P < 0.01). The activity of eNOS was significantly decreased in PIH group (12.826 +/- 3.61 U/mg) as compared with that in normal group (21.72 +/- 3.83 U/mg, P < 0.01). Western blot analysis revealed that both groups expressed 58 kD NOSTRIN, but the protein level was significantly higher in women with PIH than in the normal group (P < 0.01). A significant negative correlation existed between the expression of NOSTRIN protein and the activity of eNOS in placental tissue of women with PIH (r = -0.57, P < 0.01). It was concluded that the level of NOSTRIN expression in placenta of women with PIH was increased, which may play an important role in the pathogenesis of PIH.

Objective: To investigate the effect of moxibustion on telomerase activity and genes expression in tissues of senescent rats. Methods: Subacute aging rats model were established by injection with D-gal solution. Points Shenshu (BL 23) were treated with moxibustion in treatment group, contrasting with a model group and a normal group. Enzyme-Linked Immunosorbent Assays(ELISA) was used for the level oftelomerase activity in liver tissues, and In Situ Hybridization(ISH) was used for the condition of expression of telomerase genes in liver tissues. Results: The level of telomerase activity in the aging model group was obviously lower than that in normal control group (P＜0.01), the level in moxibustion group was obviously higher than that in model group(P＜0.05). In comparison with normal rats, the positive-expressed areas and photodensity of telomerase genes in aging model group were all significantly lower (P＜0.01,P＜0.01), and the positive-expressed areas in moxibustion group was significantly higher than that in model group (P＜0.01). Conclusions: Moxibustion could regulate telomerase activity of senile rats, hence delaying aging.

Objective To investigate the effect of bibliotherapy in soothing the postoperative pain in children and relieving the perioperative anxiety of children and their care givers. Methods Hospitalized children and their care givers from August 2007 to March 2009 were studied. 153 cases from August 2007 to May 2008 were assigned to the control group and the other 153 cases to the intervention group. Routine surgical nursing were applicated in the control group by introduction of perioperative nursing procedures.Bibliotherapy were applicated in the intervention group on the basis of the control group-using "bibliotherapy materials for hospitalized children" which was designed by ourselves and correspond with the theory of bibliotherapy to interfere in the 153 cases in the intervention group. The variance of preoperative anxiety of care givers and perceptions of postoperative pain of children between the two groups were compared with scales of mYPAS, STAI and FLACC and Wong- Baker Facial Scale. Results The scores of mYPAS in children of the intervention group and the control group was (35.875+4.441)and(46.796+8.606 )respectively and the variance was significant. The scores of STAI in care givers of the two groups was(38.125+4.371 )and (49.901 +7.420) respectively and revealed significant variance. The scores of Wong-Baker and FLACC in children of both groups 1 hour after operation were compared and revealed no statistical significance. The scores of Wong-Baker and FLACC in children at 6 hours and 24 hours postoperative were compared subjectively and objectively and revealed statistic significance. Conclusions Bibliotherapy can ameliorate the anxiety level of both children and their care givers, relieve the perception of postoperative pain in children and improve their comforts. Bibliotherapy thus conduces to the recovery of postoperative children.

To investigate the expressions of placental growth factor (PLGF) in placenta with hypertensive disorders of pregnancy (HDP), 45 women with HDP and 20 normally pregnant women were studied. Among 45 women with HDP, there were 23 cases of severe preeclampsia and one case of eclampsia. The location and level of PLGF proteins was determined by immunohistochemistry and Western blot. The expression of PLGF mRNA in placenta was assessed by reverse transcriptional-polymerase chain reaction (RT-PCR). The results showed that: (1) The distribution of PLGF in placenta with HDP was similar to normal one, which was mainly in the cytoplasm of villous syncytiotrophoblast and villous stroma; (2) The expression of PLGF protein was significantly decreased in placentas with mild and severe preeclampsia compared to the normal ones (0.3 +/- 0.4 vs 0.6 +/- 0.4, 0.2 +/- 0.5 vs 0.6 +/- 0.4, P < 0.01). There were no differences between the gestational hypertension placenta and normal one (0.5 +/- 0.6 vs 0.6 +/- 0.4, P > 0.05); (3) The transcription levels of the PLGF mRNA in placentas with preeclampsia were significantly lower than in normal groups (3.33 +/- 0.39 vs 4.87 +/- 0.60, 1.97 +/- 0.29 vs 4.87 +/- 0.60, P < 0.01), and no differences were found between the gestational hypertension placenta and normal groups. These findings suggest that the abnormal expression of PLGF in placentas is related to the pathogenesis of HDP.
Placenta/*metabolism ; Pre-Eclampsia/*metabolism ; Pregnancy/*metabolism ; Pregnancy Proteins/*biosynthesis ; Pregnancy Proteins/genetics

Objective Radiofrequency ablation (RFA) and endoscopic breast conserving surgery (EBCS) has been developed as a potential means for treating breast tumors with minimal disruption to adjacent soft tissues.The purpose of this study was to evaluate the cosmetic outcomes of RFA assisted by EBCS on patients with early breast carcinoma (T1 -2 N0-1 M0).Methods Twenty-three patients with biopsy proven invasive breast carcinoma ( ≤3 cm in diameter),were treated with ultrasound (US)guided percutaneous-RFA for the local tumors followed by sentinel lymph node biopsy and endoscopic auxiliary node dissection and lumpectomy.The reactions to breast cosmetics were observed and compared to the conventional approaches using the breast asymmetry score for EBCS and a self -assessment questionnaire targeting objective and subjective data about treatment related breast change.Results Twenty-three patients were successfully treated with RFA.Complete coagulation necrosis of the tumor was visualized at US and mammography in 22 of 23 patients (95.6 ％).22 patients being per formed RFA and EBCS presented little scars and the breasts preserved more cosmetically acceptable than the reported results of traditional protocols.Self-assessment acceptable rate was 100 ％,aesthetic outcome as good or excellent rated in 95.5 ％(21/22).Within the follow-up of above 36 months,there were no local recurrence,metastasis,wound infection or necrosis,nerve damage,haematoma,as well as upper limb swelling or lymphoedema,no reduced upper limb mobility and tumor seeding.Conclusions RFA with endoscopic breast conservation therapy for the treatment of early breast carcinoma is more effective and cosmetically acceptable than the traditional approaches.

Objective To develop and evaluate the allele-specific real-time PCR(ASPCR) assay for the detection of minor HIV-1 variants. Methods We developed and evaluated the ASPCR assay, using the K103N mutation site as a model system. We constructed plasmids as standards and designed specific and non-specific primers to discriminate the wild-type and mutant plasmids in the real-time PCR using SYBR green as fluorescence reporter. And then we evaluated the sensitivity, accuracy, reproducibility of ASPCR assay and detected the control samples. Results The specific primer can discriminate the wild-type and mutant plasmids including resistant mutation successfully. The sensitivity of ASPCR assay can achieve less than 0.01% and the accuracy of this method is down to 0.1%. The Intra-assay coefficient of variation is less than 0.7 and the Inter-assay coefficient of variation is less than 1.6. Conclusion ASPCR is a sensitive, accurate and rapid method to detect the minor HIV-1 variants which have resistant mutations and it can be used widely in HIV research. ASPCR also can provide earlier and more resistant information to the clinical therapy.

Objective To isolate stable passage primary HIV-1 drug resistant strains and observe replication dynamics of the drug resistant isolates and evolvement tendency of the drug resistant mutations in vitro.Methods Peripheral blood mononuclear cells(PBMCs)from 15 AIDS patients receiving highly active antiretroviral therapy(HAART)were collected,and the primary HIV-1 stains were separated utilizing co-cultivated with PBMCs from normal people.HIV-1 pol genes from those strains were obtained by RT-PCR and sequenced.The drug resistant mutations were analyzed in the Stanford HIV Drug Resistance Database.Results Eight strong positive strains were isolated from 15 AIDS patients with viral loads higher than 1000 copies/ml,and two of them were drug resistant.Drug resistant mutations of the two strains were respectively K103N/K238T and M184V/K103N/Y181C/H221Y which show high-level resistance to NVP and 3TC/NVP,respectively.K103N,M184V,Y181C and H221Y exist stably in the environment without drug pressure,however,RT K238T reverted to K238.Conclusion Two drug resistant strains were successfully isolated in vitro without drug pressure.Strains with K103N shows superior fitness and can exist steadily.Strains with M184V and K103N/Y181C/H221Y can also replicate stably in vitro without drug pressure.NNRTI mutation K238T reproduces astatically,which suggests that RT 238 codon might revert gradually to wild genotype.

Objective To evaluate the antiretroviral therapy(ART),analyze the prevalence of resistance in rural areas,Henan,and explore the presence of minor resistant variants in pre-ART.Methods One hundred and forty-nine AIDS patients initiating ART were recruited and investigated at intervals of 6 months. Method of In-house developed by our laboratory for genotypjc resistance test was to analyze the occurrence of resistance among the failure of ART,and the allele-specific real.time PCR(ASPCR)was used to detect the minor resistant variants at the baseline samples once the resistance occurred.Results Vimlload significantly decreased among the patients who received ART(t=275,P=0.0001),but the absolute counts of CD4+T lymphocytes had no significant change(t=1.765 168,P=0.0852).Rate of resistance among the patients of treatment failure was 4.88%.The result of ASPCR in the survey of baseline showed that the minor resistant variants of M184V were detected in 7 patients and mutation K103N presented in 5 patients.Conclusion The primary drug-resistant straias in the untreated patients were found in Henan,and they might develop the dominant resistance strains and bring about the failure of ART.