The metagenomic DNA was extracted from the deep sea sediment with the depth of 900m of Prydz Bay, Antarctic. A lipase gene (lip3) with the size of 948bp was cloned from the metagenomic DNA by PCR with the primers designed. The deduced Lip3 protein was composed of 315 amino acids (AA) with a molecular mass of 34.577 ku. The motifs GFGNS(GXGXS)and G-N-S-M-G(GXSXG)in the AA sequences of Lip3 were found to be conserved in other lipase. They were most conserved sequence among the serine hydrolase and were necessary for the activity. A 35 ku of Lip3 was purified by Ni-NTA chelating sepharose column from the extract of recombinant E.coli Top 10F′ cell harboring a pLLP-OmpA plasmid inserted with lip3. The purified Lip3 was most active at 25℃ and kept 22% of activity at 0℃. Only 10% of activity was retained after it was incubated at 35℃ for 60 min. The optimal pH value for the Lip3 activity was 8.0. The Km value of the enzyme towards p-nitrophenyl palmitate increased with the increasing of assayed temperature. These results indicated that Lip3 was a typical alkaline cold active enzyme.

Objective To establish a mouse model of early lung adenocarcinoma to serve the imaging studies of early lung adenocarcinoma .Methods Two-hundred and ten 4-week old SPF female Kunming mice were randomly divided into 5 experimental groups (40 mice per group) and 1 control group (10 mice).The mice of experimental groups were subcutaneously injected with a dose of 0.2 ml 1-methyl-3-nitro-1-nitroso-guanidine ( MNNG) solution in concentration of 2.0 mg/mL weekly for 1, 2, 4, 8, 12 weeks (group A-E), respectively, while the mice of control group was subcutane-ously injected with 0.2 mL saline weekly for 12 weeks.Ten mice were randomly sacrificed from each experimental group at the 60th, 80th, 100th and 120th days after initial injection ( group A60 , group A80 , group A100 , group A120; group B60 ,……and so on) , and the bilateral lungs were dissected .The tumor occurrence rate , number and size of lung tumors were observedandrecorded.Results 1.Thetumoroccurrencerateandnumberoflungtumorswerepositivelyproportionalto the time and dose of MNNG injection .No tumor was found in any group at the 60th day.2.At the 80th day, the tumor oc-currence rate in the groups A, B, C, D and E was 0, 10%, 30%, 40%and 50%, respectively.The number of tumors in each group was 0, 4, 42, 60 and 81, respectively.The number of tumors smaller than 0.5 mm in diameter was 0, 4, 25, 31 and 40, respectively.3.At the 100th day, the tumor occurrence rate was 20%, 40%, 100%, 100%and 100%, re-spectively.The number of tumors was 6, 19, 187, 223 and 301, respectively, and the number of tumors smaller than 0.5 mm in diameter was 5, 16, 132, 124 and 123, respectively.4.At the 120th day, the tumor occurrence rate was 30%, 50%, 100%, 100%and 100%, respectively.The number of tumors was 30, 124, 302, 351 and 362 per group, respec-tively.The number of tumors smaller than 0.5 mm in diameter was 21, 98, 123, 140 and 108, respectively.The induced lung adenocarcinomas were confirmed by pathology .Conclusions MNNG solution 0.2 mL (2.0 mg/mL) subcutaneously injected weekly for 4 weeks can produce 100%occurrence rate of lung adenocarcinoma in mice and most tumors are smaller than 0.5 mm in diameter at 100th day after initial injection .It is a most suitable model for imaging studies of mouse early lung adenocarcinoma .

Objective To detect the genotypes of human papillomaviral infection(HPV)in 970 women who were suspicious as HPV infection, and to investigate its clinical significance. Methods Cervical cast-off cells were acquired with the special cervix brush. DNA hybridization technique was applied to detecting HPV genotypes in 970 clinical specimens. Results Totally 390 case(40.2%) were HPV positive among 970 clinical specimens. A total of 12 subtypes were detected. In 390 cases of HPV infection,260 cases (26.8%) were infected with single HPV subtype, and while 130 cases(13.4%) were with infection of two or above HPV subtypes. HPV6 and 11 were the most frequent low-risk sub-types,and in the high-risk subtypes,HPV16,33,52,58 and 18 were dominant. Conclusion The cervix HPV infection rate is considerable in women in Wuxi area. HPV6 and 11 are the most frequent sub-types. The determination of HPV genotypes plays all important role in early discovery, prevention and treatment of gential tract and cervical cancer.

Objective To research and develop a model of the representative active component's content by NIR spectroscopy,so as to realize the on-line quality control of extracting process for multiple herbal medicine system in product scale.Methods The on-line NIR detection of extracting process was used to obtain the NIR spectrum,HPLC detection of the extracts was carried out to determine the content of danshensu,and PLS method was used to establish the relationship between the information of NIR and HPLC.Results The optimum NIR wavelength range was 9 715-7 082 cm-1,R=0.959 4,RMSEC=0.049 4,the average relative error was 7.2%.Conclusion NIR Technique could be used in the on-line quality control of Compound Danshen's extracting process.

The triple negative breast cancer is one of importance in clinical subtypes of breast cancer, which is easy to recur and metastasis, and its prognosis is very poor.Radiotherapy, as an effective method for breast cancer,can reduce the risk of local recurrence.This article elaborates its characteristics,the progress of ra-diotherapy in the breast conserving surgery and modified radical mastectomy in triple negative breast cancer,when is the appropriate time for radiotherapy and radiotherapy sensitization,and hope that it will be helpful to the treat-ments.

Objective:To develop lipiodol-containing calcium alginate microspheres (LAMs) for embolization,and study the characterization for emoblization and the radiopacity. Methods:LAMs were prepared by dripping method. The preparation of LAMs was optimized by orthogonal experiment which involved effects of three factors (the volume ratio of lipiodol to the external aqueous solution,airflow rate,and the weight pushing the injector) at three levels on the responses to the size,polydisperse index and entrapment efficiency of LAMs. The morphology of LAMs was observed under microscope. The elasticity of LAMs was investigated by texture analyzer. The capability injected through catheter of LAMs was monitored by video spinning-drop tensionmeter. The radiopacity of LAMs was measured by X-ray imaging system after LAMs were injected into vas of a rat. Results:The optimal condition for preparation of LAMs was:the volume ratio of lipiodol to the external aqueous solution was 3∶ 10,airflow rate was 40 g/mL and the weight pushing the injector was 100 g. According to the optimized condition,LAMs were prepared and characterized. The mean diameter of LAMs was (493.9?42.6) ?m,the polydisperse index was 1.02 and the entrapment efficiency was (88.97?1.09)%. The LAMs were with round shape and smooth surface in view of photograph of microscope. The maximum average load was (1.09?0.18) N when LAMs were compressed to 60%. The LAMs were injected through catheter without much difficulty. The radiopacity of LAMs in rats was demonstrated to be visible under X-ray photography system. Conclusion:The radiopaque LAMs developed are suitable for the arterial embolization,with round shape,proper size,good elasticity,easy handling character and visible property under X-ray imaging. The radiopaque embolic agent is supposed to be useful for emoblization therapy.

Objective The volatile oil was further purified to get the patchouli alcohol by molecular distillation taking patchouli oil which contained 23.97% of patchouli alcohol as raw material.Methods The best process parameters,including temperature,feed rate,and scraping rate,of molecular distillation technique for separating patchouli oil were studied by orthogonal test.The content was detected by GC under the conditions of elastic quartz capillary column HP-5,detector temperature 280 ℃,gasification temperature 280 ℃,distributary refer 20∶1.The effects of evaporation temperature on content and extraction rate of the patchouli oil had been investigated and the difference between the molecular and vapor distillations was studied.Results Content and extraction rate of patchouli alcohol under the best evaporation temperature 65 ℃ by molecular distillation reached to 40.71% and 76.55%,respectively.Conclusion The molecular distillation technique that could get the product with higher purity and extracting rate than that by vapor distillation in patchouli oil purification has a very good prospect in industry.

Objective To optimize the enzymolysis technology of ginsenoside Rg1.Methods Taking ginsenoside Rg1 content as the index,orthogonal design method was used for optimization and HPLC for determination.Results Cellulase enzymolysis was the best extracting process,and enzyme amount,enzymolysis time,and enzymolysis temperature had obvious effect on the extraction of ginsenoside Rg1.The optimum extraction technologies were as follows: cellulase amount was 1.4%,enzymolysis time 60 min,the enzymolysis temperature 45 ℃.Conclusion The optimization extraction technology is simple,steady,and the extracting rate is high.

An adult patient with hypophosphatasia caused by compound heterozygous mutations in alkaline phosphatase,liver /bone /kidney(ALPL)gene was investigated through comprehensively reviewing the medical history and clinical records of the proband and her family members in order to better understand the disease.The proband and her older sister had mild decreased serum alkaline phosphatase level accompanied with frequently nontraumatic fractures at limbs and all the teeth fell off at the age of 20 and 7, respectively.Both of them carried a missense mutation c.407G>A(p.Arg136His)in exon 5 and a deletion mutation c.1318_1320delAAC(p.Asn440del)in exon 12 simultaneously.Other four family members were p.Arg136His mutation carriers and two members were p.Asn440del mutation carriers.We found that p.Asn440del mutation was associated with the oral disorders.In this family, compound heterozygous manifested more serious symptoms, while heterozygous showed relatively mild symptoms.In addition, it is necessary to differentiate it from primary osteoporosis and other diseases of disturbed bone mineralization.