Objective To compare the effectiveness of two different modes of follicular aspiration(FA)guided by transvaginal ultrasound(TVU)in infertile patients with clomiphene citrate(CC)-resistant polycystic ovary syndrome(PCOS).Methods From December 2005 to December 2007,85 patients with CC-resistant PCOS were randomly divided into two groups.Group Ⅰ(thorough treatment)had the every possible small follicles being punctured and aspirated,while group Ⅱ(routine operation)only had the larger follicles being operated.These patients were monitored with TVU and testing the steroid hormone levels.The effect of the two methods on the hormone levels and rates of ovulation and pregnancy were analyzed.Results After the treatments,the cumulative rates of ovulation and pregnancy in the group Ⅰ were higher than those in the group Ⅱ 83.3%(25/30)vs 42.1%(16/38),?2=11.903,P=0.001;43.3%(13/30)vs 13.2%(5/38),?2=7.843,P=0.005].Moreover,the levels of luteinizing hormone(LH)and testosterone(T)were decreased significantly after the treatment in group Ⅰ (17.68?3.11)vs(11.58?1.84),t=9.220,P=0.000;(3.41?2.14)vs(1.47?1.29),t=3.937,P=0.000],whereas those in group Ⅱ were not improved markedly (17.76?4.51)vs(16.11?3.53),t=0.229,P=0.062;(3.88?1.54)vs(3.72?1.27),t=0.995,P=0.326].Conclusions Thorough treatment that has the every possible small follicle being punctured and aspirated can significantly improve the endocrinological abnormalities,and increase the rates of ovulation and pregnancy in patients with CC-resistant PCOS.

Objective:To study the effect of the association of ultrasound and microbubble with short hairpin RNA (shRNA) on the expression of survivin mRNA and protein in cervical cancer (HeLa) cell lines.Methods:Three survivin-shRNA expression vectors (pSIREN/S1/S2/S3) were constructed and introduced into HeLa cells.A corresponding negative control vector was constructed (pSIREN/con).The shRNA vectors were added to Hela cells transfected by LipofectamineTM (P+L) or followed by ultrasound exposure with SonoVue microbubble (US+SO).Blank control group,pSIREN/S1 plasmid only,ultrasound exposure only,pSIREN/S1 and ultrasound exposure were used as controls,respectively.The expressions of survivin mRNA and protein were detected by RT-PCR and western blot analysis,respectively.Results:Three recombinant plasmids were successfully constructed.The plasmids were showed to inhibit the expression of survivin gene mRNA and protein efficiently and specifically except for the pSIREN/con,while pSIREN/S3 was significantly lower than other groups (P

Objective To investigate the gene silencing,apoptosis induction and the suppression of proliferation in vivo transfected by UTMD techniques associated with shRNA techniques. Methods The survivin-shRNA expression vector was constructed. Nude mice were randomly arranged into 3 groups:control group, plasmid injection and ultrasound (P + US), P + UTMD group. Histological examination were evaluated. Protein expressions of Survivin and proliferating cell nuclear antigen (PCNA), Bcl-2, Bax,Caspase-3, Ki-67, nucleostemin (NS), p53 were investigated by immunohistochemistry. Results In transplanted tumors experiment, comparing with those in C and P + US groups, protein expressions of PCNA,Ki-67,Bcl-2, Survivin, NS were down-regulated markedly, while those of Bax, Caspase-3 and P53 were up-regulated significantly ( P ＜ 0.05). Conclusions UTMD combined with shRNA technique can induce apoptosis and inhibit proliferation significantly, without causing any apparently adverse effect,representing a new,promising technology that can be used in the tumor gene therapy and research.

Objective To estimate the diagnostic value of the patients of intrauterine pregnancy abortion (IUPA) of pregnancies of unknown location (PUL) with transvaginal power Doppler ultrasound (TV-PDU) combining Logistic regression model and receiver operating characteristics (ROC) curve.Methods Endometria of PUL were detected by TV-PDU during the early pregnancy.Serum β-HCG and progesterone were analyzed meanwhile.All variables were researched by multiple factors Logistic regression.Logistic regression equationg were established.ROC curve was constructed and the cut-off value of endometrial mnographic findings and clinical data to diagnose intrauterine pregnancy abortion were studied.The accuracy of every univariate and multivariable were evaluated and the validity and stability of the Logistic regression equation were tested.Results The RI value and PSV were included in the multivariate model 1 (M1),while the serum progesterone,type of endometrial resonance and gestation age were included in the multivariate model 2(M2).The area under the curve of M1,M2 were 0.887,0.976,respectivdy.Youden index and positive predictive value were 90.8%,87.5% and 12.213,8.000,respectively.Specificity and positive predictive value of M2 was lower than M1 which had a hisher accuracy.Diagnostic value were increased obviously by the use of M1 and M2 comparing with univariate and the stability was all righL Conclusions Possibility of IUPA could be predicted by TV-PDU simply,atraumaticly and efficiently.A close rehaion is found between the hemodypnmic parameter of endometria and the IUPA.

Objective To elucidate the relationship between quality of life (QOL) and disease activity of systemic lupus erythematosus (SLE),as well as to reveal the factors impacting disease activity and the QOL of SLE utilizing the combination of SLE-specific quality of life questionnaire (SLEQOL) and the medical outcomes survey short form 36 (SF-36).Methods SLEQOL and SF-36 health survey questionnaire were applied.Information on gender,disease duration,age,education level were collected.Serum complement (C3 and C4) level and erythrocyte sedimentation rate (ESR) were measured.Patients were divided into inactive,mild active,moderate active and severe active respectively according to SLE disease activity index (SLEDAI).Pearson's product moment correlation coefficient was used to analyze the correlation between the activity of disease and the QOL.Multiple linear regression was employed to explore the factors which could impact on SLEQOL.Results The physical function of SLEQOL was positively correlated with SLEDAI (r=0.36; P＜0.05).The association between reported health transition of SF-36 and SLEDAI was positive (r=0.19; P＜0.05).Physical functioning,role-physical,role-emotional,body pain and vitality were all negatively correlated with SLEDAI (r:-0.20,-0.19,-0.19,-0.19,-0.21 respectively; P＜0.05).The scores of patients with severe disease activity were significantly increased in the physical functioning of SLEQOL than other three groups (18±10 vs 11±5,12±6,13±7; P＜0.05).Scores on the Health transition of patients with moderate and severe disease activity were lower than those of whom with mild disease activity (23±28.14±17 vs 34±39,P＜0.05).Patients with mild or severe disease activity had lower score than those of patieuts in disease inactive (30±41,34±39 vs 44±44,P＜0.05).Multiple linear regression analyses showed that disease duration and education level might be the influencing factors of SLEQOL.Conclusion QOL of patients with SLE is related to the level of disease activity and is impacted by disease duration and education.

Objective:To transfect genes using ultrasound targeted microbubble destruction (UTMD) techniques and observe the effects of RNA interference on cervical cancer (HeLa) cell line in silencing survivin gene and inducing apoptosis. Methods: Recombinant expression plasmid of short hairpin RNA (shRNA) targeting survivin gene was constructed. It was co-treated with microbubbles and transfected to cultured HeLa cells followed by exposure to ultrasound (P+UTMD group). Moreover, blank control group (C), plasmid group (P), ultrasound exposure group (US), plasmid and ultrasound exposure group (P+US), plasmid+ Lipofectamine group (P+L) were used as controls, respectively. Transfection efficacy was evaluated by observing the red fluorescence in the cells by fluorescent microscopy and flow cytometry(FCM). Ultrasound intensity and exposure time were optimized. Cell apoptosis was investigated using flow cytometry analysis, Hoechst staining, and DNA ladder method. Expression of survivin mRNA was assessed by RT-PCR. Results: Restrictive enzyme digestion and sequencing analysis verified that the recombinant plasmid was successfully constructed. UTMD significantly increased gene transfection efficacy in cultured HeLa cells (P<0.01). Gene transfer was the most prominent at ultrasound intensity of 1.0 W/cm2 and exposure time of 3 min (P<0.01). RT-PCR showed that the expression of survivin mRNA in P+UTMD group was inhibited by (83.33±2.73)%. The differences were significant compared with any other groups (P<0.01). FCM analysis showed that the apoptosis ratio in P+UTMD group was significantly increased as compared with other groups (P<0.01). Hoechst staining and DNA ladder showed that apparent apoptosis and DNA ladder were detected only in P+UTMD and P+L groups. Conclusions:UTMD effectively enhances the transfection efficacy of expression plasmid. It is a novel and effective non-viral gene transfer system and has promising foreground. UTMD mediates RNA interference silenced survivin gene and induces significant cell apoptosis, which provides a new method for tumor research and gene therapy.

Objective To explore effects of hcmoperfusion on toxic ingredients in plasma of rabbiis with a-cute intoxication of Radix Aconiti Kusmezoffii Monkshood.Method Sixteen male Japanese Giant Ear Rabbits were randomly divided into acute poisoning(AP)group and acute poisoning + hemoperfusion(AH)group(8 an-hnals in each group).Acute poisoning models were established in rabbits of both groups with intragastric adminis-tration of Radix Aconiti Kusmezoffii Monkshood liquid in dose of 1 mL/kg in order to produce arrhythm which oc-curred within ode hour after intragastric administration was regarded as the criteria of successful animal model.and then hemoperfusion with active carbon was performed for 2 hours in AH group.The pathological chanses of brain,myocardium and hepatic tissues were observed.The plasma concentrations of toxicants including mesaconitine,a-conitine and hypaconitine were measured by using HPLC-MS at 1 h,2 h,3 h,and 6 h after poisoning.Student's T test was used to identify the significance.Results The brain.myocardium and hepatic tissues of the rabbits in AP group showed hyperemia and edema which were attenuated after hemoperfusion.The plasma concentrations of mesaconitine,aconitine and hypaconitine revealed no significant differences between AP group and AH group with-in one hour after poisoning(P＞0.05),while at 2 h and 3 h after poisoning,the plasma concentrations of mesaconitine were(2.11±1.08)ng/mL,(2.02±1.46)ng/mL,respectively,aconitine(39.70±9.31)ng/mL,(19.71±16.06)ng/mL,respectively,and hypaconitine(1.70±0.71)ng/mL,(2.12±1.33)ng/mL,respec-fively in AH group,and they were significantly lower than those in AP group(P＜0.05).Conclusions The the plasma concentrations of mesaconitine,aconitine and hypaconitine were lower and the histopathological changes were attenuated after hemoperfusion.Hemoperfusion is a good intervention for acute intoxication of Radix Aconiti Kusmezoffii Monkshood.

Aim To observe the antiviral function of Radix Notoginseng flavone on a virus infection model caused by COXB_3 virus in vitro and in vivo. Methods To establish an experimental in vitro model of viral myocarditics based on primary culture of myocardial cells of newborn Wistar rats and a Balb/c mouse model of viral myocarditics through intraperitoneal injection. Results In vitro Radix Notoginseng flavone obviously inhibited pathologic change of cultured cardiac cells; it reduced release of enzymes from myocardial cells, and increased the level of interferon in mouse spleen, and inhibited inflammatory cells infiltration among cardiac cells in Balb/c mice infected by COXB_3 virus; survival rate of mice in treated groups increased. Conclusion Radix Notoginseng flavonoe has therapeutic efficacy on viral myocarditis caused by COXB_3 virus.

To study the role of the thyroid hormone receptor TRalphaA involved in the process of the metamorphic development of Japanese flounder, we firstly cloned the TRalphaA gene, then ligated into the fusion expression vector pET30a and expressed in Escherichia coli DE3 (BL21) host cells. After induced for 4 h with 1 mmol/L Isopropyl beta-D-Thiogalactoside, the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. The recombinant protein was denatured and purified by His-Bind resin, then renatured through gradient washing on His-bind resin column. After that, polyclonal antibody was prepared by immunizing New Zealand rabbits with purified protein. Dot blotting analysis showed the antibody with the titer of 1:200 000 reacted specifically to the expressed recombinant protein. Furthermore, a chromatin immunoprecipitation assay was performed to identify the specific binding between the antibody and TRalphaA in living cells of Japanese flounder. The result showed that thyroid hormone was involved in the alkaline phosphatase (ALP) gene transcriptional regulation through TRalphaA in vivo.
Alkaline Phosphatase ; genetics ; immunology ; Animals ; Antibodies ; immunology ; Escherichia coli ; genetics ; metabolism ; Flounder ; physiology ; Metamorphosis, Biological ; physiology ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Thyroid Hormone Receptors alpha ; biosynthesis ; genetics ; immunology

Purpose To analyze the clinicopathologic and immunohistochemical features, differential diagnosis and prognosis of papil-lary renal cell carcinoma (PRCC). Methods Thirty-two cases of PRCC diagnosed were reviewed. A retrospective study was per-formed including reviewing the clinical documents, pathological sections and immunohistochemical stainning and follow-up was made of 32 cases of PRCC. Twenty-one patients were treated with radical nephrectomy, eleven patients were treated with partial nephrectomy. Results Among 770 cases of renal epithelial tumors 32(4. 2%) cases of PRCC were detected. Histologically, the PRCC were charac-terized by varying proportions of papillary and tubular architecture covered by single or multiple layer of tumor cells with scanty or volu-minous basophilic or eosinophilic cytoplasm. Foam cells and psammoma bodies were seen in some papillary cores and stroma, and the cytoplasm of some tumor cells contained hemosiderin. Of these 32 patients, 18 and 14 were diagnosed type-Ⅰand type-IIPRCC, re-spectively. Type-I, with small cuboid cell and pale cytoplasm, 16 of them were low in Fuhrman grading, Type-II, with large colunmar cells, rich in eosinophilic cytoplasm, 12 of them were high in Fuhrman grading. Immunohistochemically, the PRCC showed positive immunostaining for vimentin, EMA, CK(AE1/AE3), CK7, CD10 and AMACR. All the tumors studied were negative for CK (34βE12) and TFE-3. Follow-up data were available for 31 cases, 4 patients died of cancer specific causes, 1 with type-Ⅰand 3 with type-II tumors after surgery. The other 27 patients were alive without recurrence or metastasis. High Fuhrman grading, intravascular tumor emboli, lymph node metastasis and high clinical stage were prognostic indicators in PRCC. Conclusions PRCC with unique pathological features is not a common subtype of renal cell carcinoma in China. The presence of higher nuclear grade, sarcomatoid ele-ments or clear cell carcinoma structure may indicate an aggressive biologic behavior and poor prognosis. Close attention to the cytologic and growth pattern characteristics will allow us to arrive at the proper diagnosis in most cases, although sometimes immunohistochemis-try and rarely molecular genetic evaluation may be needed.