Aged ; Carcinoma, Squamous Cell ; pathology ; surgery ; Cholecystectomy ; Gallbladder Neoplasms ; pathology ; surgery ; Humans ; Male

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.

ObjectiveTo study the imaging characteristics of SPN of adenocarcinoma (ASPNs) on 18F-FDG PET/CT.MethodsThe morphological and metabolic features of 35 ASPNs on FDG PET/CT were retrospectively reviewed.SUVmax (SUV) was measured and ΔSUVmax was calculated according to ΔSUVmax =(SUVmax on delay imaging - SUVmax on early imaging)/SUVmax on early imaging × 100％.Statistical analysis was performed by software SPSS 11.5 using t-test,analysis of variance and Fisher exact test.Results( 1 ) Fifteen ASPNs (42.86％,15/35) presented as nodular pattern on FDG PET imaging,while 20 (57.14％,20/35) as lamellar,cloudy or ill-defined patterns.The SUVmax of these ASPNs followed a descending order of nodular,lamellar,cloudy and ill:defined on both early and delay imaging (F =30.696 and 24.758,both P＜0.001).(2)There were 54.29％ (19/35) ASPNs with SUVmax ≥2.5 and 45.71％ (16/35) ASPNs with SUVmax ＜2.5.(3) Of 35 ASPNs,24(68.57％ ) were solid nodules and 11(31.43％) were ground glass nodules with SUVmax =4.54 ±2.69 and 1.30±0.87,respectively (t =-5.234,P ＜O.001 ).(4) The SUVmax of ASPNs on delay FDG imaging (4.22 ±3.52) was significantly higher than that on early imaging (3.49 ±2.72) (t =-4.021,P ＜0.001 ).However,SUVmax was dependent on SUVmax on the early imaging:when SUVmax ≥2.5,ΔSUVmax was positive in 94.74％ (18/19) of ASPNs; while SUVmax ＜2.5,ΔSUVmax was positive in 56.25％ (9/16) of ASPNs (P =0.013).(5) Of 31 ASPNs with cell differentiation data,there were 10/17 well-differentiated ASPNs and 13/14 poorly-differentiated ASPNs with positive ΔSUVmax ( P =0.045 ).The average SUVmax of well-differentiated ASPNs was significantly lower than that of poorly-differentiated ASPNs ( 1.70 ± 1.51 vs 4.91 ± 2.69,t =- 3.951,P ＜ 0.001 ).Conclusions The morphological and metabolic features of ASPNs are diversified.It is common for ASPN to present with SUVmax ＜ 2.5.ΔSUVmax may be helpful for differentiating malignant from benign SPNs.

Objective To construct recombinant adenovirus Ad/MDC-VP1 and investigate its im-muno-boosting effect of the mice primed with the experimental DNA vaccine against Coxsackievirus infection. Methods The recombinant adenovirus Ad/MDC-VP1 was constructed and packaged. The Western blot analysis was used to verify the target protein. BALB/c mice were divided into four groups: Ad/MDC-VP1 group, pcDNA3/MDC-VP1 group, pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost group and PBS group. The mice in each group were immunized intramuscularly. The titers of serum IgG and neutralizing antibody were tested by ELISA and trace neutralization assay, respectively. The lymphocytes proliferation activity and specific CTL cytotoxic activity were tested by CCK-8 assay. The mice in each group were challenged with le-thal dose of Coxsackievirus, and the assay of the serum virus titers and the observation of protection efficacy against Coxsackievirus infection were carried out. Results The recombinant adenovirus Ad/MDC-VP1 was successfully constructed and the target protein was expressed. It was observed that the titers of CVB3 VP1 specific antibody, lymphocyte stimulation index, CTL cytotoxicity activities and protection rate of the pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost group were much higher than those of the rest groups( P < 0.05), and the titer of serum virus was lower after CVB3 challenged ( P < 0.05 ). Conclusion Both the cellular and humoral immune responses in mice could been significantly enhanced by the pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost strategy.

Objective To compare the immune effects of Coxsackievirus B3 (CVB3) capsid protein VP1 expressed bacterially, recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1which express VP1 protein in mice. Methods After expressed in prokaryotic cells, VP1 protein was purified. Recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1 were amplified and extracted. Six to 8-week-old, male BALB/c mice were divided into four groups randomly. Each group contained 18 mice. The mice of pcDNA3/VP1 group or VP1 protein group were immunized intramuscularly with three injections at three weeks apart, of recombinant plasmid pcDNA3/VP1 at a dose of 100 μg/mouse or recombinant protein VP1 at a dose of 50 μg/mouse. The mice of rAd/VP1 group were immunized intramuscularly twice at two weeks interval with rAd/VP1 at a dose of 1.2 × 107 PFU. The control group was mock-immunized with 100 μl of PBS intramuscularly. Mice were bled from the retroorbital sinus plexus every two weeks after each immunization. ELISA and micro-neutralization test were used to detect levels of CVB3-specific IgG antibody and neutralizing antibody titers in the sera of immunized mice. Three weeks after the last immunization, the cytotoxic T lymphocyte(CTL) killing activity of spleen lymphocytes was detected with CCK-8 assay. Subsequently, virus titers in the sera of immunized mice were determined by the 50% cell culture infective dose( CCID50 ) assay on HeLa cell monolayers and percentage of animals surviving were observed after lethal CVB3 attack over a period of 21 days. Results The titers of specific IgG antibody and neutralizing antibody in sera of VP1 protein immunized mice were higher than other groups( P ＜0.05 ). While CTL killing activity of spleen lymphocytes of VP1 protein immunized mice was lower than mice in rAd/VP1 group( P ＜0. 05). Virus titers in sera of VP1 protein immunized mice were lower than the mice in pcDNA3/VP1 or rAd/VP1 groups ( P ＜ 0.05 ), while survival rate was significantly higher than these two groups ( P ＜ 0.05 ).Conclusion VP1 protein induced higher level of humoral immune response and acquired obvious immune protection effects in mice. The immunizing potency of VP1 protein vaccine surpassed plasmid pcDNA3/VP1or recombinant adenovirus rAd/VP1. It appeared to be a promising candidate among the three different vaccines.

Objective To study the genotype distribution of insulin receptor substrate-2(IRS-2)gene 1057G/A polymorphism in Han population from Southwest China,and to explore its association with the metabolism of glucose and lipids,insulin resistance and islet?-cell function in type 2 diabetic patients and subjects with impaired glucose tolerance(IGT).Methods A total of 929 Hans[462 patients with type 2 diabetes(DM group) and 164 subjects with IGT(IGT group)and 303 normal controls(NC group)]from Chongqing and nearby regions were screened for 1057G/A polymorphism of IRS-2 gene by PCR-RFLP assay.Body mass index(BMI),plasma glucose,serum insulin and lipid profile,high-sensitive C-reactive protein(hsCRP)and non-esterified fatty acid were measured.Homeostasis model assessment of insulin resistance(HOMA-IR)and disposition index(DI)were used to estimate insulin resistance and?-cell function respectively.Results In DM group,A allele frequency was significantly lower than that in NC group(0.326 vs 0.388,X~2=6.19,P=0.01).Compared with NC group,AA genotype frequeney was lower and GG genotype frequeney was higher in DM group(0.104 vs 0.135 and 0.452 vs 0.360 respectively,X~2=6.80,P

AIM:To establish an effective method for purification and culture of human cumulus cells(CCs) in vitro,and to compare the characteristics between CCs and mural granulosa cells(MGCs).METHODS:Follicular fluid and cumulus complex from the patients undergoing intracytoplasmic sperm injection were collected.CCs were mechanically cut from cumulus complex and then directly inoculated on a Petri dish, and MGCs were obtained from follicular fluid through density gradient centrifugation.The expression of follicle stimulating hormone receptor(FSHR)was determined by immunofluorescence.The cell growth curves were measured by CCK-8 assay.The secretion of estrogen was detected by ELISA.RESULTS:After incubated for 24 h, the adherence of CCs was observed.CCs and MGCs had similar growth characteristics and FSHR expression.The similar cell growth curves were observed by CCK-8 assay and the results of ELISA showed that they had comparable secretion of estrogen.CONCLUSION:Direct culture of CCs mechanically cutting from cumulus complex is an effective method.CCs had similar growth characteristics,growth curves and secretion of estro-gen to MGCs cultured in vitro and could be a substitutive source of granulosa cell subsets.

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.
Cations ; Gene Expression Regulation, Viral ; drug effects ; Hepatitis B virus ; genetics ; Liposomes ; chemistry ; Plasmids ; RNA, Small Interfering ; chemistry ; Trans-Activators ; genetics ; metabolism

[Objective]To evaluate the practicality of non-mydriatic digital fundus camera in the remote screen of diabetic retinopathy for community residents.[Methods]Ninety-two patients(184 eyes)with type 2 diabetes mellitus have been taken 1-field and 5-field non-mydriatic fundus photography and examination in mydriatic fundus by pre-placed-mirror ophthalmoscopy by a ophthalmologist,the results were sent to the hospital with a computer programs. A specialist evaluates the consistency of detectable rate of diabetic retinophathy(DR)among the 3 methods.The time of tak-ing 1-field and 5-field non-mydriatic fundus photography is compared.[Results]All the three methods show good consis-tency in detectable rate of DR compare with each other,the κ value is 0.89 for 1-field non-mydriatic fundus photography and examination in mydriatic fundus by preplaced-mirror ophthalmoscopy,0.95 for 1-field and 5-field non-mydriatic fundus photography and 0.95 for 5-field non-mydriatic fundus photography and examination in mydriatic fundus by pre-placed-mirror ophthalmoscopy,respectively. The 1-field non-mydriatic fundus photography spent less time compares with 5-field non-mydriatic fundus photography(55.4±5.8 vs 405.9±68.5 s,P<0.01).[Conclusion]The remote screen for diabetic retinopathy in community based on non-mydriatic digital fundus camera is worth promoting,we suggest 1-field non-mydriatic fundus photography as a screen method.

Acute Kidney Injury ; etiology ; Fanconi Syndrome ; etiology ; Female ; Humans ; Lymphoma, Non-Hodgkin ; complications ; Middle Aged