Microbial lipase,one of important industrial biocatalysts,has been used widely in many industrial and agricultural fields.It is always the research focus to screen,mine and develop the microbial lipases with novel catalytic activity and high stability.This paper introduces briefly the pathways and methods to mine novel microbial lipase resources from six aspects,including extremophile,metagenome,genome database,protein engineering,immobilization,chemical modification,etc.

Microbial lipases are important industrial biocatalysts with the character of stereoselectivity,site selectivity and high catalytic activity with few side effects.They have been used widely in many industrial and agricultural fields.The technology of protein engineering has been successfully applied to improve the activity and stability of microbial lipases,which will raise the competitive capacity of microbial lipase preparations and enlarge theirs application fields.The strategies,the problems and the prospects of protein engineering technology which have been applied to modify the microbial lipases was surveied.

AIM:To investigate the clinical effect of anti-vascular endothelial growth factor (VEGF) drugs combined with laser photocoagulation for diabetic macular edema (DME).METHODS: Totally 94 patients (141 eyes) with DME from June to December 2015 in our hospital were selected and randomly divided into combined group of 47 cases (68 eyes, ranibizumab combined with laser therapy) and the control group of 47 cases (73 eyes, laser treatment).The levels of best corrected visual acuity (BCVA), macular central retinal thickness (CRT), total macular volume (TMV) and macular edema level were compared between the two groups at different time after treatment.RESULTS: The mean values of BCVA in the combined group were higher than those in the control group at 2, 6 and 12wk, and the difference was statistically significant (P<0.05).At 2, 6 and 12wk after treatment, the CRT and TMV values of the combined group were lower than those of the control group, the difference was statistically significant (P<0.05).After treated for 12wk, patients with macular edema of combined group was 80.9% in mild level, 17.7% in moderate level, 1.5% in severe level, those of the control group was 60.0%, 31.5%, 5.5%, the difference between the two groups was statistically significant (P<0.05).CONCLUSION: The effect of anti-VEGF drugs combined with laser therapy for DME patients is better than that of single laser therapy alone.

Objective:To stablish of chemiluminescent Southern blot detection system for examining HBV DNA replication intermediates in HepG2.2.15,and analyse the inhibition of HBV replication with three kind of drugs with different targets.Methods:The HBV DNA replication intermediates were extracted and analyzed by Southern blot with HBV probe,which(pTHBV1047) was labelling with digoxigenin.The results of the hybridization were detected by chemiluminescent,and the condition of hybridization was optimized.After treated with lamivudine,Bay41-4109,?-Galcer in different concentration,the HBV DNA from the HepG2215 cells were detected with the system.Results:the sensitivity of the system was 1pg of pTHBV1047,and HBV specific positive signals was detected with the DNA from HepG2.2.15.The three kinds of drugs can inhibit the HBV replication obviously with chemiluminescent Southern blot detection system,the IC50 were 1.53?mol/L,0.41?mol/L,0.01?mol/L.Conclusion:The HBV replication intermediates from the cell of HepG2.2.15 can reflect the antiviral effect accurately with different targets drugs and this mothod would be used in the study of Chinese-midicine.

Animals ; Animals, Newborn ; Hypoxia-Ischemia, Brain ; metabolism ; Neuroprotective Agents ; pharmacology ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; Progesterone ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVETo investigate the expressions of nestin and vascular endothehal growth factor (VEGF) mRNAs in rat brain tissue after cerebral ischemia-reperfusion injury and their changes in response to Tongxinluo (a traditional Chinese herbal preparation) treatment.

METHODSCerebral ischemia was induced in rats by temporary middle cerebral artery occlusion (MCAO) followed by treatment with Tongxinluo at high and low doses. On days 3, 7, 14 and 21 after MCAO, nestin and VEGF mRNA expressions in the ependyma, subventricular zone (SVZ), and hippocampal subdentate gyrus zone (HDG) in the ischemic hemisphere were quantitatively analyzed using immunohistochemistry and RT-PCR.

RESULTSCompared with the sham-operated group, the rats with cerebral ischemia-reperfusion injury showed significantly increased nestin-positive neurons and VEGF mRNA expression in the SVZ and HDG 7, 14 and 21 days after MCAO (P<0.05). Treatment with Tongxinluo, especially at high doses, significantly increased the number of nestin-positive neurons and VEGF mRNA expression in the rats 7, 14 and 21 days after MCAO (P<0.05).

CONCLUSIONFocal cerebral ischemia in rats results in rapid response and proliferation of neural stem cells in the SVZ and HDG in the ischemic hemisphere possibly by increasing VEGF mRNA expression in the adjacent tissues around the ischemic focus. Tongxinluo may enhance the differentiation and proliferation capacity of the neural stem cells after MCAO by inducing the expression of VEGF mRNA.

Animals ; Brain Ischemia ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Intermediate Filament Proteins ; genetics ; metabolism ; Male ; Nerve Tissue Proteins ; genetics ; metabolism ; Nestin ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Objective To investigate the phenotypes and genotypes of a hereditary protein C(PC) deficiency pedigree.Methods Imrnunoassay(ELISA)was used for PC antigen and PS antigen; Immunoturbidimetry assay was used for measuring AT antigen;Chromogenic substrate assay was used for measuring the activity of PC,PS and AT in Sysmex 1500 automatic Blood Coagulation Analyzer.Polymerase chain reaction(PCR)for amplification of the fragment of each exon and side sequences of PC gene in 10 members of the 3 generations;Direct DNA sequencing was used to examine the mutation site.Results Among 10 members of the 3 generation pedigree,8 of them had a PC:Ag level of 1.06-1.92 mg/L(normal references 3.00-6.00 rag/L),the activity of PC was between 41% and 67%(normal references 70%- 140%),which was significantly lower than the normal references while the levels of PS:Ag,PS:A,AT:Ag and AT:A were all within normal range.DNA sequencing analysis showed that there was a G to T mutation in exon IX of the PC gene at 12 918 position in 8 members.This mutation resulted in the substitution of terminator TGA for TGG which encoding tryptophan at 372 amino acid.There was a polymorphism in 2 405C/ T,2 418A/G,2 583A/T in the promotor area.Conclusions This pedigree is a type I hereditary protein C deficiency.There is a G12 918T mutation in exon IX of PC gene.This mutation is reported for the first time and there is a polymorphism in 2 405C/T,2 418A/G,2 583A/T in the promotor area.

OBJECTIVETo study the efficacy and complications of the method for reduction mammaplasty in patients with breast benign diseases.

METHODSFrom November 1980 to December 2001, reduction mammaplasty was performed in 27 patients with breast hypertrophy, ptosis and benign diseases. The operation methods were selected according to the characters of the diseases and the extent of breast hypertrophy and ptosis. 9 patients received reduction mammaplasty using an inferior pyramidal pedicle technique; 16 patients received Mckissock vertical bipedicle technique and 2 patients received the bicyclic incision technique.

RESULTSThe successful rate was 94.2%. Three breasts developed areola necrosis in Mckissock. Symptoms caused by breast hypertrophy and benign diseases were improved apparently.

CONCLUSIONReduction mammaplasty is the best option for the treatment of breast hypertrophy and ptosis with benign diseases. The method of reduction mammaplasty should be taken individually.

Aged ; Breast ; pathology ; surgery ; Breast Diseases ; surgery ; Female ; Humans ; Hypertrophy ; surgery ; Mammaplasty ; methods

OBJECTIVETo evaluate the possibility that using intracoronary delivery of autologus bone marrow-derived mesenchymal stem cells (MSCs) to improve the cardiac function after acute myocardial infarction (AMI) in miniature pig.

METHODSMSCs were cultured in Dulbecco's modified Eagle's medium-F12 (DMEM/F12) medium. AMI model was made by blocking the blood stream of the first diagonal branch in miniature pig, and released the branch after 90 minutes. After 10-14 days, (4-6) x 10(7) culture-expanded autologus 4', 6-diamidino-2-phenylindole (DAPI)-labelled MSCs were transplanted into each host heart's AMI area through intracoronary way. Ultrasonic cardiography (UCG) was performed to observe the left ventricular function at 3 months after transplantation. The cellular transplanted hearts were harvested and investigated by immunohistochemical analysis.

RESULTSLeft ventricular function of the MSCs group was improved significantly 3 months later compared with the control group [(54.65 +/- 3.39) vs (43.98 +/- 4.21)%, (P < 0.01)]. Exogenous MSCs survived and site-differentiated into cardiomyocytes in infracted hearts.

CONCLUSIONMSCs can play a benificial role to repair damaged heart. Heart function can be improved after MSCs transplantation in porcine myocardial infarction model.

Animals ; Female ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Myocardial Infarction ; pathology ; physiopathology ; therapy ; Swine ; Swine, Miniature ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVEReview and discuss anesthesia and operative approach of resection of the trachea for different tracheal diseases, especially for the resection of long-segment of trachea. At the same time to introduce the method of reconstruction of long-segment of trachea with Zhao's (two-stage procedure with memory-alloy mesh) artificial trachea.

METHODSRetrospective study of 18 cases of tracheal resection, analysis of the relation between the choice of anesthetic and operative approach.

RESULTSGeneral anesthesia through cut open the trachea with local anesthesia in 2 cases, general anesthesia through previous tracheotomy in 2 cases, extracorporeal circulation in 2 cases, general anesthesia through endotracheal tube in 12 cases. There were no anesthetic or operative death. Local resection in 3 cases, segmental resection in 15 cases. The longest segmental resection was 8.0 cm. Primary anastomosis after segmental resection in 8 cases, Reconstruction with Zhao's artificial trachea in 7 cases. Postoperative follow-up was 5 months to 8 years. Four cases died from systemic metastasis or other reasons at 4, 11 and 12 months, respectively.

CONCLUSIONSDifferent methods of anesthetic and operative procedures should be used for different patients. Extracorporeal circulation used for patient with highest dangerous condition, or, for which could be inserted endotracheal tube by tracheotomy with local anesthesia. Conservative local resection performed only for patients with very bad general condition. Segmental resection less than 5 cm long could be reconstructed with primary reanastomosis. Resection longer than 5.5 cm could be reconstructed with Zhao's artificial trachea.

Adolescent ; Adult ; Aged ; Anesthesia, General ; methods ; Anesthesia, Local ; Artificial Organs ; Extracorporeal Circulation ; Female ; Humans ; Intubation, Intratracheal ; Male ; Middle Aged ; Prosthesis Implantation ; Retrospective Studies ; Thoracotomy ; Tracheal Diseases ; surgery ; Tracheotomy ; methods