Objective: To investigate the expression and expression efficiency of recombinant adeno-associated viral containing human thyrotropin receptor A subunit (rAAV2/9-hTSHR289-IRES-ZS Green) in mouse embryonic fibroblasts (NIH3T3) or in mice. Methods: NIH3T3 was infected by rAAV2/9-hTSHR289-IRES-ZsGreen and its TSHR289 protein expression was detected by Western blotting. Female BALB/c mice were intramuscularly injected with different doses of rAAV2/9-hTSHR289-IRES-ZsGreen; the expressions of the target protein in different organs were determined by immunofluorescence while the serum TSHR antibody (TRAb) titer was determined by radioimmunoassay. Results: NIH3T3 cells transfected with rAAV2/9-hTSHR289-IRES-ZsGreen expressed hTSHR289 protein both at 48 h and 72 h. The expression of target protein at 48 h or 72 h was significantly higher than that at 24 h (P<0.05). There were different levels of TSHR289 expression in leg skeletal muscle, heart, liver and spleen under fluorescence microscope. The results of radioimmunoassay showed that the higher dose injection produced a higher titer of TSHR antibody, but only the TRAb level in high dose and control injection exhibited a statistical difference (P<0.05) at week 4, week 8 and week 12. Furthermore, strong antibody response was observed in the mice injected with high dose and medium dose of rAAV2/9-hTSHR289-IRES-ZsGreen at week 4 and gradually weakened between 4 and 8 weeks. In addition, the antibody lasted for 12 weeks. Results: rAAV2/9-hTSHR289-IRES-ZsGreen can highly express hTSHR289 protein in vivo and in vitro, and the hTSHR289 protein displays strong immunogenicity. It indicates that rAAV2/9-hTSHR289-IRES-ZsGreen might become a more effective means of preparing Graves' disease model.

ABSTRACT:Objective To explore the effects of miRNA-1246 (miR-1246)on cell proliferation,invasion and migration in human cervical squamous cell carcinoma (CSCC)cell line SiHa.Methods SiHa cells were assigned into 3 groups:miR-1246 analog group,miR-1246 antagonist group and control group.Transfection efficiency was determined.The MTT assay,transwell assay and wound healing assay were performed respectively to evaluate the proliferation,invasion and migration abilities of SiHa cells.Western blot was carried out to detect the expression of thrombospondin-2 (THBS2)before and after transfection.A THBS2 3’-UTR-containing dual luciferase plasmid was synthesized and co-transfected with miR-1246 into SiHa cells to observe the luciferase enzyme activity.Results MTT assay,transwell assay and wound healing assay revealed that the abilities of proliferation,migration and invasion were significantly enhanced (P<0.01)in SiHa cells transfected with miR-1246 analog,but suppressed in SiHa cells transfected with miR-1246 antagonist.Western blot showed that SiHa cells transfected with miR-1246 analog had significantly decreased THBS2 expression (gray value = 6 .2 8 ± 1 0 .2 2 , P=0 .0 1 3 ) while those transfected with miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90±19.81, P=0.037).After co-transfected with miR-1246 and THBS2 3’-UTR-containing plasmid,SiHa cells exhibited a decreased level of luciferase enzyme expression.Conclusion miR-1246 promoted the proliferation,invasion and migration of CSCC SiHa cell, and it might promote CSCC tumorigenesis and progression by suppressing the expression of its target gene THBS2 .

Objective To explore the influence of body position intervention combined with lying position logo on the compliance of patients after vitrectomy needing to keep lying position. Methods A total of 100 patients, who were hospitalized in ophthalmology ward from January to June 2013,received vitrectomy combined with gas or silicone oil filled were involved. All the patients were divided into intervention group (60 cases) and control group (40 cases) according to the order of admission. Patients in control group received routine nursing after retinal vitreous operation, guiding the proper way to keep lying position, putting a sponge pillow, shaping凹, under patients′heads. And some pertinence measures were applied in intervention group on this basis. Results The percentage of the patients who could keep the special position complying with doctor's orders at 72 h and 1 week after the operation were 95.0%(57/60),91.7%(55/60) in intervention group, which were greater than those in control group 80.0%(32/40),75.0%(30/40), and there were significant differences,χ2=4.090,4.003,P<0.05.The mastery of the body position, and the satisfaction of the nursing care were 96.7%(58/60), 98.3%(59/60) in intervention group, which were better than those in control group 77.5%(31/40), 85.0%(34/40),and there were significant differences, χ2=7.154, 4.666,P <0.05. Conclusions Developing a reasonable nursing plan depending on the body position intervention and relevant nursing measures can help the patients comply with doctor's orders effectively, improve comfort,and extend the time to keep the special position,so as to improve postoperative recovery and ensure the success of the operation.

Objective To explore the skill of mien)surgical removal and preservation and functional recovery of facial nerve for large acoustic neuromas.Methods The clinical data of 82 consecutive patients with large acoustic neuromas operated upon through the suboccipital retrosigmoid approach were retrospectively reviewed.The cases with HB grade Ⅲ - Ⅵ of facial nerve were divided randomly into two groups,the one group was given conventional therapy and the other group was given Kabato rehabilitation training.Results In those cases,acoustic neuromas were totally removed in 77 (93.9%),subtotally removed in 5.Facial nerve was kept anatomically intact in 78 cases of the patients (95.1%).Recovery of facial nerve function was faster and more effective in the training group than that in the un-training group.During the follow-up ranging from 6 months to 3 years,among 5 cases of subtotally resection,2 cases relapsed.Three patients in our series complained of chronic headache postoperatively.No patient long time coma or died.Conclusion The goal of large acoustic neuromas treatment should be total removal in one stage and preservation of facial nerve function,as they determine a patient's quality of life.Kabat rehabilitation training is the effective method of functional recovery of facial nerve after operation.

Objective To explore the effect of Yes-associated protein 1 (YAP1) and potential mechanism on erlotinib ( ER) resistance in lung adenocarcinoma.Methods In PC-9 cells and acquired ER resistant PC-9 ( PC-9/ER) cells, the expression changes in YAP1 gene were measured by quantitative real-time PCR( RT-qPCR) and Western blot.Indirect immunofluorescence was adopted to observe the location of YAP1.PC-9/ER cells were treated with the verteporfin ( VP, YAP1 inhibitor) for 24 h and 48 h, respectively.Expression changes in mRNA and proteins of YAP1, AKT and p-AKT were detected in the presence or absence of VP.The effect of VP was analyzed by drug resistance index using Cell Counting Kit 8(CCK-8) assay.Results The resistance index of PC-9/ER cells was (99.80 ±25.81).Compared with PC-9 cells, the expression levels of YAP1 mRNA and protein were increased in PC-9/ER.The inhibitory efficiency of VP was (50.96 ±5.86)%, and the levels of AKT and p-AKT proteins were down-regulated by the inhibition of YAP1 simultaneously.The half maximal inhibitory concentration (IC50) of PC-9/ER decreased from (11.10 ±2.72) to (1.47 ± 0.32)μmol/L (P =0.024).Resistance index was reduced to one eighth of the original.Conclusion These results indicate that the YAP1 mediates ER resistance in lung adenocarcinoma.Suppression of YAP1 can reduce the resistance through PI3K/AKT signaling pathway.Therefore, YAP1 may be a potential target for lung cancer gene therapy.

BACKGROUND:Bismth-doped iron nanoparticles modified by hyaluronic acid (HA-BiIOPs) not only act as an effective MRI contrast agent, but also as a radiotherapy sensitizer.OBJECTIVE:To fabricate the HA-BiIOPs and to observe its effect to enhance the radiosensitivity of glioblastoma cells U87MG under X-ray radiation.METHODS:HA-BiIOPs were synthesized using hydrothermal polyol method. (1) Cytotoxicity: A cytotoxicity test was carried out on U87MG cells and rat vascular smooth muscle cells (VSMCs). Cell proliferation rate of two kinds of cells cultured with different concentrations of HA-BiIOPs (0, 12.5, 25, 50, 100, 200, 400 mg/L) at 24 hours after culture were determined by cell counting kit-8 assay. (2) Histological analysis: ICR mice were sacrificed after intravenous injection of HA-BiIOPs, and pathological changes of mouse visceral organs were observed under an optical microscope. (3) Cellular uptake: The HA-BiIOPs after entered into the cytoplasm were observed by Prussian blue staining. (4) Radiosensitization test: U87MG cells at Logarithmic growth stage were cultured in culture medium as control group, subjected to X-ray irradiation (0, 3, 6, 9 Gy) as radiotherapy group, cultured in HA-BiIOPs (0, 12.5, 25, 50, 100, 200 and 400 mg/L) as HA-BiIOPs group or subjected to HA-BiIOPs culture plus X-ray irradiation as combined therapy group. Then, the cell proliferation rate and cloning efficiency were measured at 24 hours after treatment.RESULTS AND CONCLUSION:(1) The HA-BiIOPs at different concentrations were non-cytotoxic for VSMC and U87MG cells. (2) After intravenous injection of HA-BiIOPs, there was no obvious toxicity to the mouse susceptible organs. (3) After 6 hours of culture, the HA-BiIOPs could be internalized by U87MG cells. (4) The proliferation rate of U87 cells was negatively correlated with the concentration of HA-BiIOPs (0-200 mg/L) and X-ray dose (0-9 Gy). Especialy, the combination of 6 Gy X-ray irradiation with 200 mg/L HA-BiIOPs dramatically decreased the cell viability that was decreased to (41±7)%. In the combined therapy group with 6 Gy X-ray and 100 mg/L HA-BiIOPs, the cells proliferation rate was significantly lower than that in the control and radiotherapy groups (P < 0.05). These results indicate that HA-BiIOPs have a radiosensitizative effect on glioblastoma cells U87MG.

OBJECTIVEA multi-center randomized phase III clinical trial was designed to evaluate the efficacy, clinical benefit response (CBR) and toxicity profile of germcitabine (GEM) or GEM plus cisplatin (CDDP) for locally advanced (LAPC) or metastatic pancreatic cancer (MPC).

METHODSFrom July 2000 to May 2001, 42 untreated patients with LAPC or MPC were collected and randomized into two groups: Arm A-GEM 20 patients and Arm B-GEM + CDDP 22 patients. Eligibility criteria were: cytologically and pathologically proven pancreatic carcinoma, Karnosky performance status (KPS) 60 - 80, age 18 - 75 yrs, adequate hematological, renal and liver function, measurable disease, and controllable pain. For Arm A patients, weekly dose of GEM 1 000 mg/m(2)/w for 7 times followed by a week rest. Then weekly GEM at the same dose for 3 times every 4 weeks. Arm B patients were given weekly dose of GEM 1 000 mg/m(2)/w for 3 times every 4 weeks combined with CDDP 60 mg/m(2) on D15 for 3 cycles.

RESULTSThirty-four patients were available for objective response (Arm A 16 and Arm B 18) and 36 (Arm A 16 and Arm B 20) for CBR evaluation. In Arm A and Arm B, PR 1 (6.3%) and 2 (11%), MR 4 (25%) and 3 (16.7%), SD 7 (43.8%) and 8 (44.4%), PD 4 (25%) and 5 (27.8%), PR + MR 31.3% and 27.8%, PR + MR + SD 75% and 72.2% were observed. Positive CBR was 14/16 (87.5%) in Arm A and 14/20 (70.0%) in Arm B. The negative results was 2/16 (12.5%) in Arm A and 6/20 (30.0%) in Arm B. The median time of disease progression was not yet available at present. The 3-month survival rate of both Arm A and B was 100%, the 6-month survival rates of Arm A and B were 81.3% and 61.6% and the 12-month survival rates of Arm A and B was 31.3% and 11.1%, with median survivals of 273 and 217 days. The incidence of hematological and non-hematological toxicity of Arm A was lower than that of Arm B without statistical significance. The toxicity ranging from being mild to moderate was manageable.

CONCLUSIONGEM or GEM plus CDDP is able to lead to a moderate objective response rate, also significantly improve the quality of life in patients with locally advanced or metastatic pancreatic cancer patients, prolonging the survival time with tolerable toxicity.

Adult ; Aged ; Antimetabolites, Antineoplastic ; adverse effects ; therapeutic use ; Antineoplastic Agents ; adverse effects ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; CA-19-9 Antigen ; analysis ; Cisplatin ; adverse effects ; therapeutic use ; Deoxycytidine ; adverse effects ; analogs & derivatives ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; drug therapy ; mortality ; Survival Rate ; Treatment Outcome

Objective Although principal components analysis profiles greatly facilitate the visualization and interpretation of the multivariate data,the quantitative concepts in both scores plot and loading plot are rather obscure.This article introduced three profiles that assisted the better understanding of metabolomic data.Methods The discriminatory profile,heat map,and statistic profile were developed to visualize the multivariate data obtained from high-throughput GC-TOF-MS analysis.Results The discriminatory profile and heat map obviously showed the discriminatory metabolites between the two groups,while the statistic profile showed the potential markers of statistic significance.Conclusion The three types of profiles greatly facilitate our understanding of the metabolomic data and the identification of the potential markers.

Objective To investigate the short-term efficacy of dapagliflozin in the treatment of non-diabetic patients with severe aortic stenosis after transcatheter aortic valve replacement(TAVR).Methods A total of 84 non-diabetic patients with severe aortic stenosis after TAVR who were admitted to Zhengzhou Cardiovascular Hospital from March 2019 to September 2022 were selected as research subjects.According to the postoperative treatment,the patients were divided into control group and observation group,with 42 patients in each group.Patients in both groups underwent TAVR.The patients in the control group were given routine treatments such as antiplatelet drugs,cardiac remodeling improvement drugs,and diuretics after TAVR;patients in the observation group were given dapagliflozin 10 mg daily for 6 months in addition to treatment in the control group.The left ventricular ejection fraction(LVEF),left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),aortic valve peak gradient(AVPG)and aortic valve peak velocity(AVPV)of patients in the two groups were measured by using an ultrasound diagnostic instrument before surgery,3 days and 6 months after TAVR;before surgery and 6 months after the TAVR,low-density lipoprotein cholesterol(LDL-C)in serum of patients in the two groups was detected by direct measurement method,lipoprotein a[Lp(a)]level in serum was detected by latex agglutination reaction method,hypersensitive C-reactive protein(hs-CRP)level in serum was detected by rate scattering turbidimetry;the levels of N-terminal pro B-type natriuretic peptide(NT-proBNP),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and interleukin-1 β(IL-1 β)in serum were detected by using enzyme-linked immunosorbent assay;the glycated hemoglobin level of patients in the two groups was measured by ion exchange chromatography.Results There was no statistically significant difference in LVEF,LVESD and LVEDD of patients in the two groups before and 3 days after surgery(P＞0.05);after 3 days of surgery,the AVPG and AVPV of patients in the two groups were significantly lower than those before surgery(P＜0.05).Six months after surgery,the LVEF of patients in the two groups was significantly higher than that before and 3 days after surgery,while LVESD,LVEDD,AVPG and AVPV were significantly lower than those before and 3 days after surgery(P＜0.05).There was no statistically significant difference in LVEF,LVESD,LVEDD,AVPG and AVPV between the control group and the observation group before and 3 days after surgery(P＞0.05).After 6 months of surgery,the LVEF of patients in the observation group was significantly higher than that in the control group,while LVESD and LVEDD were significantly lower than those in the control group(P＜0.05);there was no statistically significant difference in AVPG and AVPV of patients between the observation group and control group(P＞0.05).Before surgery,there was no statistically significant difference in body mass index(BMI),LDL-C and Lp(a)of patients between the two groups(P＞0.05).Six months after surgery,the BMI,LDL-C and Lp(a)of patients in the two groups were significantly lower than those before surgery,and the BMI,LDL-C and Lp(a)of patients in the observation group were significantly lower than those in the control group(P＜0.05).Before surgery,there was no statistically significant difference in the hs-CRP,NT-proBNP,IL-6,TNF-α and IL-1 β of patients between the two groups(P＞0.05);six months after surgery,the hs-CRP,NT-proBNP,IL-6,TNF-α and IL-1 β of patients in the two groups were significantly lower than those before surgery,and the hs-CRP,NT-proBNP,IL-6,TNF-α and IL-lβ of patients in the observation group were significantly lower than those in the control group(P＜0.05).There was no statistically significant difference in glycated hemoglobin of patients between the two groups before and six months after surgery(P＞0.05),and no statistically significant difference in glycated hemoglobin of patients in the two groups six months after surgery compared with that before surgery(P＞0.05).Conclusion Dapagliflozin can effectively improve cardiac structural remodeling,regulate lipid metabolism,reduce the expression of inflammatory factors and promote the recovery of heart function in non-diabetic patients with severe aortic stenosis after TAVR.

Objective:To study the basic situation of Yunnan Province Suncus murinus carrying plague phage and to explore its epidemiological significance. Methods:From 2015 to 2018, a survey of plague host animals was carried out in 10 investigation sites in the historical plague foci, new plague foci (after 1982) and stubborn plague foci of domestric mouse in Yunnan Province. The plague phage was isolated and cultured from the intestinal specimens of Suncus murinus. The growth of plaque was observed by double-layer plate method, and the morphology and structure of plague phage were observed under electron microscope. At the same time, intestinal samples were taken to detect the structural gene caf1 of F1 antigen of Yersinia pestis. Results:In this study, a total of 157 Suncus murinus were captured and 16 strains of plague phage were isolated, with a total isolation rate of 10.19%. There was no difference in plague phage isolation rate between historical plague foci (10.00%, 1/10) and stubborn plague foci (16.22%, 12/74), new plague foci (4.11%, 3/73, χ 2 = 0.00, P = 0.965; Fisher test, P = 1.000). However, there was a difference in plague phage isolation rate between stubborn plague foci and new plague foci (χ 2 = 5.88, P = 0.015). There was no significant difference in the isolation rate of plague phage among different sex, growth period and habitat ( P > 0.05). The plaque morphology of the isolated plague phage was diverse, of which four strains were myotavirus phages; and all samples were negative for F1 antigen structural gene caf1. Conclusions:Suncus murinus is widely distributed in the domestic mouse plague foci in Yunnan Province, and the animals carry a certain number of plague phage. Regular surveillance of Suncus murinus and their plague phage has a certain guiding significance for the surveillance and early warning of plague in Yunnan Province.