This report concerns an observation on the biological properties of orthomyxoviruses type A in the condition of symbiosis with streptococcus A in embryonated eggs. The results show that both the viruses and the streptococci multiply normally in the condition of symbiosis and that the activity of the virus neuraminidase enhances, with the speed of the virus disassociating from RBC increasing. The morphological changes of the virus were observed also by transmission electron microscopy. The reason of these changes is discussed,

A method for determinattion of monoamine neurotransmitters and related compounds in microdialysis solution by high performance liquid chromatography (HPLC) with pulsed amperometric detection was described. A comparison of response of iso potential anperometric (AD) and pulsed amperometric detection (PAD) was made. The results indicated that the sensitivity of PAD was 1.2 fold that of AD. The quantitative determination of monoamine neurotransmitters and related compounds was carried out with 3,4-dihydroxybenzylamine (DHBA) as internal standerd. The recoveries of monoamine neurotransmitters in microdialysis solution are 71% ～ 101%.

Objective To compare the clinical efficacy of locking plate (LCP),dynamic hip screw (DHS)and proximal femo-ral nail (PFNA)internal fixation in the treatment of elderly stable femoral intertrochanteric fracture.Methods A total of 60 pa-tients with stable stable intertrochanteric fractures in elderly patients were randomly divided randomly into proximal femoral LCP fixation group (group A),the DHS fixation group (group B)and PFNA fixation group (group C).The operative time,blood loss, postoperative complications and postoperative weight-bearing time were analyzed among three different groups.Results There were significant difference in blood loss,postoperative weight-bearing time,operative time among three different groups (P < 0.05 ). Group C was significantly better than the group A(P <0.05).Two cases of hip varus deformity,one case of crew loosening and su-perficial infection were appeared among group A.One case of hip varus deformity occurred in group B.One case of suffered refrac-ture occurred in group C.Conclusion LCD fixation,fixation with DHS and PFNA fixation are effective treatment for stable femoral intertrochanteric fracture in the elderly patients.

Objective To establish the crystal violet plaque assay for detection of virus titer of recombined Tiantan vaccinia AIDS vaccine,and provide more stable method of virus titration for rTV AIDS Vaccine.Methods Optimized the concentration of Vero cells,the time and temperature of virus adsorption,and the time of determination for CPE,then established the crystal violet plaque assay for virus titer of rTV.Counting and analysis the plaques by BioSpot Reader,then analyzed the relativity of plaques counted with BioSpot Reader and manual; Several lots rTV AIDS Vaccine and Tiantan vaccinia were titrated by the method of plaque formation-hemadsorption assay,neutral red and crystal violet plaque assay,then analyzed the relativity of the results of three methods ; meanwhile,the virus titer of samples were determine repeatedly by the crystal violet plaque assay,then calculated the coefficient of variation( CV),and verified the precision of the method; SPSS17.0 was used in statistical analysis of the experimental results.Results When the concentration of Vero cells was 5.0×105-9.O×105 cells/ml,virus been adsorbesd 2 h at 37℃,then cultivated 72 h after adding the culture medium containing methyl cellulose.Plaques counted by BioSpot Reader was highly related with counted by manual (r =0.985),so BioSpot Reader counting can objectively reflect the virus plaques with various size,and reduce the error by manual counting; compared the virus titration for different lots of rTV AIDS vaccine and Tiantan vaccinia with three methods,the crystal violet plaque assay was highly related with plaque formation-hemadsorption assay (r =0.997,P＜0.01 ) and neutral red plaque assay(r=0.980,P＜0.01 ).Conclusion Crystal violet plaque assay was established for virus titration of rTV AIDS Vaccine.

Objective To study the genetic diversity of Fritillaria thunbergii,a traditional Chinese herb in Zhejiang Province in China.Methods The genetic diversity of six representational populations of F.thunbergii including 32 individuals was investigated by amplified fragment length polymorphism(AFLP) maker technique.Results The genetic diversity was revealed as follow: the Nei′s genetic diversity index(He) 0.169 0?0.175 7,Shannon′s information index(I) 0.269 8?0.245 3,percentage of polymorphic loci(PPB) was 76.85% at the species level;Ht 0.169 0?0.030 9,and Hs 0.150 8?0.024 0,I 0.233 3?0.261 9, PPB was 50.38% at population level.The genetic differentiation index(Gst) was 0.107 6,Nm 4.147 0.The result of dendrogram of six populations indicated that Dongyang and Yongkang populations shared the minimum genetic distance(0.015 0),they were classified into a group,and Xiangshan and Jinyun populations shared the maximum genetic distance(0.032 4).Conclusion The genetic diversity of F.thunbergii cultivated in Zhejiang Province is very rich,which could ensure the long-term survival of F.thunbergii.But the genetic diversity of F.thunbergii is relatively higher in population levels while lower at the species levels and the degree of genetic differentiation occured among the populations is not significant.The germplasm resources are relatively stable among these six populations.These populations could be used to breed the fine strains of F.thunbergii as the bases.

BACKGROUND: LIM Mineralization protein 1 (LMP-1), an intracellular non-secretory protein, plays roles in bone calcification. Presently, it is found that LMP-1 can promote an increase in bone morphogenetic protein 2 and transforming growth factor ? 1. This indicates that LMP-1 may recruit a mass of ossified factors to participate in the differentiation of osteoblasts. OBJECTIVE: To construct human LMP-1 gene adenovirus recombinant with AdEasy adenovirus vector system, and to detect LMP-1 expression in infected rabbit bone marrow mesenchymal stem cells (BMSCs). DESIGN, TIME AND SETTING: An opening experiment was performed at the Central Laboratory of South West Hospital, Third Military Medical University of Chinese PLA from March 2006 to February 2007. MATERIALS: Three New Zealand rabbits were used to isolate bone marrow mesenchymal stem cells. Plasmid pIRES2-EGFP-LMP-1 carrying human LMP-1 gene was kept in Department of Orthopedics, Second Hospital Affiliated to Chongqing Medical University. AdEasy was presented by Dr. Tong-Chuan He from USA. Human embryo kidney 293 cells were gifted by Wang from Department of Clinical Laboratory of Chongqing Medical University. METHODS: LMP-1 gene with a sequence encoding His-tag was amplified by using pIRES2-EGFP-LMP-1 plasmid as a template for polymerase chain reaction (PCR) with a specially designed downstream primer. The target gene was cloned to the pMD18-T vector for sequencing. Once verified, the gene was cut out by double endonucleases, connected to the shuttle vector pAdTrack-CMV. The newly constructed vector was linearized by PmeⅠ following efficient homologous recombination with the backbone vector pAdEasy-1 in BJ5183. The correct recombinant pAd-LMP-1 was linearized with Pac I and transfected to HEK293 cell by means of mediated Lipofectamine. The titer of virus was measured after amplification and purification. The mRNA and protein expression of LMP-1 was detected in BMSCs, which were infected with Ad-LMP-1 at the most appropriate MOI, were detected by reverse transcription (RT)-PCR and Western blot, respectively. MAIN OUTCOME MEASURES: Plasmid pAd-LMP-1 identification, its titre and efficiency of infection. mRNA and protein expression of LMP-1 were detected by RT-PCR and Western blot, respectively. RESULTS: The recombinant plasmid pMD18-T-LMP-1 carrying LMP-1 gene with His-tag was successfully constructed. After packaging and amplification of the recombinant adenovirus, the 3.5?109 efu/ml titer of Ad-LMP-1 was obtained by CsCl gradient purification. The optimal efficiency infection was 50%-70%, which was get after Ad-LMP-1 infected BMSCs for 3 days at the most appropriate MOI 150. The mRNA and protein expression of LMP-1 in infected BMSCs had been proved. CONCLUSION: The recombinant adenovirus containing human LMP-1 gene with His-tag is successfully constructed. The BMSCs infected with recombinant adenovirus Ad-LMP-1 can effectively express LMP-1.

Objective To systematically assess the detection of K-ras gene mutation at codon 12 in pancreatic juice for diagnosis of pancreatic cancer.Methods A comprehensive electronic searching was performed to retrieve relevant studies on K-ras gene mutation in pancreatic juice for diagnosis of pancreatic cancer.Data on accuracy of included studies were extracted,Meta-DiSc1.4,Stata10.0,was applied for further heterogeneity exploring,meta-analysis and publication bias testing.Results Fifteen studies met the inclusion criteria.Heterogeneity was not found among these studies,including threshold effect,different test methods,randomization and blind.Pooled accuracy indicators like sensitivity,specificity and diagnostic odds ratio(DOR) were 0.61(95%CI 0.56-0.66),0.82(95%CI 0.77-0.86) and 6.28(95%CI 4.42-8.91),respectively.Area under curve(AUC) of SROC(summary receiver operating characteristics) was 0.8207 and Q index was 0.7542.Publication bias had little influence on meta-analysis with the fail-safe number of 694.0108.Conclusions This meta-analysis demonstrates that the detection of K-ras gene mutation in pancreatic juice has moderate effect for diagnosis of pancreatic cancer.It is not enough to be an early diagnosing or screening indicator for pancreatic cancer,but it can be used clinically as an important adjunct to cytology,imageology and enzymology in the diagnosis of pancreatic cancer.

This article was aimed to explore the effect ofSan-Huang (SH) decoction on improving chemosensitivity of MCF-7 cells to epirubicin and inhibition of Aurora kinase A, in order to discuss its underlying mechanism. The inhibition of MCF-7 cells proliferation on breast cancer by SH decoction was determined by CCK-8 assay. RT-PCR and western blot were used to detect the Aurora A, p53 mRNA and protein expression level of MCF-7 cells by SH decoction. The siRNA silenced Aurora A of MCF-7 cells. CCK-8 assay was used to detect the inhibition of MCF-7 cells proliferation. CCK-8 assay and AnnexinV-FITC/PI staining were used to detect the inhibition rate and apoptosis rate of MCF-7 cells treated by the combination of SH decoction and epirubicin. Western blot analysis was used to detect the expression of apoptosis-related proteins. The results showed that SH decoction inhibited the proliferation of MCF-7 cells in a dose-dependent manner (P< 0.05). The effect of 48 h medication was better than 24 h (P < 0.05). There was no statistical difference with medication for 72 h (P > 0.05). SH decoction can regulate the Aurora A, p53 protein and mRNA expression of MCF-7 cells. siRNA silenced Aurora A, which downregulated the inhibition rate of MCF-7 cells by SH decoction for 50.0% (from 49.2% to 24.8%). The combination of SH decoction and epirubicin enhanced the effect of epirubicin on inhibiting the proliferation rate and apoptosis rate of MCF-7 cell, regulated the expression levels of apoptosis-related protein such as c-PARP, c-Caspase 3, Bcl-2, Bax, as well as the protein level of Aurora A. It was concluded that SH decoction can increase the chemosensitivity of MCF-7 cells to epirubicin, which may be related to the inhibition of Aurora Kinase A by SH decoction.

Objective:To explore the risk factors of thyroid nodules in diabetic patients and its correlation with Traditional Chinese Medicine (TCM) constitution.Methods:A Total of 213 cases of diabetic patients in Guang’anmen Hospital and Tangshan Hospital from January 2019 to August 2020 were choosen to do the questionnaire, with containly symptom and constitution. The patients were divided into diabetes with thyroid nodules group and diabetes without thyroid nodules group according to whether thyroid nodules were combined. We compared the clinical data characteristics of 2 groups, and used multi-factor logistic regression model to analyze the risk factors of diabetic patients with thyroid nodules and their correlation with TCM constitutions. Results:Diabetes patients aged from 50-80 years old [ OR=2.949, 95% CI (1.266-6.714)], females [ OR=3.736, 95% CI (1.823-1.541)], diabetes duration≥15 years [ OR=1.558, 95% CI (1.623-1.585)], elevated HbA1c [ OR=5.862, 95% CI (1.418-23.629)], elevated VLDL [ OR=2.851, 95% CI (1.597-6.824)], frequent insomnia [ OR=1.970, 95% CI (1.315-3.395)], Qi stagnation [ OR=4.357, 95% CI (2.634-8.377)], blood stasis [ OR=4.420, 95% CI (1.874-15.258)] are more likely to suffer from thyroid nodules ( P<0.05). Conclusion:Diabetic patients aged from 50-80 years old, females, diabetes duration≥15 years, elevated HbA1c, family history of thyroid nodules, frequent insomnia, and mood swings are more likely to develop thyroid nodules; qi stagnation and blood stasis are dangerous constitutions for diabetic patients with thyroid nodules.

Objective:To observe and preliminary investigate the influencing factors of exudative retinal detachment in patients with severe preeclampsia (S-PE) and its influence on pregnancy outcome.Methods:A total of 89 patients of S-PE patients diagnosed in Department of Obstetrics in The Third Central Hospital of Tianjin from July 2018 to June 2020 were included in this study. According to whether the patients were complicated with exudative retinal detachment, they were divided into retinal detachment group (43 cases) and non retinal detachment group (46 cases). All patients underwent direct ophthalmoscopy, and those with suspected or definite retinal detachment underwent optical coherence tomography or B-mode ultrasonography. The age, course of disease, body mass index, systolic blood pressure, diastolic blood pressure, laboratory serum albumin level, 24 h urinary protein, platelet count, mean platelet volume, platelet distribution width and serum uric acid content were collected. The average age ( t=-1.01), body mass index ( t=-1.799), course of disease ( t=-1.657) and constituent ratio of primipara and multipara in the two groups were compared ( χ2=1.411), the difference was not statistically significant ( P>0.05). The end of pregnancy time, systemic complications, fetal outcomes and neonatal Apgar scores were analyzed, retrospectively. Independent sample t test was used to compare the quantitative data between the two groups; Chi square test was used to compare qualitative data. Binary logistic regression analysis to screen the influencing factors of exudative retinal detachment in S-PE. Results:Systolic blood pressure and diastolic blood pressure were higher in the retinal detachment group than in the non retinal detachment group, and the difference was statistically significant ( t=2.087, 2.810; P=0.040, 0.006). Serum albumin ( t=-10.404), 24-hour urinary protein quantification ( t=6.401), blood uric acid ( t=3.910), platelet count ( t=-6.124) and platelet distribution width ( t=2.992) were compared between the two groups, and the differences were statistically significant ( P<0.05). There were 30 (69.77%, 30/43) cases in retinal detachment group and 14 (30.43%, 14/46) cases in non retinal detachment group with severe systemic complications; The average time of ending pregnancy was 34.66±3.29 and 38.25±1.62 weeks; The average Apgar scores at birth were 7.28±3.40 and 9.46±1.59. The incidence of severe systemic complications in two groups ( χ2=13.755), the end of pregnancy time ( t=-6.448), Apgar score at birth ( t=-3.824), the differences were statistically significant ( P=0.000、0.000、0.000). Fetal distress and death were 8 (18.60%, 8/43), 7 (16.28%, 7/43) and 7 (15.22%, 7/46), 1 (2.17%, 1/46) in retinal detachment group and non retinal detachment group respectively. There was no significant difference in the incidence of fetal distress between the two groups ( χ2=0.182, P=0.670). The difference of fetal mortality was statistically significant ( P= 0.027). Logistic regression analysis showed that serum albumin and uric acid were associated with exudative retinal detachment in patients with S-PE ( P<0.05). Conclusions:Serum albumin and uric acid are the influencing factors of exudative retinal detachment in patients with S-PE. Patients with exudative retinal detachment are prone to adverse pregnancy outcomes.