Objective To resolve the problem of the accuracy and standardization of short tandem repeat-polymerase chain reaction(STR-PCR) typing in forensic practice,the molecular clone technology has been used in producing the standard allelic ladders of DXS6799 locus.And the standard allelic ladder was applied in studying the Lisu,Pumi and De-ang populations in Yunnan Province,China.Methods Polymorphism of DXS6799 was analyzed by PCR and PAGE.Molecular cloning technology was employed to construct standard DXS6799 allelic ladder used for DXS6799 genotyping.Results The large quantities of standard allelic ladder of the locus were harvested,and the genetic polymorphisms of DXS6799 locus in three populations were studied.Conclusion The method is of high value for forensic DNA typing to construct standard ladders.DXS6799 is robust for genetic research and forensic application.

Hepatic stellate cell (HSC) activation is closely associated with the progression of liver fibrosis.As an important component of the innate immune system,natural killer (NK) cells are enriched in the liver and play a key role in host defense against viral infection and tumor,and their anti-fibrotic effect has also been confirmed.NK cells can reduce liver fibrosis by killing early-activated or senescent HSCs or secreting interferon-γ.This article summarizes related research advances in recent years,and introduces the molecular immunological mechanism of NK cells in regulating HSCs and their potential anti-fibrotic effect based on the function and phenotype of NK cells and HSCs.

Objective To investigate the genetic polymorphism of DXS8378 STR locus of chromosome X in Chinese Lisu,Pumi and De'ang populations in Yunnan and construct relative standard allelic ladders.Methods After being amplified by PCR,different STR allelic fragments were isolated from the PAG electrophoresis.The STR allelic fragments were extracted by kit and reamplified by PCR to obtain purified allelic fragments.Next,the purified allelic fragments were subcloned individually into the PUC plasmid vectors,and the size and structure of the inserts were confirmed by the analysis of their DNA sequences.Then we transfected it into competent E.coli DH5?TM cells,and finally,the recombinant plasmids DNA with the inserts were used as template for reamplification to generate the standard ladders.Results The standard allelic ladder for DXS8378 locus was obtained,with which the genetic polymorphisms of DXS8378 locus in three Chinese populations in Yunnan were studied.Conclusion The standard ladder made by this method is excellent,and DXS8378 is powerful for forensic practice in Chinese population.

ObjectiveTo study the impact of type 2 diabetes mellitus on endothelium of great saphenous vein in patients with coronary heart disease.MethodsPatients undergoing coronary artery bypass grafting were selected, 20 with type 2 diabetes mellitus (experimental group) and another 20 patients without (control group).The rings of great saphenous vein in I cm length were taken from those patients and then divided into 3 segments.The structure of endothelium was evaluated by the microscope and the changes of venous tone were measured in organ chamber at 37C with a constant supply of oxygen.Venous vasoconstriction was induced by phenylephrine (10-5 mol/L) and vasodilatation induced by nitroglycerin or acetylcholine (10-9 ～ 10-5 mol/L).ResultsMore damages of ultrastructure of the endothelium of saphenous vein were found in experimental group than in control group.There were no significant differences regarding the venous tone between the two groups (P ＞0.05) when vasoconstriction induced by phenylephrine and vasodilatation by nitroglycerin.However, the vasodilatation induced by acetylcholine was significantly decreased in experimental group than in control group (P ＜ 0.05).Conclusion Type 2 diabetes mellitus can aggravate the damage of endothelium of saphenous vein in patients with coronary artery disease.

Objective To observe the effect of smoking on the great saphenous vein tension. Methods The rings of great saphenous vein in 3mm long of selected from 31 patients with coronary artery bypass grafting were divided into three groups :smoking over 10 years( group A, n = 12 ), ex-smoking over 1 years ( group B, n =9 ) and non-smoking( group C, n =10). The changes of the tone were measured in organ chamber at 37℃ with a constant supply of oxygen when vasoconstriction induced by phenylephrine ( 10 -9 - 10 -5 mmol/L), and vasodilatation by acetylcholine ( 10 -9 - 10 -5 mmol/L) or nitroglycerin( 10 -9 - 10 -4 mmol/L)after the rings were precontracted by 10 -5 mmol/L phenylephrine. Results Vasoconstriction induced by phenylephrine and vasodilatation by nitroglycerin is no significant difference among three groups. Compared with group A, vasodilatation by acetylcholine was significantly increased in group B or C, while there is no significant difference between group B and C. Conclusion Smoking has a deleterious effect on the endothelial function of great saphenous vein, however, smoking cessation over 1 year may help to restore the endothelial function impaired by smoking.

Objective Respiratory viruses are the most common pathogens to cause respiratory tract infection in infants and children.The aim of the study was to establish a luminex-based molecular assay for rapid detection of four kinds of common respiratory viruses and provide measures for effective prevention and control . Methods 120 throat swab samples from patients with acute respiratory tract infection were collected in our hospital as disease group.30 normal specimens were used as control group .Specific up-stream and downstream primers , hybridization probes and super prim-ers were designed on the basis of conserved sequences of Influenza A and B viruses( FluA, FluB), respiratory syncytial virus types A and B ( RSVA, RSVB ) from available respiratory-virus sequence data-base.Recombinant plasmid and in vitro transcription RNA positive reference substances were established respectively .The testing sys-tem of Tem-PCR combined with luminex xMAP was built by amplification and optimization of hybridization .Comparative analysis were made between the detection results of the above method and those of single viral gene real -time PCR assay and luminex xTAG assay re-spectively. Results Rapid molecular assay was established to specifically detect the four kinds of respiratory viruses (FluA, FluB, RSVA and RSVB) with the sensitivity of 10 copies/μL.Rapid molecular assay and single viral real-time PCR assay were utilized to de-tect the throat swabs ( n=120 ) from suspected patients , the positive result of the former was 31 .7% ( 38/120 ) and the latter was 29.2%(35/120).The consistency test result indicated the two methods were consistent without a significant difference (k>0.7). Several samples were detected by luminex xTAG assay simultaneously , in which good consistency and significant difference were found in two assays by statistical analysis (k>0.6). Conclusion Preliminary clinical application has confirmed the novel molecular assay is sensitive, specific and rapid in simultaneous detection of FluA , FluB, RSVA and RSVB respiratory viruses , which provides experi-mental basis for accurate diagnosis of infected pathogens at early clinical stage .

ObjectiveTo estimate the application of mycophenolate mofetil (MMF) and cyclophosphamide(CTX) intravenous pulse therapy on diffuse proliferative lupus nephritis (DPLN).MethodPubMed,Medline,EMBASE and CNKI were searched from the establishment of the database.Meta-analysis of 14 comparative studies on MMF and CTX in treatment of DPLN was performed,taking the remission,the relapse,the death of MMF and CTX for DPLN as primary efficacy variable,mean while taking the herpes zoster as safety evaluating indicator.ResultsMMF was better than CTX in remission rate ( P ＜ 0.05 ).There was no difference between in incidence rate of the relapse,the death and the herpes zoste MMF and CTX for DPLN ( P ＞ 0.05 ).ConclusionMMF was better than CTX on the efficacy and safety in DPLN.

The development of health-related quality of life (HRQL) instrument in traditional Chinese medicine (TCM) is increasing rapidly in China, but few studies focus on their necessity. This study explores the necessity of the development of TCM instruments from both theoretical and practice perspectives, and aims to explain whether the adoption of the cross-medical style is valid. Through theoretical analysis, both TCM and Western medicine instruments show the same objectives, whereas TCM instruments are more suitable for the Chinese social behavior, customs and expectations. In practical analysis, 47 TCM instruments were identified, among which 17 had 18 corresponding Western medical instruments. In the domains layer, except for physiological, psychological and social factors, TCM instruments focus more on the harmony between body and spirit, humanity and nature or human and society and the constitution, etc. In the facts layer, TCM instruments focus on the emotions, initiative social intercourse, TCM symptoms, diet, sleep, taste, feces and urine, etc. In addition, significant differences existed in the methods of information selection. There is no need to modify cross-medical style research except when TCM characteristic terms exist, but attention must be paid to the influence of culture in different areas. Therefore, the TCM instruments can resolve the limitations of the application of Western medical instruments to the Chinese setting, while also having remarkable abilities of information coverage and detection. Both forms of instruments have the capacity and requirement to inter-communicate with each other in order to serve the whole Chinese cultural system. Generally speaking, there is no need to modify the instruments in cross-medical style research. But this point requires further demonstration in the rigorous designed clinical trials.

Objective: To explore the expression of the A_2A adenosine receptor and the inflammatory cytokines interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) in human degenerative nucleus pulposus (NP) cells after they have been treated with a pulsed electromagnetic field (PEMF). Methods: Human degenerative NP cells were cultured in vitro and treated using an 0.8mT PEMF with a pulse frequency of 50Hz. The pulse width was 150μs and the exposure time was 30min, repeated 5 times at 12 hour intervals. The expression of the A_2A adenosine receptor in NP cells was determined using western blotting and reverse transcription polymerase chain reactions. The expression of the inflammatory cytokines IL-1β and TNF-α were detected using enzyme-linked immunosorbent assays (ELISA). The human degenerative NP cells were also treated with an antagonist and agonist of the A_2A adenosine receptor, and the expression of IL-1β and TNF-α were also determined using ELISA. Results: After the PEMF treatment the expression of the A_2A adenosine receptor increased significantly, while the expression of IL-1β and TNF-α decreased significantly. However, the A_2A adenosine receptor antagonist reversed the inhibitory effect of the PEMF on the expression of IL-1β and TNF-α, while the agonist played an opposite role. Conclusion A PEMF can significantly inhibit the expression of IL-1β and TNF-α in human degenerative NP cells, which could be related to up-regulation of the expression of the A_2A adenosine receptor in those cells.

Chronic hepatitis B induced by HBV infection is a significant risk factor leading to liver cirrhosis and liver cancer. Half a century ago， HBeAg was first discovered in the serum of HBV infected individuals， and although HBeAg does not participate in HBV infection or replication in hepatocytes， studies have shown that it can interfere with the innate and adaptive immune responses of the host and play an important role in immune activation and immunosuppression during chronic HBV infection. HBV has no cytotoxicity to the infected hepatocytes， and the antiviral action and inflammatory response mediated by immune response determine whether HBV is cleared or induces liver inflammation-related diseases. Therefore， this article reviews the formation of HBeAg and its immune activation and immunosuppression mechanisms in chronic HBV infection， with a focus on the different immune effects caused by HBeAg on innate immune and adaptive immune cells， and this article also elaborates on the dual role of HBeAg in inducing immune responses and explores the conversion role of HBeAg in different stages of chronic HBV infection.