BACKGROUND:Kalikrein 1 is an important component of the kalikrein-kinin system. Studies have shown that kalikrein can protect the cardiovascular system by promoting angiogenesis and inhibiting myocardial inflammation, but there is no report on its effect on inducing differentiation of stem cels. OBJECTIVE: To determine the transfection efficiency of kalikrein 1 adenoviral vector in rat bone mesenchymal stem cels. METHODS:Using adenovirus as a vector, the target gene kalikrein 1 was transfected into rat bone marrow mesenchymal stem cels. Fluorescence microscopy, MTT method and flow cytometry were used to investigate the effect of transfection and determine the optimal multiplicity of infection. RESULTS AND CONCLUSION:Adenovirus carrying kalikrein 1 was successfuly transfected into rat bone marrow mesenchymal stem cels. Results from flow cytometry showed that the transfection efficiency was associated with the multiplicity of infection. When the multiplicity of infection was 150, the transfection efficiency was 80.8%. MTT results showed that when the multiplicity of infection was 200, the cel growth was inhibited remarkably. These findings indicate that adenovirus-mediated kalikrein 1 can be successfuly transfected into rat bone marrow mesenchymal stem cels with the optimal multiplicity of infection=150.

Objective To establish and evaluate a universal real-time fluorescent quantitative PCR(qPCR)method for identifying and quantifying three human parvovirus B 19 ( B19V) genotypes.Methods Firstly, following a bioinformatic analysis of a subset of B19V genomic sequences available in the NCBI nucleotide database ,representative of genotypes 1 to 3,a set of suitable universal primers and TaqMan probes was designed from the NS 1 gene of B19V.Aplasmid was used as a quantitative standard that contained the identical sequence of the B 19 target sequence .An internal control ( IC ) was included to prevent false negative results .Then,serial 1-log dilutions of quantitative standards were prepared and used in the qPCR assays for generation of a standard curve .Finally,the specificity,sensitivity and reproducibility of the assay were assessed.Results A linear relationship of the real-time PCR method for detecting B19V from 1 ×109copies/μl to 1 ×103 copies/μl was observed .The developed qPCR protocols allowed for the detection of genotypes 1 to 3 with a limit of detection ( LOD) of 10 copies/μl.Furthermore, the assay did not amplify other blood-borne viruses.The inter-and intra-assay variability analyses showed good reproducibility of the assay .Conclusion A universal real-time qPCR method for the detection of B19V DNA is established,which will facilitate the diagnosis of B19V infections and the screening of blood and plasma-derived products , thereby improving the viral safety of transfusion and plasma-derived products .

Randomized controlled trials (RCT) is the source of the raw data of evidence-based medicine. Blind method is adopted in most of the high-quality RCT. Sham acupuncture is the main form of blinded in acupuncture clinical trial. In order to improve the quality of acupuncture clinical trail, based on the necessity of sham acupuncture in clinical research, the current situation as well as the existing problems of sham acupuncture, suggestions were put forward from the aspects of new way and new designation method which can be adopted as reference, and factors which have to be considered during the process of implementing. Various subjective and objective factors involving in the process of trial should be considered, and used of the current international standards, try to be quantification, and carry out strict quality monitoring.
Acupuncture Therapy ; standards ; Biomedical Research ; standards ; Evidence-Based Medicine ; Humans ; Randomized Controlled Trials as Topic ; standards

Objective To observe the changes of the peripheral blood T helper (Th)17 cells/regulatory T cells (Treg) ratio in patients with anti‐tuberculosis drug‐induced liver injury ,and to explore the relationship between Th17/Treg ratio and liver function . Methods Ninety inpatients with the diagnosis of secondary pulmonary tuberculosis (TB) from Affiliated Hospital of Zunyi Medical College from Feb .2013 to Mar .2015 were included in this study .All these patients had normal base line liver function .After 2 weeks of anti‐TB chemotherapy ,21 patients with alanine transaminase (ALT) and/or aspartate aminotransferase (AST) ≥2 × upper limit of normal (ULN) were defined as TB with liver injure group ,the other 69 patients with ALT and/or AST<2 × ULN were defined as TB with non‐liver injured group .Before treatment and two weeks after anti‐TB therapy ,the peripheral blood Th17/Treg ratios and liver functions of all patients in both groups were detected in order to compare the changes of Th17/Treg ratios pre and post anti‐TB chemotherapy and to explore the relationship between Th17/Treg ratio and ALT/AST levels .Student′s t test was used for statistical analysis ,and Pearson correlation analysis was used for correlation test .Results Before treatment ,there was no statistical significance of Th17 ,Treg and Th17/Treg ratio between groups (P> 0 .05) .The Th17 levels in TB with liver injured group ([2 .522 ± 0 .388]/mL vs [2 .075 ± 0 .369]/mL ,t=3 .633 ,P<0 .01) and TB with non‐liver injured group ([2 .326 ± 0 .348]/mL vs [1 .929 ± 0 .402]/mL ,t= 6 .468 , P< 0 .01) ,and Th17/Treg ratios in both groups(0.618±0.104vs0.489±0.107,t=3.553,P<0.01 ;0.554±0.108vs0.450±0.098,t=6 .353 ,P<0 .01) were all increased after anti‐TB chemotherapy ,with statistically significant differences . However ,there was no statistical significance of Treg levels pre and post anti‐treatment in both groups (both P>0 .05) .The Th17 level and Th17/Treg ratio in TB with liver injured group were significantly higher than those in TB with non‐liver injured group (t= 2 .203 , P< 0 .05 and t= 2 .345 , P< 0 .05 , respectively) .In liver injured group ,the Th17/Treg ratio was positively correlated with ALT level (r=0 .849 ,P=0 .044) ,but not with AST level (P>0 .05) .Conclusion The changes of Th17/Treg ratio are related with anti‐TB drug‐induced liver injury in patients with secondary pulmonary tuberculosis .

Abstract: Schistosomiasis, an important zoonotic parasitic disease, is one of the six major tropical diseases identified by WHO, and also one of the most important parasitic diseases for prevention and control in China. After more than 70 years of efforts, the prevention and control of schistosomiasis in China has made great achievements, and the current epidemic of schistosomiasis in China has entered an extremely low epidemic state, but the distribution base of the only intermediate host of schistosomiasis, Oncomelania hupensis, is still large. For now, the techniques used to monitor schistosomiasis have shortcomings such as time-consuming, laborious and low sensitivity, which cannot meet the current needs of China. Environmental DNA (eDNA) refers to DNA that can be extracted from environmental samples (such as soil, water or air) without isolating any target organisms, which is a complex mixture of genomic DNA and its degradation products from different organisms in the same environment. eDNA technology can reflect the community or species composition information in the ecosystem through DNA extraction and detection of environmental samples. Compared with traditional biological monitoring methods, eDNA technology has the advantages of high efficiency, high sensitivity and environmental friendliness. eDNA has been successfully used for the specific detection of Schistosoma mansoni, Schistosoma haematobium and Schistosoma japonicum. This paper reviews the current detection methods of eDNA, the application and technical limitations of eDNA technology in schistosomiasis monitoring, aiming to provide scientific reference for research in the field of schistosomiasis surveillance.

Objective To detect human parvovirus B19(B19V)DNA in source plasma pools and coagulation factor products and determine its prevalence and the level of contamination .Methods A pair of primers and a probe selected from the highly conserved sequences encoding the non-structural protein(NS1)of B19 were designed and synthesized.With the primer-probe combination ,source plasma pools and four types of coagulation factor products were determined for B 19V DNA by TaqMan real-time quantitative PCR.Results One-hundred and sixteen from 195 (59.49%) source plasma pools contained B19 DNA and concentrations up to 1.35 ×1010 copies/ml were measured.High frequencies of contamination were detected in factor Ⅷ (29 of 31; 93.55%), thrombin (10 of 10; 100%), fibrinogen (6 of 7; 85.71%) and prothrombin complex (8 of 9;88.89%).Conclusion These data show that B19V is a common contaminator in Chinese source plasma pools and coagulation factor products .Thus,B19V screening in Chinese source plasma seems desirable and significant for the safety of plasma derivatives in China .

Objective To study the imaging findings of disseminated pulmonary tuberculosis in patients with acquired immunodeficiency syndrome (AIDS).Methods X-ray and multi-slice CT (MSCT)data from 33 AIDS patients with disseminated pulnonary tuberculosis confirmed by clinical manifestations and laboratory tests were analyzed retrospectively.Results Thirty patients underwent initial chest radiography examination,29 patients showed abnormal appearances,including bilateral disseminations in 21 patients and unilateral multiple disseminations in 8 patients.All patients underwent MSCT examination,26 patients showed bilateral disseminations and 7 patients showed unilateral multiple disseminations.The abnormal pulmonary appearances included nodule (n =25),miliary nodule (n =22),air-space consolidation (n =22),cavity (n =11),fibrosis (n =7),ground-glass opacity (n =7),pneumatocele (n =4),calcification (n =2).There were 20 patients with more than 3 abnormal appearances and 13 patients with one or two abnormal appearances.The extra-pulmonary tuberculosis included pleural effusion (n =33),lymphadenopathy (n =30),intestinal tuberculosis (n =3),splenic tuberculosis (n =1) and cerebral tuberculosis (n =1).Conclusion Disseminated pulmonary tuberculosis should be highly suspected in AIDS patients with diffused nodules,miliary nodules,air-space consolidations or multiple cavities,accompanied with pleural effusion and lymphadenopathy.

Objective To evaluate the clinical efficacy of the intratympanic inflation by tympanum inflatable balloon for patients with Meniere’s disease(MD) .Methods A total of 36 confirmed cases (according to the listening stage) of Ⅱ - Ⅲ period of MD patients were randomly divided into two groups :experimental group and control group ,18 cases of matched group ,who were staged before preoperation by hearing ,Ⅱ period (average threshold 25~40 dB HL) 6 cases ,Ⅲ period (41~70 dB HL) 12 cases ,were treated by intratympanic methylprednisolone per‐fusion after putting tympanostomy tube under endotoscope ,then once every 3 days perfusion with a total of three times .There were 18 cases of experimental group staged before preoperation by hearing ,Ⅱ period (average thresh‐old 25~40 dB HL) in 5 cases ,Ⅲ period (41~70 dB HL) 13 cases treated by using tympanum inflatable balloon in the tympanum under endotoscope .Two groups were given conventional improved microcirculation and nutritional therapy .Results In 18 cases of patients in experimental group ,for the 18 to 24 months follow -up after treatment ,6 cases of vertigo were fully controlled (grade A) ,10 cases of vertigo basically controlled (grade B) ,2 cases of vertigo partly controlled (grade C) .Vertigo effective control rate was 88 .89% (16/18) .After 18 to 24 months treatment ,two groups of 36 patients with tinnitus and pure tone average threshold had no obvious change .In 18 ca‐ses of patients in the matched group ,during the 18 to 24 months follow -up after treatment ,3 cases of vertigo were fully controlled (grade A) ,7 cases of vertigo basically controlled (grade B) ,8 cases of vertigo partly controlled (grade C) .Vertigo effective control rate was 55 .56% (10/18) .The experimental vertigo basic control rates were higher than the control group (χ2 = 4 .985 ,P<0 .05) .The activitives in the two groups had improved significantly . No obvious differences were found between two groups .Conclusion With the intratympanic gasing by tympanum in‐flatable balloon for patients of Ⅱ - Ⅲ Menier’s disease ,control rates was high and motile ability were improved , but tinnitus and hearing improvement was not obvious .

Langerhans cell tumor is a kind of tumor that originates from Langerhans cells (LC) and maintain their specific phenotype profile and ultrastructural features. Based on cell morphology, immunohistochemical and ultrastructural characteristics, Langerhans cell tumor has two main subcategories: Langerhans cell histiocytosis (LCH) and Langerhans cell sarcoma (LCS). LCH is a benign clonal proliferative disease of the LC, whereas LCS is an extremely rare neoplastic proliferation of Langerhans cells with overtly malignant cytologic features and spreads aggressively, which is considered to be a high level malignant type of LCH. Both LCH and LCS can involved various tissues and organs and have complex and diverse clinical manifestation, which cause different severity. The diagnosis depends on histopathological morphology and immunohistochemistry; the electron microscopy was used to assists diagnosis when necessary. The treatment includes surgery, chemotherapy, radiotherapy, immunotherapy and hematopoietic stem cell transplantation, etc, but lack of generally accepted optimal treatment options currently, individualized treatment is needed. The prognosis of LCH is primarily related to the number of damaged organ, while LCS has a poor overall prognosis as its invasion and rapid progress. This article reviews the pathogenesis, clinical manifestations, diagnosis, treatment and prognosis of both LCH and LCS.
Histiocytosis, Langerhans-Cell ; Humans

Abstract A method based on dispersive solid phase extraction-high performance liquid chromatography-tandem mass spectrometry for determination of 19 kinds of nonprotein nitrogen compounds including melamine, cyromazine, amidinourea, aminotriazine, 3-aminotriazole, 4-aminotriazole, allantoin, cyanuric acid, dicyandiamide, thiourea, semicarbazide, L-leucine, L-isoleucine, L-arginine, L-hydroxyproline, L-theanine, ammeline, ammelide and guanidine in powdered formulas was presented. The nonprotein nitrogen compounds were degreased by chloroform and extracted by acetonitrile, with MgSO4 to remove water and C18 to clean up. The samples were separated on Merck ZIC HILIC column (150 mm í2. 1 mm, 5 μm, 20 nm) with gradient elution. The electrospray ionization was operated in the positive mode and negative mode, and monitored by the multiple reaction monitoring ( MRM) mode. Allation was quantified by external standard method and the other 18 kinds of nonprotein nitrogen compounds were quantified by internal standard method. All of the correlation coefficients (r) were higher than 0. 99. The limits of quantitation (LOQ) were 0. 05-5. 0 mg/kg, the average recoveries were between 82 . 2% and 115 . 0%, and the relative standard deviations were less than 20%.