Objective To construct the specific small hairpin RNA(shRNA)plasmids for Formin-like 2(FMNL2)gene,and to observe its effect on proliferation of SW620.Methods Plasmids FMNL21 shRNA and plasmids FMNL22 shRNA complimentary to the sequence responsible for the function domain of human FMNL2 were constructed and transfected into SW620,respectively.The inhibition rates of FMNL2 expression were detected by re-al-time PCR after transfection with PGenesil-FMNL21 and PGenesil-FMNL22,respectively.The expression of FMNL2 was detected by real-time PCR at 12,24,48,72 and 96h after transfection.At 48 h and 72 h after transfection,the proliferation of SW620 was detected using MTT assay.Resuits The transfection rate in SW620 was(60.8±2.8)%for PGenesil-FMNL21,(58.7±2.9)%for PGenesil-FMNL22,and(62.6±1.7)%for PGenesil-HK(P＞0.05).The inhibition rate of FMNL2 expression after transfection with PGenesil-FMNL21 was 77.1%,which was the highest.The inhibition rates of FMNL2 expression were 13.5%,57.3%,80.3%,62.4%,and 33.2%at 12,24,48,72and 96h after transfection respectively.The proliferat capability of SW620 was obviously lower than that of control cells(P＜0.05).Conclusion Both plasmids FMNL21 shRNA and plasmids FMNL22 shRNA can inhibit the expression of FMNL2,of which Pgenesil-FMNL21 execs the greatest role.FMNL2 could be related to the proliferation of SW620.

Objective To analyze the relationship between the expression of formlin-like 2(FMNL2)and colorectal carcinoma metastasis through the detection of the expression of FMNL2 in colorectal carcinoma tissue and paracancerous tissue.Methods ①Immunohistochemistry was used to detect the expression of FMNL2 in 175 cases of paraffin wax specimens(including 30 cases of normal mucosa,25 cases of adenoma,75 cases of colorectal carcinoma and 45 cases of metastatic tumors).②Real time-PCR was used to detect FMNL2 mRNA expression in 32 cases of fresh specimens of carcinoma and 32 cases of paracancerous tissue.Results The positive detection rate of FMNL2 was higher in colorectal carcinoma tissue than in paracancerous tissue(P=0.003),higher in lymphatic metastasis than in primary carcinoma tissue(P=0.037),and was aslo higher in metastatic carcinoma tissue than in non metastatic carcinoma tissue(P=0.011).The expression of FMNL2 mRNA was not related to the infiltration and differentiation of carcinoma tissue but its concentration in metastatic carcinoma tissue was 2.5 times higher than in non metastatic carcinoma tissue.Conclusion FMNL2 gene may play an important role in the metastasis of colorectal carcinoma.

[Objective]To analyze reason of congenital infantile coxa vara to be misdiagnosed as infantile dislocation of hip joint.[Method]There were 6 patients(6 hip join)congenital infantile coxa vara being misdiagnosed as infantile dislocation of hip join in 54 patients(73 hip join)and treated by expectant treatment as infantile dislocation of hip join.The curative effect of 6 patients were not good,after serious medical examination,careful reading image data,paying attention to curative effect,6 patients were confirmed to be diagnosed as infantile dislocation of hip join and not infantile dislocation of hip join.[Result]The features for diagaosis of congenital infantile coxa vara were:the big greater trochanter of femur and its position moving up,short spacious neck of femur,spacious medial blank in hip joint,superposed neck of femur and lesser trochanter of femur,praeter propter exponent of acetabulum,acceptable succession of Shenton’s line,enlarged angle of femoral head epiphysis,late center of ossification,incorporated abnormity such as breves femoral bone,bowing of femoral bone,abortive clavicle and bones of cranium,equines.[Conclusion]The second center of ossification of femoral head is appeared 6～9 monthes after birth,and it is difficult to discriminate congenital infantile coxa vara and infantile dislocation of hip join before this.Early to realize symptom,physical sign and character of image of congenital infantile coxa vara is significant for correct diagnosis and can avoid misdiagnosis and mistreatment.

Objective To study the medicine dynamics, distribution in tissue and abdominal cavity fluid concentration of 5-FU after giving intraperitoneal by using a gelatin carrier to be made 5-FU slowing-release microballoons. Methods 5-FU slowing-release microballoons medicine release speed, tissue distributing and the concentration in abdominal cavity fluid were measured by high performance liquid chromatography. Results 5-FU wrapped by gelatin were slowly released. The concentration in abdominal cavity fluid was obviously higher than that in tissue or in blood. Using established standard curve line, it was proved that in body area under curve (AUC) of 5-FU slowing-release microballoons group was obviously higher than that of simple 5-FU injection group, analyzed by 3p97 pharmacokinetic software management. Conclusion 5-FU enwrapped by gelatin can retain an effective anticancer activity concentration in abdominal cavity 7 days after giving intraperitoneal and it is distributed mostly in abdominal cavity.

Objective:To study the effect of childhood abuse on student's suicide ideation and attitude to suicide. Methods:90 Technical Secondary School Students with history of being abused and 90 without that history were investigated by applying Questionnaire of Commit Suicide Attitude and Beck Scale for Suicide Ideation. Results:Compared with those without being abused, the abused students had higher percentages having suicide ideation in the past week (14.4% vs. 2.2%,?~2=7.27, P

Objective: To study the effect of childhood abuse on students mental health and alexithymia, and to analyze the relationship between psychosomatic symptoms and alexithymia. Methods: 90 abused Technical Secondary School Students and 90 no-abused Technical Secondary School Students from Xuzhou were investigated by applying Symptom Checklist 90 ( SCL - 90 ) , Toronto alexithymia Scale ( TAS) . Results: Compared with no-abused students, the students who had been abused in childhood showed significantly higher psychiatric symptoms of somatization (1. 8?0. 6) , interpersonal relationship (2.0?0.7) , depression (1.9?0.6) , anxiety (1. 8?0. 5 ) , hostility (1. 8?0. 6) , paranoia (1. 9?0. 6) , psychoticism (1. 8?0. 6) , obsessive (2. 0?0. 6) , phobia (1. 7?0. 6) than control (1.4?0.5, 1.7?0.6, 1.6?0.6, 1.6?0.5, 1.5?0.5, 1.6?0.5, 1.6?0.5, 1.8?0.6, 1.5?0.5, P

Objective To investigate the value of emergency hysterectomy on arresting severe postpartum hemorrhage Methods A retrospective analysis was performed on 10 cases of hysterectomy by reason of postpartum hemorrhage between Jan 1990 and Dec 2000 Results All the patients were severe postpartum hemorrhage that couldn't be controlled by conservative treatment. Among them,there were 5 cases of DIC,2 cases of placental adherence,1 case each of uterine atony,uterine rupture and infection of uterine incision. Blood loss averaged 2 427 ml. All the 10 patients were recovered. Emergency hysterectomy was related to times of gestation and delivery and also to delivery pattern Conclusion Emergency hysterectomy is one of the effective methods on arresting severe postpartum hemorrhage.

Background and purpose: Primary intracranial epithelioid haemangioendothelioma (EH) was rare. This study was to discuss the clinical pathological features, radiographic findings, treatment and prognosis of primary intracranial EH combined with literature review. Methods: We reviewed a case of EH reported from our hospital, and comprehensively analyzed the related literature. Results: Based on our report and review of the literature, EH is rare and with borderline or uncertain behavior. The original tumors demonstrated specific image features. The tumor usually appeared as a small nest or cords composed of eosinophilic epithlioid or spindled cells.Immunohistochemical assay were positive for endothelial markers CD31, CD34 and FⅧ. EH may be treated with complete surgical resection whenever possible and is sensitive to radiation. The EH has a favorable prognosis. Conclusion: EH has variable malignant potential, and should be differentiated from meningioma. Total resection and close follow-up is recommended. Additional radiotherapy is advised for residual tumors.

Objective To study the anti-inflammatory effects of idazoxan (IDA) on endotoxin lipopolysaccharide (LPS) challenged mice in vivo and activated macrophages in vitro,and explore its potential molecular mechanisms.Methods To do the experiments in vivo,30 adult male C57BL/6 mice were randomly divided into control group,model group,and low,medium and high doses IDA groups (IDA-L,IDA-M,and IDA-H groups),n =6 in each group.The inflammatory model was reproduced by intraperitoneal injection of LPS 10 mg/kg,and the control group was injected with the same amount of normal saline.The IDA groups received LPS (10 mg/kg) and IDA 0.3,1.0 and 3.0 mg/kg,respectively.The blood samples of mice in each group were collected at 6 hours after the reproduction of the model.For the in vitro experiments,primary peritoneal macrophages were collected from 20 adult male C57BL/6 mouse cells and they were divided into control group,LPS group (10 mg/L) and LPS+IDA-L,IDA-M,IDA-H groups (10 mg/L LPS + 5,25,100 μmol/L IDA,respectively).Cell culture supernatants were collected at 24 hours after the reproduction of the model.Detection methods:enzyme linked immunosorbent assay (ELISA) was used to determine the levels of serum tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),monocyte chemotactic protein-1 (MCP-1) and nitric oxide (NO).Western Blot was used to determine the effect of IDA on the expression levels of nuclear factor-κB (NF-κB) in macrophages.Results ① For the in vivo experiment,the serum levels of TNF-α and IL-6 were significantly elevated in the model group as compared with those in the control group [TNF-o (ng/L):403.96 ± 40.98 vs.17.50 ± 8.68;IL-6 (ng/L):61 400.31 ± 7 826.61 vs.2 436.30 ± 448.89;both P ＜ 0.01].IDA treatment could inhibit the elevation of inflammatory cytokines in a dose-dependent manner,with the most significant decrease in LPS+IDA-H group [TNF-α (ng/L):170.09 ± 28.53 vs.403.96 ± 40.98,IL-6 (ng/L):16 570.81 ± 1 083.65 vs.61 400.31± 7 826.61;both P ＜ 0.01].② For the in vitro experiment,the levels of TNF-α,IL-6,MCP-1,and NO secreted by LPS-stimulated macrophages were distinctly higher in the LPS group than those in the control group [TNF-α (ng/L):7 259.14 ± 320.70 vs.28.50±27.08,IL-6 (ng/L):14809.60±5852.73 vs.1 113.47±465.53,MCP-1 (ng/L):20847.37± 1 788.33 vs.447.37± 395.69,NO (μmol/L):1 900.00 ± 144.31 vs.603.03 ± 102.18;all P ＜ 0.01].However,IDA intervention could lower the secretion of TNF-α,IL-6,MCP-1 and NO in a dose-dependent manner,with the most notable decrease in the LPS+IDA-H group [TNF-α (ng/L):784.40±281.90 vs.7259.14±320.70,IL-6 (ng/L):1 802.96± 1 534.18 vs.14 809.60± 5 852.73,MCP-1 (ng/L):2005.26± 1 534.28 vs.20847.37 ± 1 788.33,NO (μ mol/L):654.54± 150.21 vs.1 900.00 ± 144.31;all P ＜ 0.05].In addition,IDA at the concentration of 100 μmol/L could promote the translocation of NF-κBp65 in macrophages into the nucleus 15 minutes early and lead to increased NF-κBp65 expression (gray value:18.70 ± 2.29 vs.1.09 ± 0.36,P ＜ 0.05),hut significantly reduce the expression levels of NF-κBp50 in the nucleus at 45 minutes after treatment (gray value:1.99 ± 0.14 vs.2.94 ± 0.54,P ＜ 0.05).Conclusions IDA could significantly reduce inflammation of mice challenged with LPS and inhibit inflammatory cytokines and mediators secreted by macrophage in a dose-dependent manner.High concentration of IDA (100 μmol/L) exhibited the greatest anti-inflammatory effects.The anti-inflammatory effect of IDA may be worked through NF-κB signaling pathway.

AIM: To study the expression of hepatitis B virus core gene in Pichia pastoris and to obtain high-level expressed recombinant HBcAg with good immunoreactivity and high specificity. METHODS: HBV core gene was amplified by PCR from plasmid pHBV1 which contained HBV whole DNA sequence. The PCR product was cloned into pGEM-T vector by TA cloning strategy. After confirmed by DNA sequence analysis, the gene of interest was inserted into the yeast expression vector pPIC9. The recombinant plasmid pPIC9-cAg was constructed and transformed into GS115 by electroporation. The recombinant yeast GS115 was induced by 0.5% methanol. The expressed product was analysed by SDS-PAGE,Western blot and ELISA. RESULTS: The restriction analysis and DNA sequence analysis proved that HBV core gene had already been cloned to yeast expression plasmid pPIC9. The expressed HBcAg existed in SDS-PAGE. Good immunoreactivity and high specificity of the recombinant HBcAg have been proved by ELISA and Western blot. The titre of the recombinant HBcAg in the cell lysate was 1∶ 12 800 . CONCLUSION: The recombinant plasmid pPIC9-cAg was successfully constructed. The recombinant HBcAg with good immunoreactivity and high specificity was successfully expressed in Pichia pastoris expression system and can be applied to further developing HBcAb immunoassay. [