Detection of PERV by polymerase chain reaction and its safety in bioartificial liver support system

瞄准：为了建立在中国试验性的微型猪检测猪的内长的制动火箭病毒(PERV ) 的一个方法并且在三评估 PERV 的安全，个人们基于猪的 hepatocytes 与简历 artificial 肝支持系统对待。方法：猪的 hepatocytes 与二阶段的灌注方法被孤立，在生物反应器然后有教养，它被半渗透的膜(0.2 microm ) 与病人的血浆通过被传播的腔分开。在 post-hemoperfusion 以后，病人的血为屏蔽被获得。另外，媒介的样品从实验室生物反应器的管腔内、额外的钠分隔空间收集了，文化上层清液在试管内被分析。病毒的序列的存在被聚合酶链反应(PCR ) 和反向的 transcriptase 聚合酶链反应(RT-PCR ) 估计。最后，在普通文化的上层清液的病毒的感染被暴露查明到胎儿的肝细胞。结果：PERV 特定作呕序列用 RT-PCR 在猪的 hepatocytes 被发现。并且从管腔内的、额外的钠样品和文化上层清液在所有样品被检测。然而，从主要猪的 hepatocytes 的文化上层清液(细胞的碎片变清) 没能感染人的胎儿的肝细胞。最后， RT-PCR 没检测感染在在各种各样的时间 post-hemoperfusion 从三个病人获得的血样品被发现的 PERV。结论：使用的试金特定、敏感，由第二 PCR 识别了。没有 mitogen 的刺激， PERV 能被免除在生物反应器有教养的 hepatocytes 并且不能被空纤维阻止半渗透的膜，显示在身体外的简历 artificial 肝的 PERV 安全的存在支持系统(EBLSS ) 。

Screening of genes of proteins interacting with p7 protein of hepatitis C virus from human liver cDNA library by yeast two-hybrid system

AIM: To investigate the biological function of p7 protein and to look for proteins interacting with p7 protein in hepatocytes.METHODS: We constructed p7 protein bait plasmid by doning the gene of p7 protein into pGBKT7, then transformed it into yeast AH109 (a type). The transformed yeast was mated with yeast Y187 (α type) containing liver cDNA library plasmid, pACT2 in 2xYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-α-gal for selection and screening. After extracting and sequencing of plasmids from blue colonies, we performed sequence analysis by bioinformatics.RESULTS: Fifty colonies were selected and sequenced.Among them, one colony was Homo sapiens signal sequence receptor, seven colonies were Homo sapiens H19, seven colonies were immunoglobulin superfamily containing leucine-rich repeat, three colonies were spermatid peri-nuclear RNA binding proteins, two colonies were membrane-spanning 4-domains, 24 colonies were cancer-associated antigens, four colonies were nucleoporin 214 ku and two colonies were CLL-associated antigens.CONCLUSION: The successful cloning of gene of protein interacting with p7 protein paves a way for the study of the physiological function of p7 protein and its associated protein.

Screening of hepatocyte proteins binding to F protein of hepatitis C virus by yeast two-hybrid system

AIM: To investigate the biological function of F protein by yeast two-hybrid system.METHODS: We constructed F protein bait plasmid by cloning the gene of F protein into pGBKT7, then recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-HisAde) containing X-α-gal for selection and screening.After extracting and sequencing plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics.RESULTS: Thirty-six colonies were selected and sequenced.Among them, 11 colonies were zymogen granule protein,5 colonies were zinc finger protein, 4 colonies were zinc-α-2-glycoprotein, 1 colony was sialyltransferase, 1 colony was complement control protein factor Ⅰ, 1 colony was vitronectin, and 2 colonies were new genes with unknown function.CONCLUSION: The yeast two-hybrid system is an effective method for identifying hepatocyte proteins interacting with F protein of hepatitis C virus. F protein may bind to different proteins.

Quantitative and Qualitative Investigation of Traditional Chinese Medicine Nursing Protocols for Type 2 Diabetes Mellitus

Objective:To analyze the implementation status of traditional Chinese medicine(TCM)nursing protocols for type 2 diabetes mellitus.Methods:Totally 420 hospitalized patients with type 2 diabetes mellitus were given TCM nursing protocols by syndrome differentiation.A quantitative and qualitative investigation was conducted to patients who received TCM nursing and nurses who performed TCM nursing techniques.Results:According to the self-evaluation outcomes of patients,TCM nursing protocols were effective to relieve polydipsia,polyuria,fatigue and other symptoms.Patients exhibited a high level of compliance with nursing techniques such as auricular point pressing,acupoint massage,Chinese herbal foot-bath and moxibustion.The interview showed that patients could benefit from the TCM nursing protocols,and nurses considered that application of TCM nursing protocols could enhance their awareness of TCM knowledge and practice skills.Conclusion:The application of TCM nursing protocols is effective to relieve clinical symptoms of type 2 diabetes mellitus.It can also improve the knowledge level and practice skills of nurses in TCM hospitals.

Reexploration of Structural Changes in Element Bromine through Pressure-Induced Decomposition of Solid HBr

Simple molecular solids have been an important subject in condensed matter physics,particularly for research of pressure-induced molecular dissociation.We re-explore the structural changes of element bromine through pressure-induced decomposition of solid HBr.The phase changes in HBr are investigated by Raman spectroscopy and synchrotron x-ray diffraction up to 125 GPa at room temperature.By applying pressure,HBr decomposes into solid bromine in the pressure range of 18.7-38 GPa.The solid bromine changes from molecular phase to incommensurate phase at 81 GPa,and finally to monatomic phase at 91 GPa.During the process of pressureinduced molecular dissociation, the intermediate incommensurate phase of element bromine is confirmed for the first time from the x-ray diffraction studies.The decomposition of HBr is irreversible since HBr cannot form again upon pressure decompression.

High-Pressure Behavior of Nano-Pt in Hydrogen Environment

We choose nano-Pt in hydrogen environment to explore the size effect on the formation of metal hydrides.At 30 GPa,a phase transition in the metal lattice from the cubic to hexagonal phase is observed characterized by a drastically increased volume per metal atom,indicating the formation of PtH-P63/mmc.We find that nano-Pt could form PtH at a lower pressure than the bulk Pt due to its high specific surface and structure defects.The present work provides the possible route to new metal hydrides under mild conditions.

The Unexpected Stability of Hydrazine Molecules in Hydrous Environment under Pressure

The incomplete decomposition product of metastable hydrazine(N2H4)instead of the energetically favorable ammonia(NH3)upon decompression is one drawback in applications of energetic material oligomeric hydronitrogens.We explore the stability of hydrazine molecules in hydrazine hydrate(N2H4 H2O)under pressure in diamond anvil cells(DACs)combined with in situ Raman spectroscopy and synchrotron x-ray diffraction(XRD)measurements.The results show that one NH2 branch forms NH3 group by hydrogen bonds between hydrazine and water molecules after the sample crystallizes at 3.2 GPa.The strengthening hydrogen bonds cause the torsion of hydrazine molecules and further dominate a phase transition at 7.2 GPa.Surprisingly,the NN single bonds are strengthened with increasing pressure,which keeps the hydrazine molecules stable up to the ultimate pressure of 36 GPa.Furthermore,the main diffraction patterns show continuous shift to higher degrees in the whole pressure range while some weak lines disappear above 8.2 GPa.The present peak-indexing results of the diffraction patterns with Materials Studio show that the phase transition occurs in the same monoclinic crystal system.Upon decompression,all of the hydrazine molecules extract from hydrazine hydrate crystal at 2.3 GPa,which may provide a new way to purify hydrazine from hydrate.

Microsurgical vasoepididymostomy for patients withinfectious obstructive azoospermia, cause, outcome,and associated factors

Azoospermia is of great importance to male infertility. Obstructive azoospermia （OA） due to infection is the most prevalent form of OA in China and has been less studied. We aim to observe the treatment outcome of microsurgical vasoepididymostomy （VE） and also to identify the factors relative to the result after reconstructive surgery. Two hundred and eight men presenting with OA due to infection during the study period from July 2010 to July 2013 were prospectively evaluated. Clinical examination, semen analysis, serum follicle stimulating hormone （FSH）, and scrotal ultrasound were done before surgical exploration. Among the 198 men who were selected for surgical procedures, 159 candidates underwent microsurgical VE with sperm detected in the epididymal fluid. As for the other 39 cases, reconstruction was not feasible. The average age was 28.5 ＋ 3.9 years （range 22-38）, with average follow-up being 16.5 ~ 5.9 months （range 4-28）. According to the 150 cases being followed after VE procedures, the total patency rate was 72% （108/150）. During follow-up, 38.7% （58/150） natural pregnancies occurred, with overall live birth rate being 32.7% （49/150）. Our data suggested that microsurgical VE is an effective therapy for postinfectious epididymal OA. Individualized counseling with prognosis based on etiology should be offered to patients to select optical therapy.

A prospective randomized controlled study on scheduled PDE5i and vacuum erectile devices in the treatment of erectile dysfunction after nerve sparing prostatectomy

Cavernous nerve injury is an important cause of erectile dysfunction(ED).Although protective nerve technology has been widely used in nerve-sparing radical prostatectomy(nsRP),the incidence of ED is still very high after surgery.The purpose of our study was to evaluate erectile function(EF)and penile length in the non-erectile state(PLNES)following scheduled phosphodiesterase 5 inhibitor(PDE5i),vacuum erectile device(VED)treatment,and combination therapy after nsRP.One hundred patients with localized prostate cancer and normal EF were randomized to scheduled PDE5i group,VED treatment group,a combined treatment group,and the control group without any intervention.The International Index of Erectile Function-5(IIEF-5)scores and PLNES were evaluated after 6 months and 12 months of treatment.Sexual Encounter Profile(SEP-Question 2 and SEP-Question 3)were evaluated after 12 months of treatment.Ninety-one of the 100 randomized patients completed the study.We found that the 5 mg tadalafil once a day(OaD)combined with VED can help improve IIEF-5 scores in nsRP patients after both 6 months and 12 months.VED alone or combined with tadalafil OaD can help patients maintain PLNES.VED combined with tadalafil OaD can improve the rate of successful penetration(SEP-Question 2)after 12 months.There were no significant differences in the return to target EF after 12 months among the groups.No significant correlation was noted between the variables and return to target EF(IIEF≥17),and between the variables and effective shortening of the patient’s penis(shortening≥1 cm)after 12 months of intervention.

A rare case of spontaneous renal cholesterol crystallization embolism

To the Editor: Cholesterol crystal embolism (CCE) is a systemic disease caused by cholesterol crystal embolization leading to the occlusion of small arteries in a variety of organs. It may occur spontaneously, or more often, after intravascular procedures such as coronary angiography and carotid artery stenting, cardiovascular surgery such as coronary artery bypass grafting or aortic aneurysm surgery.[1] The kidneys were previously reported to be the most frequent target organ for CCE, and renal CCE was found during the course of clinical illness in approximately 50% of patients with CCE.

Effects of cavitation on vortex-induced vibration of a flexible circular cylinder simulated by fluid-structure interaction method

To investigate the effects of the cavitation on the vortex-induced vibration,the flow and the vibration characteristics of a flexible circular cylinder in the non-cavitating and cavitating flows at Re=200 are simulated using a fluid-structure interaction(FSI)solver,developed in the OpenFOAM framework.The accuracy of the fluid-structure interaction solver is verified using the benchmark cases.For a freely vibrating circular cylinder,the characteristics of the vortex-induced vibration are determined at different frequency ratios.The frequency lock-in is observed in both the cavitating flow and the non-cavitating flow,and the lock-in frequency range is larger in the cavitating flow with a smaller vibration amplitude.In the lock-in frequency range,due to the reduction of the fluid forces and the resonance effects at the dominant frequency,the vibration amplitude is decreased in the cavitating flow,as compared to the non-cavitating flow.Moreover,the low-density vapor and the cavitation collapse weaken the constraints of the flow and the stability of the shedding vortices,respectively,making the flow around the circular cylinder more sensitive to the structural vibration.Consequently,the lock-in frequency range is larger in the cavitating flow.