1.High-throughput Drug Screening Method and Application for TCM Monomer
Chinese Journal of Information on Traditional Chinese Medicine 2014;(8):4-6
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.
2.Culture of human epidermal melanocytes with human amniotic membrane as a scaffold and their biological ability identification
Xiaoguang SHE ; Xiaoming LIU ; Tiechi LEI
Chinese Journal of Dermatology 2011;44(2):117-120
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.
3.Herpes Simplex Virus Type 1 ICP27 Protein: Its Expression, Purification and Specific Antiserum Production
Lei ZHAO ; Xiaoming REN ; Alan C.ZHENG
Virologica Sinica 2010;25(3):199-205
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.
4.Research on the Evaluation Model Concerning Teaching Quality of Courses for Medical Postgraduates
Lei TANG ; Xiaoming CHEN ; Guoyao TANG ;
Chinese Journal of Medical Education Research 2002;0(01):-
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.
5.Effect of Immunoregulatory Activity of Eucommia Ulmoides Oliv Polysaccharides on Immunodepressive Mice
Xiaoming XIN ; Hao WANG ; Lei FENG
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(10):-
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
6.A case of apocrine hidrocystoma of the scrotum
Xiaoming QIN ; Lei XU ; Li WU ; Lei KONG
Chinese Journal of Dermatology 2012;45(9):672-673
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.
7.The effect of tourniquet compression on axonal transport in sciatic nerve of rats
Lei WANG ; Ran XIAO ; Ye WANG ; Xiaowen LIU ; Xiaoming DENG
Chinese Journal of Anesthesiology 2011;31(3):289-291
Objective To investigate the effect of tourniquet compression on axonal transport in sciatic nerve of rats.Methods Twenty-four 12-week old male SD rats weighing 250-300 g were randomly divided into 4groups according to the duration of tourniquet compression(n=6 each):1,2,4 and 12 h.The tourniquet was applied to the middle 1/3 of thigh.In each animal whether the left or right thigh was compressed was determined by a flip of coin.The tourniquet was released for 10 min after every hour of compression.A 3-cm segment of sciatic nerve was removed at the end of tourniquet compression(1.5 cm proximal and 1.5 cm distal to the site of compression).Immuno-histochemistry was used to measure the expression of insulin-like growth factor-1(IGF-1)in the sciatic nerve.The ratio of average optic density of the compressed sciatic nerve to that of control was used to estimate the degree of IGF-1 accumulation.The regression equation of the interaction between the duration of compression and accumulation of IGF-1 was analyzed.Results There was significant accumulation of IGF-1 in the sciatic nerve proximal to the compressed site.The accumulation increased with the duration of compression.There was no significant accumulation of IGF-1 in the sciatic nerve distal to the compressed site.The regression equation of the interaction between the duration of compression(X)and accumulation of IGF-1(Y)was Y=0.422X+0.887.Conclusion Tourniquet compression of sciatic nerve can inhibit axonal transport.The accumulation increases with the duration of compression.
8.Clinical significance of autologous serum skin test (ASST) in the diagnosis of chronic urticaria
Xiaolei HE ; Tiechi LEI ; Xiaoming LIU ; Zhifeng FAN ; Ying SHI
Chinese Journal of Dermatology 2012;45(1):5-8
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).
9.Effect of dexmedetomidine on stress responses during wake-up test in patients undergoing cerebral functional area operation performed under propofol combined with remifentanil anesthesia
Zhenni ZHANG ; Jianrui LU ; Yong ZHANG ; Xiaoming LEI ; Rongliang XUE
Chinese Journal of Anesthesiology 2013;33(8):951-954
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.
10.Effects of high dose atorvastatin administration on platelet activities and ventricular remodeling of patients with acute myocardial infarction
Shaonan LI ; Zhen LIU ; Huifang HUANG ; Pingan CHEN ; Xiaoming LEI
Journal of Chinese Physician 2015;17(4):519-523
Objective To investigate the effects of high dose atovastatin administration on platelet activity and ventricular remodeling of patients with ST-Segment elevation myocardial infarction (STEMI) underwent primary percutaneous coronary intervention (PCI).Methods A total of 260 STEMI patients who hospitalized in our Department of Cardiology from June 2012 to December 2013 was enrolled and randomly divided into two groups:controlled group (n =140) and high dose atorvastatin group (n =120).Indicators of platelet activities including mean platelet volume (MPV),platelet large cell ratio (P-LCR),blood CD62p,and glucose protein Ⅱ b/Ⅲa (PAC-1) were measured before and 48 hours after PCI.TIMI myocardial perfusion grade (TMPG) after PCI was recorded and patients accepted ultrasound cardiogram (UCG) examinations 5 ~7 days after PCI and 6 months after discharge.After PCI,Patients were followed up for 6 months,statin-associated liver impairment,myopath and major adverse cardiac events (MACE) happened during follow-up periods were recorded.Results MPV,P-LCR,CD62p,and PAC-1 in patients of high dose atorvastatin group were less than controlled group and TMPG were better than controlled group [(12.96±1.73)fl vs (14.18 ± 1.86)fl,P <0.05;(29.12 ±5.83)% vs (30.66 ±6.12)%,P < 0.05;(45.36±5.24)% vs (48.44±4.75)%,P <0.01;(74.61 ±5.57)% vs (78.55±5.78)%,P <0.01].Six months after PCI,UCG examination showed that Left ventricular end-diastolic volume (LV-EDV),left ventricular end-systolic volume (LVESV) and left ventricular mass index (LVMI) in high dose group were less than controlled group while the left ventricular ejection fraction (LVEF) was higher than controlled group [(110.46 ±8.86)ml vs (112.61 ±8.5)ml,P <0.01;(60.16 ±6.13)ml vs (63.52 ± 5.54)ml,P <0.01;(1O1.69±4.35)g/m2 vs (103.96 ±4.17)g/m2,P <0.05;(50.08 ±3.78)% vs (48.47 ± 4.12) %,P < 0.05].After 6 months of follow-up,the incidence rate of statin-associated liver impairment and myopathe had no significant difference between two groups and Kaplan-Meier survival analysis showed patients of two groups had significantly different cumulative non-events survival rates (91.7% vs 82.4%,Log rank =4.409,P =O.036).Conclusions Loading dose atorvastatin before PCI combined high maintenance dose after PCI can inhibit platelet activation and improve myocardial perfusion levels of patients with STEMI underwent primary PCI.It also can reduce Left ventricular remodeling and improve patient's prognosis without increasing side effects.