Objective To study the inhibitory effects of siRNA targeting survivin on the expression of survivin,as well as the apoptosis,proliferation and invasion of a human melanoma cell line,M14.Methods Two siRNAs targeting survivin were designed,chemically synthesized,and used to construct the recombinant plasmids,pRAT-H1.1/neo-survivin-siRNA1 and pRNAT-H1.1/neo-survivin-siRNA2.Then,recombinant plasmids were transfected into M14 cells mediated by Lipofectamine 2000 reagent.Those cells untransfected or transfected with empty vector served as the control.After culture over various periods of time.cells were collected for the detection of mRNA and protein expression of survivin with RT-PCR and Western blotting,respectively,and for the examination of apoptosis and proliferation of M14 cells by flow cytometry and MTT methods,respectively.Also,Transwell assay was performed to detect the invasive capability of M14 cells.Results A statistical decrease in the mRNA and protein expressions of survivin was observed along with an increase in apoptotic rate(x2=31.55,P＜0.01)in M14 cells transfectcd with siRNA-containing plasmid compared with untransfected and empty vector-transfected cells.As MTT assay indicated,on day 4 after the transfcorion,the proliferation of M14 cells was inhibited by(55.4±4.3)%,(34.5±4.3)%and(13.3±4.6)%,with pRNAT-H1.1/neo-survivin-siRNA1,pRNAT-H1.1/neo-survivin-siRNA2 and empty vector,respectively:there was a significant difference among the three groups(P＜0.05).Decreased invasive capability was noticed in M14 cells transfected with siRNA-containing plasmid compared with untransfected cells(all P＜0.05).Conclusions The plasmid containing siRNA against survivin can specifically inhibit the expression of sarvivin,proliferation and invasion of tumor cells,and induce cell apoptosis.The inhibition of survivin expression by siRNA may be a rational approach to the gene therapy for malignant melanoma.

Objective To discuss the effect of pericardial devascularization plus gastric fundus transaction in advance schis?tosomiasis patients with portal hypertension. Methods Thirty?six advanced schistosomiasis patients with portal hypertension treated with devascularization plus gastric fundus transaction(a portal hypertension group),as well as 10 patients treated with modified Sugiura operation(a modified Sugiura operation group)in the Third People’s Hospital of Yangxin County since 2006 were chosen as the observation objects,and the clinical effects of the two groups were observed and compared. Results The op?eration time,indwelling time of stomach tube,time to taking food after operation,drainage tube removal time of the portal hy?pertension group were all shorten than those of the modified Sugiura operation group(all P<0.05). The hospitalization expenses of the two groups were(25 466.00 ± 2 888.48)Yuan and(34 517.10 ± 4 948.39)Yuan respectively,and the difference was al?so statistically significant(P<0.05). The incidence rates of portal thrombosis of the portal hypertension group and modified Sug?iura operation group were 33.33%(12/36)and 40.00%(4/10),respectively,and the incidence rates of rehaemorrhagia of the two groups 12 months after the operation were 16.67%(6/36)and 10.00%(1/10),respectively,but the differences had no sta?tistically significance(both P>0.05). In addition,1 case with delayed gastric emptying and 1 case with stomal leak of esopha?gus happened in the modified Sugiura operation group,while no corresponding complications happened in the portal hyperten?sion group. Conclusions Pericardial devascularization plus gastric fundus is a relatively easy procedure which has a good short?term clinical effect,and therefore it is suitable for application in primary hospitals. However,its long?term effect still needs fur?ther observation.

Some new types of microemulsion using phospholipids as the main surfactant were prepared for electrokinetic chromatography and the quantitative structure-retention relationship of neutral solutes in these microemulsion electrokinetic chromatography (MEEKC) systems was studied by solvation parameters model.By using dynamic coating capillary, and with dimethyl sulfoxide (DMSO) and dodecyl benzene as the marker of electroosmotic flow and microemulsion droplets, a total of 17 kinds of stable microemulsions containing soybean phospholipids or other surfactants were prepared and the linear salvation energy relationship equations were developed for these MEEKC systems with 26 small neutral compounds.The coefficients of linear solvation energy relationship (LSER) equations were used to evaluate the similarity of two MEEKC systems.Results indicated that LSER characteristics of phospholipids-MEEKC systems were similar to those of other microemulsion systems.The volume and hydrogen bond basicity of solutes were mostly contributed to the retention in MEEKC.The different types and concentration of oil phase had no evident influence on the retention.

A 7?year?old girl presented with a plaque on the left side of the face for 4 years. Skin examination revealed a well?marginated circular firm plaque measuring 6 cm × 5 cm in size on the left side of the face with purulent crusts on the surface, which was nontender to palpation. Histopathologic study showed ulceration and necrosis in some areas in the epidermis, diffuse dermal infiltration of lymphocytes, histiocytes and epithelioid cells with the presence of many neutrophilic granulocytes in some regions. Both periodic acid?Schiff(PAS)staining and acid?fast staining were negative. After 25 days of tissue culture, scotochromogenic Mycobacterium colonies grew, and colony smears showed positive acid?fast staining. The sequences of 16S rRNA and hsp65 genes of the fungal isolate showed 100%and 99%homology with those of Mycobacterium szulgai, respectively. After the treatment with rifampicin, isoniazid and ethambutol, the patient′s condition was improved.

BACKGROUND: In recent years, there are many studies designed to explain the protective effect of lidocaine on brain, but few about the therapeutic effect on traumatic cerebral edema. The content of aquaporin-4(AQP-4) in brain tissue is the highest and it has been proved that AQP-4participants in the formation of cerebral edema induced by cerebral trauma, cerebral infarction, eerebrai tumor and other reasons.OBJECTIVE: To observe the effect of lidocaine on the expression of AQP-4 of experimental rats following brain injury and analyze the therapeutic effect of lidocaine on brain edema.DESIGN: A randomized and control animal experiment.SETTING: Department of Anesthesiology, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: The experiment was carried out in the Institute of Brain Disease, the Medical School Hospital of Qingdao University between January and July 2004. Totally 65 three-month-old healthy male Wistar rats,were enrolled in the experiment and randomly divided into 3 groups: normal control group (n=5), model group (n=30) and treatment group (n=30). Thirty rats in the model group and treatment group were respectively assigned into 6 subgroups according to 6 different time points: 1,4,6,12,24 and 48 hours following brain injury, with 5 rats at each time point.METHODS: Animal models of brain injury at right parietal lobe were created according to the method from Feeney et al. As for the rats of normal control group, they only underwent operation to be injured at the corresponding part. Rats recovered the access to food and water 2 to 8 hours after operation. For the rats in the treatment group, they were intraperitoneally injected of lidocaine at the 1st, 4th,6th, 12th, 24th, and 48th hours following injury, 5 rats at each time point. The initial dosage was 30 mg/kg, then 15 mg/kg was maintained . Administration was conducted every 6 hours in 3 days; For the rats in the model group, they were intraperitoneally injected of 30 mg/kg normal saline and rats in the normal control group were given no special treatments. Water content of brain was calculated 5 days following brain injury with dry and wet weight method:water content of brain=[brain mass (wet)-brain mass (dry)]/brain mass(wet)×100%. Expression of AQP-4 of brain tissue of rats was detected with immunohistochemical method and cytomorphological change of brain tissue was observed under optical microscope.MAIN OUTCOME MEASURES: ① Water content of brain of rats in each group. ② Expression of AQP-4 of brain tissue of rats in each group.③ Pathological results of brain tissue of rats in each group.RESULTS: No rats died accidentally or for other factors in the process of experiment, finally, all the 65 rats entered the stage of result analysis. ①Compared with model group, administration of lidocaine within 6 hours following brain injury could significantly decrease the water content of brain tissue [1 hour after brain injury: (81.09±0.29)%, (83.04±0.25)% ,P ＜ 0.05];[4 hours after brain injury: (81.34±0.35)%, (83.31±0.48)%,P ＜ 0.05] ;[6 hours after brain injury: (82.01±0.21)%, (83.25±0.37)% ,P ＜ 0.05]. Compared with model group, administration of lidocaine could significantly decrease the expression of AQP-4 [1 hour after brain injury:(0.19±0.02), (0.24±0.03),P ＜ 0.05]; [4 hours after brain injury: (0.21±0.05 ), (0.25±0.05) ,P ＜ 0.05]; [6 hours after brain injury: (0.21±0.03 ),(0.24±0.02) ,P ＜ 0.05]. There were no significant differences of AQP-4expression and water content of brain tissue when administration was conducted at the 12th, 24th and 48th hours following brain injury (P ＞ 0.05). ②Under the microscope, AQP-4 positive cells presented vacuolus, they mainly lay in the edema area of peripheral part of trauma, cortex of traumatic side and around the blood vessel as well as astrocyte of white substance, choroid plexus and ependymal layer. ③ Necrosis was found in most cells in the central area of trauma and apoptosis in most cells in the peripheral area. Compared with model group, necrotic and apoptotic cells were significantly less within 6 hours following trauma, but not at the 12th,24th and 48th hours following trauma in the treatment group.CONCLUSION: High dosage of lidocaine can decrease the expression of AQP-4 and lighten cerebral edema following brain injury, but administration should be given as early as possible.

Objective To summarize the experience in nursing children with pediatric advanced intraocular retinoblastoma treated with 23-gauge microinvasive vitrectomy. Methods Eleven infants (12 eyes) suffered from advanced intraocular (D or E) retinoblastoma and treated with 23-gauge microinvasive vitrectomy. Preoperative psychological nursing, postoperative complication nursing and follow-up nursing were performed. Results The retinal tumors was completely removed in all 12 eyes. After the follow-up of a half year, the conditions in 8 eyes were stable, and the tumors in 4 eyes recurred. Visual acuity in 4 eyes improved and that in 1 eye remained unchanged. Conclusions Microinvasive vitreous operation can effectively remove advanced intraocular retinoblastoma. Active efficient perioperative nursing plays an important role for the prevention of complications and promoting the recovery of affected eyes.

Objective To investigate the expression level of CD226 mRNA in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE) and explore the relation between the gene expression and disease activity, and the relation between the gene expression and Gly307Ser polymorphism of CD226 was also examined. Methods CD226 gene was measured with real-time polymerase chain reaction (qRT- PCR) in PBMCs. The expression levels of CD226 gene in PBMCs were compared between 90 SLE patients and 30 healthy individuals. One-way ANOVA and pearson correlation were used for statistical analysis. Results The expression level of CD226 in the PBMCs of SLE patients (6.8±1.1) was significantly decreased compared to healthy individuals (26.5±6.7) (P＜0.01), while there was no association between mRNA level and genotype (P＞0.05). No correlation between ESR, CRP, ANA, SLEDAI scores, C3 and the expression level of CD226 gene was discovered. Conclusion In Hubei Chinese Han population, CD226-Gly307Ser locus is associated with the development of SLE, while T allele does not impact the expression of CD226 gene, thus the role of CD226 gene in autoimmune diseases should be explored in the future.

Background and purpose: Recently, survivin gene is one of the hot spots in tumor study. The purpose of the present study was to observe the impact of RNAi targeting survivin gene on tumor growth, apoptosis and radiosensitivity in nude mice xenograft of human cervical carcinoma. Methods: HeLa-s2, HeLa-NC, HeLa-U6 neo and HeLa cells were inoculated respectively in flank subcutaneous tissue of 24 female nude mice so as to establish xenograft models of human cervical carcinoma randomly. The tumor growth status was observed. The tumor volume was regularly measured and the tumor weight was investigated used to observe the impact of RNAi targeting survivin gene on tumor growth. The expression of survivin protein and FⅧRAg in tumor tissues were examined by immunohistochemistry SP method MVD was calculated. Cell apoptusis in tumor tissues was observed by HE staining,apoptotic index(AI) was quantified by TUNEL method. To observe the impact of RNAi targeting survivin gene on tumor radiosensitivity after irradiated, the tumor growth delay and tumor weight were investigated and cell apoptosis were examined by TUNEL method. Results: We established 4 groups of xenograft models of human cervical carcinoma.The tumor volume of HeLa-s2 group was obviously less than that of HeLa group at every checkpoint. The tumor weight of HeLa-s2 group was obviously lower than that of HeLa group, and they were(0.369±0.043)g and (1.150±0.136)g respectively(P<0.05).The tumor growth inhibitive rate of HeLa-s2 group was 67.9%. The expression of survivin protein FⅧ RAg examined by immunohistochemistry in tumor tissues of HeLa-s2 group decreased sharply and MVD went down to 23.4±3.1. Apoptotic cells increased in tumor tissues of HeLa-s2 group examined by HE staining and TUNEL method, AI was (22.74±1.4)%. The tumor volume of HeLa-s2 group was obviously less than that of HeLa group at every checkpoint after irradiation and the tumor weight of HeLa-s2 group was obviously lower than that of HeLa group, they were:(0.41±0.06)g and(1.38±0.29)g respectively(P<0.05). Cell apoptosis increased in tumor tissues of HeLa-s2 group.Compared with HeLa group, AI of HeLa-s2 group rose up notably, they were(30.06±0.98)% and (4.17±0.64)%(P<0.05).Conclusion: RNAi for survivin gone can reduce MVD in xenograft by inhibiting survivin protein expression, thus inhibit tumor growth and induce cell apoptosis, Survivin gene RNAi can also enhance tumor radiosensitivity.

Objective To analyze the epidemiological,clinical and histopathological characteristics of cutaneous leishmaniasis.Methods This study included six patients with cutaneous leishmaniasis diagnosed in the Institute of Dermatology,Chinese Academy of Medical Sciences and Peking Union Medical College,over the past 10 years.The epidemiological features as well as clinical and histopathologic presentations of these patients were analyzed retrospectively.Results All of the six patients were male.The mean age at onset of skin eruptions was 47.67 (range:37-67) years,and the mean duration of disease was 10 (range:6-18) months.Clinical presentations included erythema,nodules and ulcers in the face and limbs.Skin biopsy revealed infection-associated granulomatous inflammation with many amastigotes (basophilic bodies) in the cytoplasm of histiocytes,which were highlighted with Giemsa stain.All the patients had a history of working at or travel to epidemic areas.Conclusion The diagnosis of cutaneous leishmaniasis mainly depends on epidemiological data,clinical manifestation and histopathologic findings.

Objective To study the effects of 5-aza-dc on the expression of IGFBP7 in and proliferation of melanoma cell lines A375 and M14.Methods Reverse transcription-PCR and immunocytochemistry were performed to detect the mRNA and protein expression of IGFBP7 in A375 cells and M14 cells after treatment with 5-aza-dc of 10μmol/L for 48 hours,and MTT assay to measure the proliferation of both cell lines treated with 4 different concentrations (2.5,5,10,20μmol/L) of 5-aza-dc for various durations.Results The treatment with 5-aza-dc restored IGFBP7 expression at both mRNA and protein levels.The four concentrations of 5-aza-dc inhibited the proliferation of A375 and M14 cells in a dose-dependent (F=561.12,271.43,respectively,both P<0.01) and time-dependent (F=141.35,549.33,respectively,both P<0.01) manner.Conclusions DNA methylation may be involved in the modulation of aberrant IGFBP7 gene expression in melanoma,and 5-aza-dc could inhibit the proliferation of A375 and M14 cells.