重组人睫状神经因子的表达、纯化以及聚乙二醇修饰

目的 研究重组人睫状神经因子(rh-CNTF)的表达、纯化及聚乙二醇化修饰、纯化方法.方法 重组人睫状神经因子(rh-CNTF)工程菌经37℃培养至对数生长期,加入IPTG诱导4 h,经Q Sepharose F.F凝胶柱纯化和复性后得到纯化的rh-CNTF.以体外培养鸡胚背根神经节(DRG)方法测定重组蛋白rh-CNTF的神经营养活性.用单甲氧基聚乙二醇丁醛(mPEG-ButyrALD-20 k)对rh-CNTF进行修饰,所得修饰产物经DEAE Sepharose F.F和Superdex 75凝胶分离纯化后获得mPEG-rh-CNTF偶联物.结果 目的 蛋白占总蛋白30%左右,为包涵体,经Q Sepharose F.F凝胶柱纯化和复性,获得纯度达90%的rh-CNTF.表达的rh-CNTF能促进培养的鸡胚背根神经节神经突起的生长.修饰后的单聚PEG-rhCNTF经DEAE Sepharose F.F和Superdex 75凝胶纯化后纯度达到95%.结论 确立了rh-CNTF蛋白表达纯化及PEG修饰的方法.     

C型产气荚膜梭菌重组菌株BL21(DE3) pXETAB2B1表达条件的优化

目的 优化C型产气荚膜梭菌重组菌株BL21(DE3)pXETAB2B1融合蛋白表达的条件.方法 以IPTG(异丙基硫代-β-D-半乳糖苷)为诱导剂,通过聚丙烯酰胺凝胶电泳比较pH值、培养温度、诱导时间、菌体生长密度等因素对重组α-β2-β1融合蛋白表达进行表达条件的影响.结果 C型产气荚膜梭菌重组菌株BL21(DE3)pXETAB2B1的表达优化条件是最适培养基pH值是7.5,最适诱导时间是5小时,最适表达温度是37℃,最适菌体生长密度是OD6001.0,IPTG最适诱导浓度是0.4 mmol/L.结论 获得C型产气荚膜梭菌重组菌株BL21(DE3)pXETAB2B1融合蛋白的最佳表达条件.     

C型产气荚膜梭菌重组菌株B121（DE3）pXETAB2B1表达条件的优化

目的优化C型产气荚膜梭菌重组菌株B1221（DE3）pXETAB2B1融合蛋白表达的条件。方法以IPTG（异丙基硫代-β—D-半乳糖苷）为诱导剂，通过聚丙烯酰胺凝胶电泳比较pH值、培养温度、诱导时间、菌体生长密度等因素对重组α-β2-β1融合蛋白表达进行表达条件的影响。结果C型产气荚膜梭菌重组菌株BL21（DE3）pXETAB2B1的表达优化条件是：最适培养基pH值是7．5，最适诱导时间是5小时，最适表达温度是37℃，最适菌体生长密度是OD6001．0，IPTG最适诱导浓度是0．4mmol／L。结论获得C型产气荚膜梭菌重组菌株BL21（DE3）pXETAB2B1融合蛋白的最佳表达条件。     

茶树菇中降压活性蛋白提取工艺优化

目的 茶树菇蛋白具有体外降压活性,以水浸提法提取茶树菇降压活性蛋白、优化提取工艺.方法 采用单因素实验考察了液固比、提取时间、提取温度和提取液的pH等因素对茶树菇降压活性蛋白提取率及提取产物对血管紧张素转化酶(ACE)体外抑制效果的影响.结果 降压活性蛋白提取率及浸提产物对ACE抑制效果最佳条件是:液固比为40:1,提取时间3 h,提取温度为30℃.提取液初始pH为8.0,此时活性蛋白质得率最高可达到87.7%.提取物对ACE的抑制活性可达54.O%.结论 本提取工艺是一种操作简单、效率较高的茶树菇降压活性蛋白提取工艺.     

单胺氧化酶抑制药物体外筛选模型的建立

目的：构建一种能用于治疗神经退行性变疾病单胺氧化酶（monoamine oxidase,MAO）抑制药物的体外筛选模型。方法：采用紫外分光光度方法,以苄胺或5-羟色胺（5-hydroxytryptamine,5-HT）分别作为MAO-B或MAO-A的反应底物,测定其活性,并对MAO浓度、底物浓度、缓冲液浓度、pH值、反应时间和温度等进行优化。结果：研究确定的模型反应体系条件为：在温度37 ℃、酶预孵育时间20 min和反应时间60 min下,测定MAO-B活性最佳条件为100 mmol/L磷酸钾缓冲液（pH 7.6）,MAO和底物苄胺的终浓度分别为0.15 mg/ml和2.00 mmol/L;测定MAO-A活性最佳条件为100 mmol/L磷酸钾缓冲液（pH 7.4）,MAO和底物5-HT的终浓度分别为0.40 mg/ml和5.00 mmol/L。结论：建立了一种MAO抑制药物体外筛选的模型,该模型具有成本低、操作简便等特点,适合于大规模筛选MAO-B和MAO-A的抑制药物。     

重组人酸性成纤维细胞生长因子脂质体的制备及其物理稳定性评价

［摘 要］目的：确立重组人酸性成纤维细胞生长因子（rhaFGF）脂质体制备的最佳处方及工艺条件,研究其理化性质。方法：采用冻干再水化法,以磷脂与胆固醇质量之比、缓冲溶液pH值和均质时间为单因素,每个因素选取三水平进行实验,以药物包封率、物理稳定性和生物活性作为评价指标,确立优化脂质体制备的最佳处方及工艺条件。结果：采用最佳处方工艺条件,即磷脂与胆固醇的比例为20∶1,磷酸盐缓冲液pH值为6.5,高压均质时间为20 min制备的rhaFGF脂质体,经测定其包封率（En）达81.79%±2.51%,物理稳定性参数（KE）为1.050%±0.342%,生物学活性为（6.521 0±0.617 1)×10⁵ Ｕ/mL,粒子大小较均匀,粒径大多分布在100 nm左右。结论：通过优化工艺条件制备的rhaFGF脂质体包封率高,在4℃保存条件下,渗漏率低且具有很好的物理稳定性及生物学活性。     

人可溶性TRAIL蛋白表达及纯化条件优化

目的：优化人可溶性TRAIL蛋白生产菌发酵及纯化条件,从而提高TRAIL的产量及活性。方法①探索6种不同培养液、培养液pH值、异丙基硫代半乳糖苷（ IPTG）诱导浓度及时间,选择最佳发酵条件;②联合镍离子亲和层析及超滤法纯化人可溶性TRAIL蛋白。结果①人可溶性TRAIL的最佳发酵条件为pH7．4的YPD培养液,所用IPTG的最佳浓度为0．2 mmol/L,最佳诱导时间为6 h;②经镍离子亲和层析和超滤联合作用可得纯度极高的蛋白。结论成功完成了人可溶性TRAIL发酵及纯化条件的优化,为今后大量提取TRAIL蛋白奠定了基础。     

灵芝锰过氧化物酶毕赤酵母工程菌培养条件的优化

目的 研究毕赤酵母生产重组灵芝锰过氧化物酶(recombinant Ganoderma lucidum manganese peroxidase,rG1MnP)的最优培养条件.方法 在单因素实验基础上,采用中心组合设计和响应曲面法对rGlMnP工程酵母产rGlMnP的发酵条件进行优化,探讨初始pH、温度、甲醇浓度和血红素浓度对重组毕赤酵母生产的rGlMnP的酶活力的影响,分析上述4个因子的交互作用及其最佳水平范围,并验证模型的有效性.结果 初始pH、温度、甲醇浓度和血红素浓度对毕赤酵母生产rGlMnP的酶活力有显著影响;毕赤酵母生产rGlMnP最优的培养条件为:初始pH5.5、诱导温度30℃、甲醇浓度1.5％以及血红素浓度1.0 mmol/L.结论 本研究为利用毕赤酵母工程菌大规模生产rGlMnP奠定了基础.     

探针药物法评价柚皮苷对人肝微粒体CYP2E1酶代谢活性的影响

目的：考察柚皮苷对CYP2E1酶代谢活性的影响,以探讨与其它以CYP2E1为主要代谢酶的药物联用时,是否可能产生潜在的相互作用.方法：在人肝微粒体代谢体系中,以氯唑沙宗作为底物探针,建立表征CYP2E1酶代谢活性的HPLC检测方法,分别考察体外代谢体系的最适宜底物浓度、代谢时间、pH值以及孵育温度.在确定的条件下,将柚皮苷稀释成不同浓度,分别与氯唑沙宗共同孵育于人肝微粒体代谢体系中,测定在有或无柚皮苷存在下氯唑沙宗的剩余量,以评估柚皮苷对CYP2E1酶代谢的影响.结果：在人肝微粒体孵育体系中,氯唑沙宗代谢最适宜的体外代谢条件为底物浓度2.4 g/L,代谢时间3.5h,pH7.5,孵育温度37℃.柚皮苷对CYP2E1酶代谢活性有一定的抑制作用,IC50为10.07 μmol/L.结论：柚皮苷对经CYP2E1酶代谢的药物可能会产生微弱的药物相互作用.     

酶法提取女贞子多糖及其抗氧化性研究

目的 酶法优化女贞子多糖提取工艺.方法 利用纤维素酶处理女贞子提取多糖,分别以酶解时间、酶解温度、酶量和pH值为主要影响因素,采用L9(34)正交实验设计进行提取工艺的优化.结果 通过正交试验及验证性实验确定其最佳提取工艺条件为酶解时间120 min,酶解温度40℃,酶量1％,pH值为5.2.在此条件下,女贞子多糖提取率可达6.58％.结论 酶法提取的女贞子多糖具有显著的抗氧化活性,对DPPH自由基、羟自由基和超氧阴离子自由基作用明显,是一种良好的天然抗氧化剂.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

Dr.Zhang Ren''''s Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

Luo Lingjie''''s Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.