Objective To discuss the importance of the clinic skill learning center in diagnostics teaching.Methods The experiment group was taught in the clinic skill learning center with the reformed method,while the control group was taught in the traditional laboratories with the traditional one.Results The experiment group's two average scores of diagnostics and skill examine are higher than the control group's,and the difference between the two group's average scores are significant in the statistics analysis.Conclusion The applications of the clinical skill learning center in medicine teaching is the need of developing the medical education business and fostering standardized medical talents.

Objective To discuss the meaning of the method of two-step pattern case teaching in image diagnostics. Methods The experiment group was taught by the two-step pattern case teaching,while the control group by the traditional method. Results The experiment group’s image diagnostics exam average score is higher than the control group’s,and the average scores of other major clinical courses are all higher. Conclusion The applications of the two-step pattern case teaching in medicine teaching is the need of developing the medical education business and improving the medical teaching quality.

OBJECTIVE:To provide reference for clinical rational use of anti-epileptic drugs. METHODS:In this retrospective review,serum concentrations of anti-epileptic drugs in a total of 499 patients who were treated with anti-epileptic drugs (such as sodium phenytoin,phenobarbital,carbamazepine and valproate sodium) in our hospital during 2007 were analyzed statistically. RESULTS:Among the patients receiving one kind of anti-epileptic drugs,206 cases (61.49%) were within normal range in blood concentration versus only 45 cases (44.12%) for patients receiving combined drugs. In addition,the above four drugs (sodium phenytoin,phenobarbital,carbamazepine and valproate sodium) were detected in 59.68% of the patients who took Chinese medicines,and among them,3.23% were within normal range in blood concentration. CONCLUSION:Monitoring on serum concentrations of anti-epileptic drugs is conducive to a better control of therapeutic concentration. It is advisable to refrain from using anti-epileptic drugs in combination but to adopt individualized administration. In addition,great importance should be attached to whether there are chemo-synthetic drug components contained in Chinese medicine.

Background The ubiquitin-proteasome pathway(UPP)involves 80%-90% degradation of all in- tracellular proteins.Both ubiquitin and rat component 3 of proteasome(RC3)are hence considered to be central me- diators for cell biology.Objective To evaluate the effect of simvastatin on neointimal hyperplasia and the expres- sion of ubiquitin and rat component 3 of proteasome(RC3)after balloon injury in carotid artery.Methods Thirty- two male SD rats were randomly to receive,low dose(0.4 mg/ng,n=8),or moderate(4 mg/ng,n=8),or high- dose(40 mg/ng,n=8)of simvastatin treatment for 28 days.Common carotid aortic artery was injuried rat ballon. The ratio of intima-media(I/M)thickness was determined.The expression level of ubiquitin and RC3 mRNA were assessed by RT-PCR.The expression level of ubiquitin protein was examined with immunohistochemistry. Results Simvastatin inhibited the expression of ubiquitin and RC3 mRNA and ubiquitin protein in dose dependent manner.The intima-media ratio(-50.2 %)and the expression of ubiquitin(-60.3 %)and RC3 mRNA and ubiq- uitin protein(-60.5 %)was reduced by the high dose simvastatin [40 mg/(kg?d)](P

Objective To investigate the effects of morphine on PTEN expression and NF-κB activity in human gastric carcinoma cell line MGC-803.Methods The human gastric cancer cell line MGC-803 was purchased from Cell Biology Research Institute,Chinese Academy of Sciences,and cultured in DMEM liquid culture medium.The cells were randomly divided into 2 groups(n = 6 each): control group and morphine group.The cells was exposed to 0.1 μmol/L morphine in morphine group.The apoptosis was assessed by flow cytometry after being incubated with morphine for 24 h.PTEN expression and NF-κB activity were detected using RT-PCR and Western blot.Results The apoptotic rate was significantly increased,PTEN expression was up-regulated and NF-κB activity was significantly decreased in morphine group compared with control group(P ＜ 0.05).Conclusion Morphine can promote the apoptosis in human gastric cancer cells by up-regulating PTEN expression and decreasing NF-κB activity.

This study aims to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 using the orthogonal design method, and to investigate its effects of the component formula on cell proliferation, apoptosis and cytoskeleton in lung cancer A549 cells. The orthogonal design method was introduced to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 cells. CCK-8 assay and Real-time cell analysis were adapted to analyze the effect of component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on A549 cells viability at different time and dose. Cell apoptosis was measured by Annexin V- FITC/PI double staining and flow cytometry. Cell skeleton protein F-actin was detected by high content screening (HCS). The optimizing component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma for total salvianolic acid, total saponins of panax ginseng and ginseng polysaccharide doses were 5, 10, 5 mg L(-1). CCK-8 assay and real-time cell analysis demonstrated that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma treatment could significantly decrease the A549 cell viability in both dose- and time-dependent manner compared with control group (P < 0.01). Moreover, the increase of cell apoptosis was detected by Annexin V-FITC/PI double staining and flow cytometry when cells treated with the component formula, which indicating that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma could induce A549 cell apoptosis in a time-dependent manner compared with control group (P < 0.01). Furthermore, compared with control group, a significant decrease in A549 cell skeleton area was found in the component formula-exposed cells in the dose-dependent manner (P < 0.01). In summary, the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma inhibits A549 cell proliferation by inducing cell apoptosis and decreasing cell microfilament formation. All of these results will be helpful to reveal antitumor mechanism of the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma, which provides a basis for the exploration of antitumor mechanism of the component formula on lung cancer.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; Panax ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; Plant Roots ; chemistry ; Rhizome ; chemistry ; Salvia miltiorrhiza ; chemistry

Natural products are important sources of drug discovery. However, the traditional methods of extraction and isolation, as well as chemical synthesis for obtaining natural products are associated with issues such as operational complexity, high costs, low efficiency, and environmental pollution. Constructing microbial cell factories through synthetic biology methods to produce medicinal natural products has the advantages of high efficiency, low cost, and environmental protection. Nevertheless, the scope and yield improvement of the products are limited by the limitations of enzymes in microbial cell factories. Protein engineering is considered one of the most effective approaches to overcome these limitations. This article introduces commonly used methods of protein engineering technology and summarizes its specific applications in improving enzyme performance, modifying the enzymatic environment, and promoting the development of synthetic biology tools in the field of pharmaceutical natural product synthesis. Furthermore, it analyzes the current bottlenecks and challenges in protein engineering and looks forward to its future application prospects, offering insights for the development and practical use of protein engineering technology.

Objective To discuss the ultrasound imaging of acardia twin before and after spontaneously blood blocked in twin reversed arterial perfusion sequence (TRAP), and to analyze the inlfuence factor for the prognosis of pump twin. Methods Seven TRAP with pump twins and acardia′s blood lfow blocked were diagnosed by US and autopsy in Hubei Women and Children′s Hospital between 2001 January to 2012 September. The ultrasound images, clinical data and pump twin’ outcome were analyzed. Results Ultrasound images showed skin edema, acardia, spine and lower limbs in seven acardia cases before blood blocked, among which 4 were acardius acephalus, 3 were acardius anceps. Single umbilical artery were detected in 7 cases with reversed umbilical artery perfusion toward fetus. Four cases had rare blood lfow (UA-PI ratio<0.7. Five acardia cases had slower growing rate than the pump twin. Ultrasound images showed no growing, no blood lfow in the acardia, the acardia twin became vagued in the second and third trimester, and ifnally developed into a paper fetus. A linear umbilical cord extend from umbilical region to placenta were found in 7 cases whose blood lfow were spontaneously blocked. Pump twin′s outcome:3 pump twins survived, 3 died in uterus, and 1 was induced labor due to hydrocephalus and cardiac failure. The most important factors that affected the prognosis of pump twin included:faster growing in acardia twin with less blood supply, abnormal brain and heart function in pump twin, chromosome abnormalities, abnormal amniotic fluid and cord entanglement. Conclusions Less blood flow and slow growth speed prompt in acardia twin suggested spontaneously acardia blood block. It is mandatory to monitor the pump twin after the spontaneous block of acardiac’s blood lfow.

Background Corneal neovascularization (CNV) is one of the causes of corneal blindness.Studies showed that ADP-ribosylation factor (ARF) can regulate the growth of tumor cells,and inhibiting ARF will decrease angiogenesis.However,whether ARF antagonist plays an action on CNV is unclear.Objective The aim of this study was to explore the effect and mechanism of ARF inhibitor on alkali-burn induced CNV.Methods Sixty clean male BABL/c mice aged 7-8 weeks were divided into PBS group and ARF antagonist group according to randomized number table.CNV models were induced by NaOH burn method in all the mice.ARF at the concentration of 0.5 g/L(0.5 ml) was intraperitoneally injected 3 times per week for 1 week followed the induction of CNV in the ARF antagonist group,and 0.5 ml PBS was used in the PBS group.CNV was examined 2,4,7,14 days after injection by the slit lamp microscope and the CNV related area in the cornea was calculated.Betore modeling(0 day) and 4,7,14 days after modeling,real-time PCR and Western blot were used to analyze the expressions of ARF mRNA and protein in the corneas.Forteen days after modeling,the expression of the CD31 in the CNV was detected using immnofluorescence of corneal whole mount;the expression of vascular endothelial growth factor(VEGF) in the cornea was assayed by Western blot.Cellular wound scratch test was employed to evaluate the effects of ARF antagonist on proliferation and migration of human retinal vascular endothelial cells (RECs).All animal experiments were done in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and Guideline for the Care and Use of Laboratory Animals on the the Soochow University Animal Care Committee.Results ARF mRNA and protein were expressed in the mice corneas in both the PBS group and the ARF antagonist group at various time points.The expression of ARF mRNA in mice corneas was enhanced with the lapse of the time (Ftime =65.17,P =0.00),but no significant difference was found among the groups (Fsroup =1.98,P=0.18).There was also significant difference in the expression of ARF protein in mice corneas at different time points in the ARF antagonist group (F =10.77,P =0.00).The related CNV area was 0.45±0.05 in the ARF antagonist group,and that in the PBS group was 0.72±0.11,with significant difference between them (t =-3.87,P ＜ 0.05).The green fluorescence area of C D31 expression in the cornea was smaller in the ARF antagonist group than that of the PBS group.Expression level of VEGF in the ARF antagonist group was 1.20±0.21,and that in the PBS group was 2.47±0.33,showing a significant difference (t =-5.62,P ＜ 0.05).As the increase of ARF antagonist concentration,the inhibiting rate of cell proliferation was reinforced among 10,100 and 1 000 μg/L ARF antagonist groups (F=8.47,P =0.02).Twenty-four hours after scratch test,the migrating distance of human RECs was (5.46±1.32) μm and (5.04±1.68) μm in the 100 μg/L and 1 000 μg/L ARF antagonist groups,respectively,which were shorter than (8.49± 1.18) μm of the PBS group (t=-2.94,-2.91,both at P＜0.05).Conclusions ARF inhibitor can reduce CNV by down-regulating the expression of VEGF in alkaline burn cornea and inhibiting the proliferation and migration of vascular endothelial cells.

Objective To evaluate the effect of gamma irradiation on the proliferation,differentiation,and mineralization of murine osteoblastic cells,and to investigate the related molecular mechanism.Methods Osteoblastic cells were irradiated by different doses (0,0.5,1.0,2.0,5.0 Gy)of 137Cs γ-rays.Cell morphology was observed with a microscopy,cell viability was analyzed by MTT assay,and ALP activity was analyzed by the methods of enzyme histochemistry and PNPP.Meanwhile,gene expressions of ALP,osteocalcin (OC),collagen Ⅰ,osteoprotegerin (OPG) and receptor activator of nuclear factor-kB ligand (RANKL) were measured by semi-quantified RT-PCR.Results Cell viability decreased with the radiation doses over 1.0 Gy ( t =6.197 - 18.677,P ＜ 0.05 ).After radiation with a dose over 2.0 Gy,the cell number and the junctions of cell protrusions decreased,the cells had low refractivity and the activity and mineralization ability of ALP were also inhibited ( t =2.790 -2l.374,P ＜0.05).In addition,the expressions of ALP and OC mRNA were down-regulated significantly (t =3.563 -16.508,P ＜ 0.05) when the radiation dose was higher than 0.5 Gy,and the expressions of OPG,OPG/RANKL mRNA were down-regulated ( t =12.942,4.954,P ＜ 0.05 ) at 5 Gy.But the expressions of collagen Ⅰ and RANKL mRNA were not affected by irradiation.Conclusions The osteoblastic cells were significantly influenced by γ-irradiation,including morphological changes,inhibition of cell proliferation,differentiation and mineralization ability. Meanwhile,mRNA expressions of ALP and OC were downregulated.OPG/RANKL may be a main pathway of osteoblastic cell damage under high dose radiation.