With the continuous development of modern chemistry, biology and pharmacy, researches on TCM modernization has witnessed a wide range of application of related modern technology, especially in researches of TCM monomer efficacy and pharmacokinetics. This can provide basis to internationalization of traditional Chinese medicine. The current research focus is how to obtain targeted TCM monomer. This article expounded the screening methods for TCM monomer at home and abroad, the principles of screening, the limitation and meanings.

Objective To establish a primary culture of human melanocytes from tiny skin sheets harvested by using a suction blister method, to carry out a serial subcultivation of the melanocytes with human amniotic membrane (AM) as a scaffold, and to observe the influence of AM on the adhesion, proliferation and dendrite development of melanocytes. Methods Tiny skin sheets were collected from the flexual forearm or lower abdomen of a healthy male volunteer by a suction blister method and melanocytes in the skin sheet were counted following Dopa staining under a microscope. The trypsinized skin sheets were scraped with a scalpel to harvest melanocytes which were subjected to a primary culture. Then, the melanocytes were inoculated onto fresh or cryopreserved AM followed by a culture for various durations (4, 8 and 12 days). The morphology and dendrite development of melanocytes were visualized under an inverted microscope after dopa-staining, cell viability evaluated by MTT assay, the adhesion to AM examined by hematoxylin and eosin (HE) staining protocol. Results The density of melanocytes was 1543.1±13.3 cells per mm2 and 857.4±101.7 cells per mm2 in skin sheets obtained from the forearm flexure and lower abdomen of the volunteer, respectively. A skin sheet of about 25.1 mm2 from approximately two blister roof was required to ensure the success of primary culture of melanocytes within 1 month. After culture on fresh or cryopreserved AM for 4, 8, and 12 days, most melanocytes were bi-polar with extended slender dendrites compared with those cultured in common cell culture medium. HE staining showed that melancytes adhered and were evenly distributed on the basement membrane of AM. MTT assay showed that the AM inhibited the proliferation of melanocytes, and no statistical difference was observed in the inhibitory effect between fresh AM and cryopreserved AM (P＞ 0.05). Conclusions Enriched with melanocyes, flexural forearm is a preferable donor site to offer skin sheets for primary culture of melanocytes. Human AM could improve the adhesive growth and dendrite development of melanocytes, and may serve as a promising bioscaffold for in vitro expansion of melanocytes.

Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. Coli and purified by nickel-nitrilotriacetic acid (Ni2+-NTA) affinity resin column,finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.

Objective To study the immunoregulatory activity of eucommia ulmoides oliv polysaccharides (EOP) on immunodepressive mice. Method The immunodepressive mice model was established with cyclophosphamide (CY) treated. EOP was given with different doses, and mouse weight, thymus index, spleen index, abdominal cavity macrophage engulfing rates, engulfing index were measured. Result 300 grams Eucommia ulmoides Oliv crude drugs after withdraw separate gained 9.3 grams polysaccharides. After CY injection, the mouse weight, chest gland index, abdominal cavity macrophage swallowing rate, phagocytic count reduced. EOP could resist the mouse body weight drop induced by CY, elevate the thymus index of immunity low mouse, obviously increase mouse abdominal cavity macrophage engulfing rates and engulfing index (P

Based on the relevant theories and methods concerning the evaluation of higher education, authors used student questionnaires to evaluate the teaching quality of the 23 courses in the postgraduate curriculum, including basic theoretic courses, specialty-related basic courses and courses of methodology for scientific research. The methods used involve 'evaluation by students' and the integrated evaluation of both the quantitative and qualitative aspects of various factors. The easy-to-use student's questionnaires make it possible to find, with respect to students, teachers, and courses, many factors affecting the teaching quality, and at the same time, to provide the close following-up of the process of postgraduate training.

A 73-year-old man developed a skin-colored granule-sized papule on the scrotum 10 years prior to the presentation,which had increased in size and number without obvious symptoms.Histopathological examination showed multiple enlarged intradermal cysts.The cyst wall consisted of one or two layers of flat or columnar cells.Decapitation secretion was seen in the inner layer cells,and the out layer of cysts was formed by flat or cubic myoepithelial cells.A diagnosis of apocrine hidrocystoma of the scrotum is made.

Sequences analysis revealed Grass carp reovirus (GCRV) s10 was 909 nucleotides coding a 34 kDa protein denoted as VP7, which was determined to be a viral outer capsid protein (OCP). To obtain expressed OCP in vitro, a full length VP7 gene was produced by RT-PCR amplification, and the amplified fragment was cloned into T7 promoted prokaryotic expression vector pRSET. The recombinant plasmid,which was named as pR/GCRV-VP7,was then transformed into E.coli BL21 host cells. The data indicated that the expressed recombinant was in frame with the N-terminal fusion peptide. The over-expressed fusion protein was produced by inducing with IPTG, and its molecular weight was about 37kDa, which was consistent with its predicted size. In addition, the fusion protein was produced in the form of the inclusion body with their yield remaining steady at more than 60% of total bacterial protein. Moreover,the expressed protein was able to bind immunologically to anti-his-tag monoclonal antibody (mouse) and anti-GCRV serum (rabbit). This work provides a research basis for further structure and function studies of GCRV during entry into cells.

Objective To assess the relationships among the autologous serum-induced skin wheal-andflare reaction,ex vivo serum-induced basophil histamine release, and serum levels of IgG anti-FcεRI autoantibodies in patients with chronic idiopathic urticaria (CIU).Methods Sixty patients with CIU collected from the Renmin Hospital of Wuhan University were recruited for this study.Sera were obtained from the subjects,and ASST was performed in all of the subjects.The results of ASST were determined according to a recommended criterion described by Sabroe et al,and the positive results were further subclassified into wheal plus flare (W+F) pattern and wheal-only (W) pattern,negative results into flare-only (F) pattern and no response pattern.Enzyme linked immunosorbent assay was used to quantify the content of histamine released by autologous serum-induced basophils and serum levels of IgG anti-FcεRI autoantibodies.Results Of the 60 patients,19 (31.7％)were positive for ASST,including 16 (84.2％) presenting W+F pattern and 3 presenting W pattern; 41 were negative for ASST,including 3 (7.3％) giving F pattern and 38 giving no response pattern.The histamine release rate was significantly higher in ASST-positive patients than in ASST-negative patients (33.38％ ± 9.83％ vs.4.06％ ±1.44％,t =5.13,P＜ 0.01),and was nearly twice as high as that in basophils induced by 10 μmol/L formylmethionyleucylphenylalanine (Fmlp).The serum levels of IgG anti-FcεRI autoantibodies were high in only patients giving (W+F) pattern (757.64 ± 168.99 ng/L),but low in the normal human controls (43.25 ± 16.63 ng/L).Conclusions The positive ASST result of wheal plus flare pattern is associated with high serum levels of IgG anti-FcεRI autoantibodies,and is suggestive of a clinical diagnosis of autoimmune chronic urticaria (ACU).

Objective To investigate the clinical application of statins for secondary prevention and its efficacy in patients with ischemic stroke.Methods A total of 300 patients with ischemic stroke were divided into statins group (n=91) and control group (n=209) according to whether statins was received or not.Serum lipid parameters,liver and kidney function,carotid artery intimamedia thickness (IMT) and plaque area were compared between the two groups.Results In 209 patients not taking statins,68 cases (32.5％) were not advised by clinic doctor; 55 cases (27.8％)were reluctant to take statins in fear of adverse reactions; 77 cases (36.8％) discontinued stains when serum lipids were normal; 3 cases (1.4％) developed skin allergic reaction and the skin rash that disappeared after stains discontinuance; 3 cases (1.4％) had elevated liver enzyme levels and the enzyme levels were declined to normal after stains discontinuance.The levels of total cholesterol (TC),triglyceride (TG) and low density lipoprotein cholesterol (LDL-C) were significantly lower in statin group than in control group [(4.93±0.45) mmol/L vs.(5.87±0.38) mmol/L,(1.68±0.61)mmol/L vs.(2.01±0.58) mmol/L,(2.49±0.57) mmol/L vs.(3.76±0.44) mmol/L,t=18.60,4.46,20.94,P=0.000,0.007,0.000,respectively].High density lipoprotein (HDL-C) level was significantly higher in stain group than in control group [(1.36±0.31) mmol/L vs.(1.03±0.25)mmol/L,t=9.75,P=0.001].There were no significant differences in liver and kidney function parameters such as alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase,serum total bilirubin,direct bilirubin,indirect bilirubin,glutamyl transferase,blood urea nitrogen and serum creatinine between the two groups (all P＞0.05).The carotid IMT and carotid atherosclerotic plaque area were significantly lower in stain group than in control group [(1.62 ± 0.23) mm vs.(1.74±0.31) mm,(0.57±0.12) cm2 vs.(0.65±0.18) cm2,t=3.32,3.88,P=0.008,0.002,respectively].Conclusions Most patients with ischemic stroke are still not taking statin for secondary prevention.Statins can improve the blood lipid levels and carotid arteriosclerosis status,and no obvious adverse reactions are observed.

Objective To investigate the effect of dexmedetomidine on the stress responses during wakeup test in patients undergoing cerebral functional area operation performed under propofol combined with remifentanil anesthesia.Methods Thirty-six ASA physical status Ⅰ or Ⅱ patients,undergoing cerebral functional area operation requiring wake-up test,aged 18-60 yr,weighing 50-70kg,were randomly divided into control group (group C) or dexmedetomidine group (group D) with 18 cases in each group.Dexmedetomidine 0.8 μg/kg was infused over 10 min before induction of anesthesia followed by infusion at 0.4 μg·kg-1 · h-1 in group D,while the equal volume of normal saline was infused in group C.Anesthesia was induced with target-controlled infusion of propofol and remifentanil and iv injection of cisatracurium.At 30 min prior to wake-up test,target-controlled infusion of propofol and application of mulscle relaxants were stopped,the target plasma concentration of remifentanil was decreased to 1 ng/ml,and in group D the infusion rate of dexmedetomidine was decreased to 0.1 μg·kg 1· h-1.Anesthesia time and consumption of anesthetics before wake-up test,wake-up time,and development of complications and intraoperative awareness during wake-up test were recorded.At 30 min prior to wake-up test (T1),immediately after wake-up (T2),at 5 min after wake-up (T3),and at 10 min after the anesthetic depth was deepened (T4),HR,mean arterial pressure and BIS value were recorded and blood samples were taken for determination of plasma concentrations of epinephrine (E) and norepinephrine (NE).Results Compared with group C,the consumption of propofol and remifentanil was significantly reduced before wake-up,the incidence of hypertension was decreased during wake-up test,and HR and plasma E and NE concentrations were decreased at each time point (P ＜ 0.05),and no significant difference in mean arterial pressure and BIS value was found in group D (P ＞ 0.05).Tachycardia,restlessness,bucking and awareness were not observed during wake-up test in group D.Conclusion Dexmedetomidine can inhibit the stress responses during wake-up test and raise the quality of wake-up test in patients undergoing cerebral functional area operation performed under propofol combined with remifentanil anesthesia.