培养基及培养条件对普鲁兰酶的影响

从土壤中分离出一株产普鲁兰酶的菌株,初步鉴定为芽孢杆菌．通过优化发酵培养基及发酵培养条件,在２５０ｍＬ的摇瓶中可达到４．５μｍｏｌ／（ｍｉｎ·ｍＬ）的普鲁兰酶酶活．优化后的发酵培养基成分如下：玉米支链淀粉２ｇ／ｄＬ,蛋白胨２ｇ／ｄＬ,牛肉膏１．５ｇ／ｄＬ,ＮａＣｌ０．５ｇ／ｄＬ,ＭｎＳＯ４２μｍｏｌ／Ｌ．摇瓶发酵工艺条件：接种量１７％,温度３７℃,摇瓶转速２２０ｒ／ｍｉｎ,ｐＨ６．０,发酵周期３２ｈ．通过２Ｌ容积的发酵罐批式发酵,酶活可稳定在３．３μｍｏｌ／（ｍｉｎ·ｍＬ）左右     

发酵生产谷氨酰胺转胺酶的摇瓶条件

在摇瓶条件下， 以淀粉为碳源，蛋白胨及酵母膏为氮源，对Ｓｔｒｅｐｔｏｖｅｒｔｉｃｉｌｌｉｕｍｍ ｏｂａｒａｅｎｓｅ生产谷氨酰胺转胺酶（ＭＴＧ）的摇瓶发酵条件进行了探索．研究结果表明，发酵法生产谷氨酰转胺酶的适宜初始ｐＨ 值、淀粉质量浓度、接种量分别为６．１，２ ｇ／ｄＬ，１０％ ；ＭＴＧ酶活最高可达１．８３ ｕｍ ｏｌ／（ｍ ｉｎ·ｍ Ｌ）．初始加入硫酸铵对菌体生长影响不大，但对ＭＴＧ 的合成却产生抑制作用；初始加入纯氨基酸及胱氨酸母液不利于ＭＴＧ 酶活的提高．     

利用稻谷壳水解液培养单细胞蛋白

以树状假丝酵母１．２５７为菌种，稻谷壳酸水解液为碳源，氨水中和水解液产物为氮源，在摇瓶条件下和１０Ｌ气升式发酵罐上进行发酵工艺条件的研究。结果表明，该菌种在初糖质量浓度２．５ｇ／ｄＬ，碳氮比３∶１，ｐＨ４．５～５．０的培养基中培养４８ｈ可获得较高的菌体蛋白量，在１０Ｌ发酵罐中试验，菌体干重为１２．６ｍｇ／ｍＬ，蛋白质含量为６２．８％，总糖利用率为８５．７％。     

真养产碱杆菌利用葡萄糖生产聚β-羟基丁酸的摇瓶发酵条件

以葡萄糖为碳源，对真养产碱杆菌生产ＰＨＢ的摇瓶发酵条件进行了探索。研究结果表明，在摇瓶补料条件下，细胞干重和ＰＨＢ质量浓度可分别达到３５．１ｇ／Ｌ和２８．３ｇ／Ｌ，ＰＨＢ对葡萄糖的产率系数为０．３６ｇ／ｇ．     

发酵法制核黄素

以实验用Ｅ．ａｓｈｂｙｉＡｓ２．４８１为对象，研究了菌种特性，并对发酵培养基配方及摇瓶工艺条件进行了优化。研究结果：发酵培养基以葡萄糖、蛋白胨、玉米浆为主要原料；发酵起始ＰＨ＝６．８；５００ｍｌ和２５０ｍｌ三角瓶最佳装液量分别是６０～７０ｍｌ和２０～３０ｍｌ；接种量为１０％，在振动次数１００ｒ／ｍｉｎ，振幅８～１０ｃｍ的摇床上进行摇瓶。以豆油或油酸钠为促生因子，核黄素产量从０．１ｇ／Ｌ提高到０．６ｇ／Ｌ．     

L-异亮氨酸的发酵条件

从黄色短杆菌（Ｂｒｅｖｉｂａｃｔｅｒｉｕｍｆｌａｖｕｍ）ＡＴＣＣ１４０６７出发，选育获得一株Ｌ－异亮氨酸高产菌ＺＱ－４．试验结果证明，生物素是ＺＱ－４菌株唯一必需生长因子，生物素或玉米浆用量分别为５０μｇ／Ｌ和２０ｍＬ／Ｌ时可得到较高的产酸率。为了获得较高的Ｌ－异亮氨酸积累量，培养基中葡萄糖质量浓度为１４０ｇ／Ｌ，硫酸铵用量为４０ｇ／Ｌ时较为适宜；以适当的摇瓶装液量改善通风状况可明显提高产酸率。在适宜的发酵条件下，ＺＱ－４经摇瓶发酵培养３ｄ，Ｌ－异亮氨酸积累量可达２８～３０ｇ／Ｌ．     

里氏木霉 Rut C-30 液态发酵法生产纤维素酶

探讨了增强里氏木霉ＲｕｔＣ－３０产纤维素酶的方法，添加葡糖糖于培养基中，可促进菌体生长，但不能提高产酶；采用Ａｖｉｃｅｌ与麸皮复合碳源，以及使用ＫＨ２ＰＯ４－Ｋ２ＨＰＯ４缓冲系统控制发酵液ｐＨ，在摇瓶发酵条件下，可获得很高活力的纤维素酶，培养６ｄ，酶活可达到ＣＭＣａｓｅ１６６７～２０８４ｎｍｏｌ·ｓ－１·ｍｌ－１，ＦＰＡ１５０～２００ｎｍｏｌ·ｓ－１·ｍｌ－１．采用２．５Ｌ发酵罐培养，通过控制ｐＨ和溶氧，纤维素酶活力为ＣＭＣａｓｅ２２２３．８ｎｍｏｌ·ｓ－１·ｍｌ－１，ＦＰＡ１９４．５ｎｍｏｌ·ｓ－１·ｍｌ－１．     

绝经后女性类风湿性关节炎患者腰椎和股骨近端骨密度测量

为了观察绝经后女性类风湿性关节炎（ＲＡ）患者骨矿物质密度水平的变化，采用双能Ｘ线骨密度仪（ＤＥＸＡ）测量２３例绝经后女性ＲＡ患者和年龄、性别匹配的２３例正常健康者的腰椎２～４（Ｌ２～Ｌ４）和右侧股骨近端（股骨颈、大转子和Ｗａｒｄ＇ｓ三角区）的骨密度（ＢＭＤ）。结果表明，绝经后女性ＲＡ患者Ｌ２～Ｌ４、右侧股骨近端的ＢＭＤ测量值（ｇ／ｃｍ２）明显低于年龄和性别匹配的正常健康者（Ｐ＜０．０５）。绝经后女性ＲＡ患者Ｌ２～Ｌ４骨密度的Ｚ－ｓｃｏｒｅ与糖皮质激素用药时间呈显著负相关（ｒ＝－０．５８，Ｐ＝０．００３，ｎ＝２３）。提示绝经后女性ＲＡ患者存在明显的骨质丢失。     

β-葡萄糖苷酶产生菌的选育

对β－葡萄糖苷酶活力为０．４９７μｍｏｌ／（ｓ·ｇ）的无花果曲霉１０１菌株进行Ｃｏ６０及ＵＶ和ＮＴＧ诱变，得到一株产酶提高１．７２倍的Ｌ２菌株；再经发酵培养基及培养条件，包括碳源、氮源、有机酸、表面活性剂、金属离子、产酶诱导剂、培养温度、水分、初始ｐＨ值、通风量、接种形式及接种量等的优化，以优化得到的最佳条件固态发酵６ｄ，产酶稳定在４．１７～４．６７μｍｏｌ／（ｓ·ｇ）．     

多囊卵巢综合征患者外周血生长激素水平测定

为了解多囊卵巢综合征肌（ＰＣＯＳ）患者生长激素（ＧＨ）水平，对２１例（肥胖１０例，非肥胖１１例）ＰＣＯＳ患者进行外周血ＧＨ测定，并以２６名（肥胖１２名、非肥胖１４名）正常月经周期妇女作对照。结果：（１）ＰＣＯＳ患者ＧＨ含量（肥胖组０．９１±０．３５ｐｇ／Ｌ，非肥胖组１．３４±０．４９μｇ／Ｌ）较相应正常对照组（肥胖组１．８６±０．５８μｇ／Ｌ，非肥胖组２．１１±０．８８μｇ／Ｌ）降低（Ｐ＜０．０１，Ｐ＜０．０５）。（２）ＰＣＯＳ患者肥胖组较非肥胖组ＧＨ明显降低。认为ＰＣＯＳ患者伴ＧＨ相对不足，可能与下丘脑生长抑素活性、肥胖和高胰岛素血症有关     

What should be the antibiotic prescription protocol for burn patients admitted in the department of burns,plastic and reconstructive surgery

This is a prospective study with the aim to determine specific patterns of burn wound bacterial colonisation and antimicrobial resistance profiles. There is a high incidence of infections and septicaemia in post‐burn patients, which, in turn, are associated with high morbidity and mortality, a fact that compelled us to undertake this study. The study was conducted over a period 11 months, from 1 August 2014 to 30 June 2015, in 50 burn patients admitted in our burn unit. Wound cultures were taken after 72 hours of admission from all the patients, and then, empirical systemic antibiotics were administered. For wound cultures; 1 cubic cm tissue was taken and placed in aerobic and anaerobic culture vials and transported to the microbiology lab under all aseptic precautions as soon as possible. At the time of fever any time after 72 hours of admission, 16 ml of blood was drawn under all aseptic precautions. Both aerobic and anaerobic blood culture vials were filled with 8 ml of blood each and transported to the microbiology lab. The results of culture and sensitivity reports of 50 patients were recorded. The data obtained was analysed using appropriate statistical analytical tests. The most common organism responsible for bacteraemia is Pseudomonas (43%). Most of the strains of organisms isolated were resistant to commonly used antibiotics in the hospital; Pseudomonas was found 100% resistant to a combination of ampicillin + sulbactum, ceftriaxone and was most often sensitive to imipenem, amikacin and vancomycin. Methicillin‐resistant Staphylococcus aureus (MRSA) was also found resistant to commonly used antibiotics like ceftriaxone, ampicillin + sulbactum and ceftazidime + calvulanic acid. Linzolid and vancomycin were effective in 83% and 100% cases, respectively. We conclude that similar institution‐specific studies should be conducted, and such studies will be helpful in providing useful guidelines for choosing effective empirical therapy that will have a great impact on the prevention of infection and its complications in burn patients because of bacteraemia.     

退变腰椎间盘髓核细胞立体培养与单层培养的生物活性比较

目的：比较体外单层培养和旋转微载体立体培养人退变腰椎间盘髓核细胞的生物学活性指标,探讨更加有效的椎间盘髓核细胞体外培养方法.方法：对获取的腰椎间盘突出症患者的24个椎间盘按年龄分为A组（20～25 岁）、B组（26～30 岁）、C组（36～45 岁）及D组（＞45岁）,分别利用酶消化法进行单层细胞培养和旋转微载体立体培养系统进行立体培养,观察细胞生长形态,检测细胞生长曲线及速度、细胞生长活性、细胞分裂指数及胶原含量.结果：单层培养的髓核细胞贴壁后为多角形或梭形,伸出伪足;立体培养的细胞在微载体上呈梭形或不规则形,呈立体状生长.立体培养的细胞生长速度较快,1周内两种培养方法各时间点及各组间比较均具有统计学意义（P＜0.05）.立体培养的髓核细胞活性提高,随年龄增加细胞活性下降;指数生长期细胞分裂指数与单层培养相比有统计学意义（P＜0.05）;Ⅰ、Ⅱ型胶原含量与单层培养相比有统计学意义（P＜0.05）,A组分别与B组、C组及D组比较均有统计学意义（P＜0.05）.结论：旋转立体培养人退变椎间盘髓核细胞的活性保持良好,较单层培养能够大量、优质收集种子细胞,可用于椎间盘组织工程中种子细胞的研究.     

护士医院安全文化认知的调查研究

目的了解护士对医疗护理安全文化的认知情况,为针对性地对护士进行医院安全文化教育提供依据。方法采用经修改后的医疗安全问卷（内容包括与安全有关的病房组织文化、医院组织文化和差错事故报告系统）对233名护士进行调查。结果护士对医疗安全知识认知总分为170．58土9．63,对病房组织文化、医院组织文化、医疗差错事故报告系统的认知评分分别为78．82±6．26、43．96士5．10、57．39士5．33。不同职称、科室护士对医疗差错事故报告系统的认知水平比较,差异有统计学意义（均P〈0．05）;不同科室、学历、年龄、工龄、职称的护士对病房组织文化认知水平比较。差异无统计学意义（均P〉0．05）。不同特征（学历、科室除外）护士对医院组织文化的认知水平比较,差异有统计学意义（均P〈0．05）。结论护士对安全文化认知度低,需要加强护士医疗安全的相关意识．构建积极的医院安全文化。     

Cultured epithelial autograft (CEA) in burn treatment: three decades later

Methods for handling burn wounds have changed in recent decades and increasingly aggressive surgical approach with early tangential excision and wound closure is being applied. Split-thickness skin (STSG) autografts are the "gold standard" for burn wound closure and remain the mainstay of treatment to provide permanent wound coverage and achieve healing. In some massively burned patients, however, the burns are so extensive that donor site availability is limited. Fortunately, considerable progress has been made in the culture of human keratinocytes and it is now possible to obtain large amounts of cultured epithelium from a small skin biopsy within 3-4 weeks. Questions related to optimal cell type for culture, culture techniques, transplantation of confluent sheets or non-confluent cells, immediate and late final take, carrier and transfer modality, as well as final outcome, ability to generate an epithelium after transplantation, and scar quality are still not fully answered. Progress accomplished since Reinwald and Green first described their keratinocyte culture technique is reviewed.     

人角质形成细胞培养的实验研究

目的：探讨实验室条件下人体角质形成细胞的培养技术，为角质形成细胞的多方面研究提供可靠的细胞来源。方法：采用改良的无血清培养技术进行人体角质形成细胞的培养，用显微镜及电子显微镜观察角质形成细胞的形态特征。结果：培养的角质形成细胞在多次传代后仍保持了正常的形态。结论：用改良的培养角质形成细胞的技术可为角质形成细胞进一步的实验和临床提供可靠的、丰富的细胞来源。     

Serum-free culture of rabbit meniscal fibrochondrocytes: proliferative response

We have formulated a serum-free medium capable of supporting DNA synthesis in rabbit meniscal fibrochondrocytes at a level equivalent to 10% fetal bovine serum (FBS). The medium consists of a 1:1 mixture of Dulbecco's Modified Eagle's Medium and Ham's F-12 medium supplemented with transferrin (1 microgram/ml), selenium (1 pg/ml), trace metal mix (1:100), dexamethasone (100 ng/ml), insulin-like growth factors I and II (50 ng/ml each), pituitary fibroblast growth factor (100 ng/ml), and lactalbumin hydrolysate (2 micrograms/ml). Endothelial cell growth supplement could be substituted for lactalbumin hydrolysate to obtain similar results. Ventrex PC-1, a commercially available, low-protein, serum-free medium, was found to support proliferation of fibrochondrocytes but not as well as 10% FBS or our medium formulation. Lipid supplements, which are known to support the serum-free growth of hyaline chondrocytes, were found to be either of no value or antagonistic for the culture of fibrochondrocytes. Likewise, vitamin E alone, progesterone, putrescine, and hydrocortisone were also without benefit in our culture system. The cells had a more chondrocytic morphology when grown in defined medium as opposed to 10% FBS. The results of this study should now make it possible to identify and quantitate those factors necessary to affect meniscal repair by utilizing further techniques in vitro.     

含胎牛血清胶原酶消化法培养兔成骨细胞

目的:用含胎牛血清的胶原酶消化胎兔颅盖骨骨组织,进行体外成骨细胞培养.方法:妊娠28d的孕兔,无菌条件下剖腹取出胎兔,然后取胎兔的颅盖骨,剔净、漂洗、剪碎.用含胎牛血清的胶原酶消化,培养成骨细胞.台盼蓝染色测定细胞的存活率,细胞内碱性磷酸酶(ALP)染色鉴定和测定成骨细胞的纯度.结果:用含胎牛血清的胶原酶消化并培养出成骨细胞,台盼蓝染色拒染率高达98%,ALP染色阳性区域不少于95%.结论:用含胎牛血清胶原酶消化胎兔颅盖骨骨组织,培养出的成骨细胞具有典型的成骨细胞特征,且成分单一,存活率高.     

护士医院安全文化认知的调查研究

目的 了解护士对医疗护理安全文化的认知情况,为针对性地对护士进行医院安全文化教育提供依据.方法 采用经修改后的医疗安全问卷(内容包括与安全有关的病房组织文化、医院组织文化和差错事故报告系统)对233名护士进行调查.结果 护士对医疗安全知识认知总分为170.58±9.63,对病房组织文化、医院组织文化、医疗差错事故报告系统的认知评分分别为78.82±6.26、43.96±5.10、57.39±5.33.不同职称、科室护士对医疗差错事故报告系统的认知水平比较,差异有统计学意义(均P＜0.05);不同科室、学历、年龄、工龄、职称的护士对病房组织文化认知水平比较,差异无统计学意义(均P＞0.05).不同特征(学历、科室除外)护士对医院组织文化的认知水平比较,差异有统计学意义(均P＜0.05).结论 护士对安全文化认知度低,需要加强护士医疗安全的相关意识,构建积极的医院安全文化.     

分离肝细胞纯化培养方法的比较研究

目的 评价一种可提高肝细胞纯度和存活率的分离培养方法。方法 以体外两步胶原酶灌流法分离肝细胞,然后将其分成两组,对照组在接种培养前不经进一步处理,试验组则在应用适宜的Percoll梯度液离心纯化之后再行培养。藉台盼蓝拒染法比较两组肝细胞的存活率,采用MTT法动态比较两组肝细胞的增殖状态,在相差显微镜下观察细胞的纯度和形态。结果 未经进一步纯化处理的猪肝细胞存活率为90％±5％,鼠肝细胞存活率为80％±5％,两者纯度均约90％;经Percoll梯度液离心纯化后,其高活力肝细胞比率均提高至98％±2％,纯度可达99％以上。从开始接种到大部分肝细胞贴壁生长,试验组比对照组肝细胞的时间有所缩短。结论 用Percoll梯度液纯化新分离肝细胞,可提高肝实质细胞的活力与纯度。