Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Squamous Cell ; drug therapy ; radiotherapy ; Combined Modality Therapy ; Esophageal Diseases ; etiology ; Esophageal Neoplasms ; drug therapy ; radiotherapy ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Particle Accelerators ; Platinum Compounds ; administration & dosage ; Radiation Injuries ; etiology ; Radiotherapy, High-Energy ; adverse effects

Objective To construct the pseudovirus containing nucleic acid(NA)fragments of Marburg virus,Zaire Ebola virus,Sudan Ebola virus,Lassa fever virus and Yellow fever virus by using a lentiviral vector system in order to provide a reference standard for the detection of the five viruses.Methods The gene fragments of the above five viruses were synthesized in vitro,connected into a single gene by fusion PCR technique,and cloned into lentiviral vectors with its auxiliary vector.After co-transfecting into 293T cells,the supernatants were collected on 48 h and 72 h post transfection. The naked NA was cleaned from the supernatants using DNase and RNase digestion before pseudotype virus was purified and concentrated.After the NA of the pseudotype virus,were extracted normal PCR and real-time PCR were conducted. Results Sequence analysis showed that the five target genes in vitro synthesis were properly connected and inserted into lentivirus vectors.Using the NA of the pseudotype virus as the template,both normal PCR and real-time PCR could sensitively amplify the target gene with the primers and probes of the above five,viruses respectively.The result indicated that the pseudovirus particles containing the five kinds of hemorrhagic fever virus target genes were successfully packaged. Conclusion The pseudovirus particles containing gene fragments of five viruses are constructed,which can be used as a common reference standard for NA detection.

Adult ; Diagnosis, Differential ; Female ; Glycogen ; metabolism ; Glycogen Storage Disease Type II ; pathology ; Humans ; Liver ; metabolism ; pathology ; Muscle, Skeletal ; metabolism ; pathology ; Muscular Dystrophies, Limb-Girdle ; pathology ; Pyomyositis ; pathology ; Young Adult

Objective To investigate the daily total fluoride intake in relating to the prevalence of skeletal fluorosis in two villages in Jiangsu Province,in order to provide the scientific evidences for the control and prevention of endemic fluorosis.Methods Adults sampled from a high-fluoride Village,Wamiao,and a low-fluoride Village,Xinhuai,were surveyed in this study according to the fluoride concentration in their household shallow well.The average daily total fluoride intake from difierent sources and the skeletal fluorosis were investigated in each subject.Then the subjects from two villages were allocated into five subgroups(＜2.00,2.00～,3.00～,4.00～,≥5.00 mg/d),the relation fluoride intake and prevalence of osteofluorosis was analyzed.Results The prevalence of skeletal fluorosis in Wamiao Village was 31.06%(41/132),but no skeletal fluorosis case(0/35)was found in Xinhuai Village.According to the daily total fluoride intake,subjects with higher daily total fluoride intake tended to associated with a higher prevalence of skeletal fluorosis in a significant dose-response relationship(regression equation:y=2.624-6.855x+3.424x2:r=0.997).The benchmark dose lower limitation of daily total fluoride intake with 95% confidencewas 2.50 mg/d calculated according to this dose-response relationship,the reference dose(RfD)was 2.50 mg/d.In Wamiao Village a significant difference was also found between daily total fluoride intake in 41 subjects[(5.09±1.20)mg/d]with X-ray detectable skeletal fluorosis and in 91 subjects[(3.08±1.12)mg/d]without X-ray detectable skeletal fluorosis(t=-9.32,P＜0.01).Conclusions These findings indicate that the daily total fluoride intake has a significant dose-response relationship with the prevalence of skeletal fluorosis in an endemic fluorosis area associated with high-fluoride drinking water;and the RfD in this study was lower than that in the national standard of"Chinese hygienic standard for daily total fluoride intake(WS/T 87-1996)"(3.50 mg/d).

Objective To investigate the prevalence of malnutrition in human immunodeficiency virus ( HIV )‐infected children in China , and to explore and analyze the factors associated with malnutrition .Methods A cross‐sectional study was conducted by the antiretroviral treatment database of children .HIV‐infected children aged between 0 - 15 years old who initiated antiretroviral treatment were collected between January 1st , 2010 and December 31st , 2014 . Z‐score of height and weight were calculated by WHO Anthro (plus) software .Univariate and multivariate Logistic model analyses were performed to determine the factors associated with acute /chronic/mixed malnutrition .Results Baseline data of the 3 138 HIV‐infected children showed that 1 645 patients (52 .42% ) had malnutrition before antiretroviral treatment ,with acute ,chronic and mixed malnutrition of 8 .76% (275) ,39 .77% (1 248) and 3 .89% (122) ,respectively according to the type of malnutrition .Multivariate analysis showed that baseline CD4 + cell count < 200 cells/μL was the risk factor associated with acute malnutrition (aOR =2 .27 ,95% CI :1 .68 - 3 .06) ;rural settings (aOR = 1 .30 ,95% CI :1 .11 - 1 .53) ,baseline CD4 + cell count < 200 cells/μL (aOR = 1 .98 ,95% CI :1 .65 - 2 .38) ,baseline CD4 + cell count between 200 to 350 cells/μL (aOR = 1 .38 ,95% CI :1 .13 - 1 .69) and having AIDS‐related diseases (aOR = 1 .34 ,95%CI :1 .13 - 1 .59) were risk factors associated with chronic malnutrition ;and age of 11 - 15 years (aOR =2 .38 ,95% CI :1 .46 - 3 .88) ,baseline CD4 + cell count < 200 cells/μL (aOR = 4 .99 ,95% CI :3 .04 -8 .21) and having AIDS‐related diseases (aOR = 2 .45 ,95% CI :1 .65 - 3 .66) were risk factors associated with mixed malnutrition .Conclusions The prevalence of malnutrition in untreated HIV‐infected children remains high .All three types of malnutrition are associated with immunodeficiency .Early diagnosis and early treatment should be improved in HIV‐infected children through antiviral therapy to reduce the destruction of HIV to immune system .At the same time ,intensified monitoring of the nutritional status and nourishing undernourished children should be strengthened to reduce the prevalence of malnutrition .

0.05).There was significant difference in the 3-year survival rate between A and C group. Conclusions The 3-year survival rate was dramatically increased with combined therapy mainly by cisplatin, the dose of 60～80mg is tolerant for the elderly aged above seventy years, and perioperation complications can be cured.

The therapeutic effect of sustained intravitreal injectable voriconazole microspheres (VCZ-MS) on an experimental endophthalmitis of Aspergillus fumigatus was investigated. VCZ-MS was prepared successfully and its physico-chemical property was also evaluated. Right eyes of albino rabbits were infected with an intravitreal injection of 1 000 CFU x mL(-1) of susceptible Aspergillus fumigatus. All fungal endophthalmitis models were randomly divided into five groups 48 hours later: Group A is control group with no treatment; in group B, vitrectomy was performed combined with intravitreal 3 times injections of 100 microg x 0.1 mL(-1) voriconazole every other day. In group C, D and E, vitrectomy was performed combined with intravitreal injection of 0.5 mg, 1.0 mg and 1.5 mg VCZ-MS respectively. The treatment effect was assessed by slit lamp and indirect ophthalmoscope funduscopy examination, using clinical grading system of inflammation in the anterior chamber and the vitreous opacity. The optical microscopy revealed that microspheres obtained from the experiment design were opaque, discrete and spherical particles with smooth surfaces. The drug content and encapsulation efficiency of microspheres were 29.94% and 73.5%, respectively. Endophthalmitis occurred in all eyes of group A, and rapidly developed to panophthalmitis. The inflammation grade of group B, C, D or E was lower than that of group A (P < 0.05). The grade of vitreous opacity in group C, D, E is lower than group B (P < 0.05). Two eyes in group C developed to panophthalmitis. But in group D and E, all eyes whose inflammation was controlled had no recurrence with vitreous clear. Histopathological examination showed normal structures in the cured eyes, while most uncured eyes were atrophic and with eyeball destroyed. So, it can be safely concluded that the curative effect of intravitreal VCZ-MS is significantly better than that of routine intraocular injection of voriconazole. The optimal dose is the one containing 1.0 mg voriconazole.
Animals ; Antifungal Agents ; administration & dosage ; therapeutic use ; Aspergillosis ; drug therapy ; pathology ; Aspergillus fumigatus ; Delayed-Action Preparations ; Endophthalmitis ; drug therapy ; microbiology ; pathology ; Eye ; microbiology ; pathology ; Female ; Intravitreal Injections ; Lactic Acid ; Male ; Microspheres ; Polyglycolic Acid ; Pyrimidines ; administration & dosage ; therapeutic use ; Rabbits ; Random Allocation ; Triazoles ; administration & dosage ; therapeutic use ; Voriconazole

Wuzhuyu Tang is a classical formula for treating migraine, but its' pharmacological ingredients is unclear yet. Present study employed the everted intestinal sac model to collect the absorption samples of 10 kinds of Wuzhuyu decoction, and then analyzed the contents of 9 ingredients in Wuzhuyu Tang and absorption samples quantitatively or semi-quantitatively by HPLC-DAD method. Reserpine was used to establish the mice model of migraine, and then the contents and activities of 5-hydroxytryptamine, noradrenaline, dopamine, nitric oxide and nitricoxide synthase in brain tissues and serums were determined respectively after oral administration of Wuzhuyu Tang. Using the partial least squares regression method to correlate the total absorption quantity of 9 ingredients and pharmacodynamics. The result shows that limocitrin-3-O-beta-D-glucoside, ginsenoside Rg1 and Rb1, rutaevine, limonin, evodiamine and rutaecarpine are the main ingredients influenced the effects in absorption samples in everted intestinal sacs, especially ginsenoside Rg1, rutaevine, evodiamine and rutaecarpine among them have obvious improving effects to most pharmacodynamics index, might be the pharmacological ingredients influenced the therapeutical effects of Wuzhuyu Tang treating migraine.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; Male ; Mice ; Mice, Inbred ICR ; Migraine Disorders ; drug therapy ; Rats ; Rats, Wistar

Objective:To investigate the clinicopathological characteristics, treatments, and survival of patients with esophageal adeno-squamous carcinoma (EASC). Methods:A total of 494 patients with EASC were selected from the clinical information databases of 500, 000 cases with esophageal and gastric cardiac carcinomas in the Henan Key Laboratory for Esophageal Cancer Research. Among the 494 EASC cases, 361 were males with an average age of 61.47 ± 8.32 years, and 133 were females with an average age of 65.56 ± 8.06 years. SPSS 21.0 software was applied to determine the statistical differences among the different groups. A life-table method was also used to calculate the five-year survival rate. A linear regression model was used to analyze the correlation of changes at different peri-ods. Results:The incidence of EASC in our database was 0.196%(494/251707). EASC occurred predominantly in male patients (male:female=2.71:1.00). The peak age was within 60-69 years in both males and females (39.6%vs. 40.6%). Notably, the incidence of male patients showed a downward trend (R2=0.063), whereas that of female patients showed an upward trend (R2=0.004). The prevalence of EASC was obviously higher in low-incidence areas for esophageal cancer than in high-incidence areas (53.1%vs. 46.9%, P<0.001). Ac-cording to the TNM staging criteria for esophageal cancer, phases II and III patients comprised the majority of cases, which accounted for 40.8%(173/424). The positive lymph node metastasis rate was 47.0%(206/438), and the number of positive lymph node metasta-ses ranged within 1-2 (48.5%, 100/206). In addition, preoperative biopsy was performed in 467 cases, and more than half of the pa-tients (53.96%, 252/467) were diagnosed before the operation. Surgical resection was the predominant treatment method for EASC (88.8%, 419/472). Only 1.9%patients (9/472) underwent radiotherapy and chemotherapy. The five-year survival rate of male patients who were neither smoking nor drinking of alcohol was higher than that of male smokers (26.5%vs. 12.1%). In patients with stagesⅠ,Ⅱ, andⅢ+Ⅳcarcinomas with surgery as lone treatment, the three-year survival rates were 64.7%, 50.9%, and 48.5%, respectively. Correspondingly, these rates were 51.7%, 47.8%, and 33.1%after adjuvant radiotherapy and chemotherapy. Conclusion:EASC is a rare type of esophageal malignant tumor. The preoperative biopsy pathological diagnosis has high misdiagnosis rate. Smoking and drinking of alcohol can influence the prognosis of patients. In EASC patients, lymph node metastasis easily occurs, and a simple surgery is bet-ter than other cancer treatments.

Aim To explore the optimal way of breeding and genotype identification of Arrb2 knockout mice, and to find a simple and quick polymerase chain reaction ( PCR) method for the genotyping of Arrb2 knockout mice. Methods Breeding homozygote genotype of Arrb2 gene knockout mice were copula-ted with wild-type C57BL/6J mice, and then the heterozygous mating were used for mating. The growth and development of off-spring were observed. The genomic DNA was extracted from the tail of two-week-old mice. PCR was employed to amplify the Arrb2 gene fragment, and electrophoresis was used to present the gene type. Results The breeding and reproducing were successful and three genotype offspring, including wild-type,heterozygous and homozygous knockout mice were obtained. Agarose gel electrophoresis results showed the size of PCR prod-ucts was about 186 bp and 224 bp, which was consistent with the expected target gene fragment, and identified Arrb2 gene knockout mice of different genotypes successfully. Western blot analysis demonstrated the lack ofβ-arrestin2 protein in the major organs from Arrb2 -/ - mice compared with Arrb2 +/ + and Arrb2 +/ - mice. Conclusions It is feasible to obtain the homo-zygous Arrb2 knockout mice by inbreeding heterozygotes. It is simple, rapid and reliable to identify mouse genetype by PCR.