DC-12 is a Bacillus strain with strong fibrinolytic activity was screened from Douchi.A novel breeding technology——whole genome shuffling was applied to enhance the production of fibrinolytic enzyme.First of all,a candidate mutant library was constructed by treating DC-12 with ultraviolet rays andhNO2,respectively.Based on studying the optimum conditions for protoplast preparation and regeneration of DC-12,multi-parental whole genome shuffling with 4 strains was conceded by protoplasts electrical fusion,then combines with the screening method of double-inactive protoplast,two strains withhigher yield of fibrinolytic enzyme were obtained effectively,which can descend stably after five generations,and their enzyme production reached 2710 IU/ml,which was increased by 4~5 fold compared with the mutants,respectively.

Objective To investigate the clinical pathological factors of papillary thyroid microcarcinoma (PTMC) with clinically node-negative sides of neck stage central lymph node metastasis.Methods Analysis of 136 cases of cNo stage of PTMC of clinical and pathological data,and using the PCR method for the detection of BRAFV600E mutation in the paraffin coated tissue.Results One hundred and thirty-six cases with PTMC lymph node metastasis rate was 38.2％ (52/136),BRAFV600E mutation rate was 44.9％ (61/136).Single factor analysis showed that the lymph node metastasis was related with BRAFV600E mutation and capsular invasion (P＜ 0.05).The size of the tumor was close to significant level (P=0.057).Multiple regression analysis showed that BRAFV600E mutation and capsular invasion were independent factors for affecting the central lymph node metastasis (P ＜ 0.05).Conclusion The routine central lymph node dissection should be performed in patients with BRAFV600E mutation and tumor invasion.

ObjectiveTo study the relation between premature and cerebral palsy(CP).MethodsTo sample all the children between the age of 1 to 6 years old for investigating CP situation in Nanning city, Heng county and Qinbei region of Qinzhou city (once named Qinzhou county ) from May to December, 1998 by qualified pediatricians.ResultsThe total investigating cases were 150,806,in which diagnosed CP were 193 cases and the incidence of CP was 1.28‰. Among them, premature infants were 872 and diagnosed CP were 46,the incidence of CP was 52.75‰ in this group. While term laboring infants were 149,934 with CP 142 and the incidence of CP was 0.95‰. The difference between two groups was very significant(P<0.001).The shorter the gestational weeks were, the higher the incidence of the CP was. Among 46 premature infants with CP, there were 18 cases' mothers(26 person times) involved in abnormal pregnancy and delivery history, accounted for 39.1%, while 36 with pregnancy complications(78.3%). There were 23 cases (30 person times) suffered from the disease during neonate, accounted for 50%.ConclusionThe premature delivery is a risk factor associated with childhood CP.

OBJECTIVE: To evaluate function and application of various biomaterials in tissue engineering repair skull defect, and to seek a reasonable skull substitutes. METHODS: Using "tissue engineering, skull defect, stem cells, cytokines, composite" as key words in Chinese, and using "tissue engineering, skull defect, mesenchymal stem cells, cytokines, materials" as key words in English, a computer-based online search was performed for publications from January 1993 to October 2009. Articles concerning biomaterials and tissue engineering skull defect repair were included. Articles describing repetitive study or Mete analysis were excluded. Function of various biomaterials in tissue engineering repair skull defect was assess in 17 articles. RESULTS: Stem cell technique is to obtain seed cells from a few myeloid tissues. Following in vitro induction and amplification, cells at a certain number could compound with stents, and construct tissue-engineered bone in vitro. The cells could sustain the biological features of osteoblasts, and fuse with bone tissue in the recipient site. Simultaneously, it could avoid the occurrence of immunological rejection. Two or over materials were compounded together, or biomaterial surface received various modification. These could promote adhesion between cells and materials, elevate cell bioactivity and maintain biological function. Following moved into seed cells using some vectors, the latter ossified in bone defect site, and secreted suitable bone growth factor, induced the differentiation of peripheral non-oriented osteogenitor cells into oriented osteogenitor cells, which could harvest adequate seed cells with osteogenic activity in a short time, and accelerate the healing of skull defects. The composite made by using three-dimensional virtual imaging and computer numerical control modeling techniques has the advantages of medical composite, precise anatomical consistency with surrounding tissues of skull defects, and perfect appearance. CONCLUSION: At present, no material can be accorded with the requirement of bone tissue engineering. Combination of some materials or modification of biomaterial surface can promote adhesion of cells and materials, elevate cell biological activity, and maintain biological function, which are presently hot focus of tissue engineering research of biomaterials.

Objective To study the effects of early mechanical strain magnitude on formation and differentiation of osteoclasts. Methods RAW 264.7 cells induced by macrophage colony-stimulating factors and osteoclast differentiation factors were subjected to 0, 1 000, 1 500, 2 000, 2 500 and 5 000 με mechanical straining for three days. The morphological changes, number of osteoclasts and proliferation of precursor cells were determined at day 7. The activity of the tartrate-reaistant acid phosphatase (TRAP) in the culture medium was detected at days 4 and 7. Results The number of osteoclasts was decreased in 2 500 με group, while it was increased in 5 000 με group. The proliferation of precursor cells was increased in 2 000 and 2 500 με group, while it was decreased significantly in 5 000 με group. There was no significant difference in the number of osteoclasts and proliferation of precursor cells among 1 000 με group, 1 500 με group and 0 με group. The activity of TRAP was decreased in 1 000, 1 500, 2 000, 2500 and 5 000 με groups at days 4 and 7 when compared with με group. Conclusions Early mechanical straining plays a direct role in formation and differentiation of osteoclasts. The high strain magnitude within physiological load inhibits osteoclast formation, while high strain magnitude beyond physiological load stimulates osteoclast formation. Low strain magnitude has nearly no impact on formation of osteoclasts. Early mechanical straining may inhibit differentiation of osteoclasts.

The study aimed to clone the open reading frame of cinnamate 4-hydroxylase (C4H) from Aquilaria sinensis and analyze the bioinformatics and expression of the gene. One unique sequence containing C4H domain was discovered in our previous reported wound transcriptome dataset of A. sinensis. The open reading frame of C4H was cloned by RT-PCR strategy with the template of mixed RNA extracted from A. sinensis stem which treated by different wound time. The bioinformatic analysis of this gene and its corresponding protein was performed. C4H expression profiles in responds to MeJA (methyl jasmonate) application were analyzed by real-time PCR. The length of C4H open reading frame (ORF) was 1 515 bp, encoding 514 amino acids. The GenBank accession number is KF134783. Inducible-experiments showed that the genes were induced by mechanical wound as well as MeJA induction, and reached the highest expression level at 8 h and 20 h, respectively. The full-length cDNA of C4H and its expression patterns will provide a foundation for further research on its function in the molecular mechanisms of aromatic compounds and flavonoids biosynthesis.
Amino Acid Sequence ; Cloning, Molecular ; Models, Molecular ; Molecular Sequence Data ; Open Reading Frames ; Oxidoreductases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Thymelaeaceae ; chemistry ; enzymology ; genetics ; Trans-Cinnamate 4-Monooxygenase ; chemistry ; genetics ; metabolism

Objective To-identify the factors associated with radiation-induced liver disease (RILD) and to describe the probability of RILD using the Lyman normal tissue complication(NTCP) model for primary liver carcinoma(PLC) treated with hypofractionated conformal therapy (CRT).Methods A total of 109 PLC patients treated with hypofractionated CRT were prospectively followed according to the Child-Pugh classification for liver cirrhosis,93 patients in class A and 16 in class B.The mean dose of radi- ation to the isocenter was (53.5?5.5) Gy,fractions of (4.8?0.5) Gy,with interfraction interval of 48 hours and irradiation 3 times per week.Maximal likelihood analysis yielded the best estimates of parameters of the Lyman NTCP model for all patients;Child-Pugh A and Child-Pugh B patients,respectively.Results Of all the patients,17 developed RILD (17/109),8 in Child-Pugh A(8/93 ) and 9 in Child-Pugh B(9/ 16).By multivariate analysis,only the Child-Pugh Grade of liver cirrhosis was the independent factor (P= 0.000) associated with the developing of RILD.The best estimates of the NTCP parameters for all 109 pa- tients were n=1.1,m=0.35 and TD_(50) (1)=38.5 Gy.The n,m,TD_(50) (1) estimated from patients with Child-Pugh A was 1.1,0.28,40.5 Gy,respectively,compared with 0.7,0.43,23 Gy respectively,for patients with Child-Pugh B.Conclusions Primary liver cancer patients who possess Child-Pugh B cirrho- sis would present a significantly greater susceptibility to RILD after hypofractionated CRT than patients with Child-Pugh A cirrhosis.The predominant risk factor for developing RILD is the severity of hepatic cirrhosis in the liver of PLC patients.

BACKGROUND: Seed cell exerting its function is required to depend on the extracellular matrix in tissue engineering, so that biocompatible material is important to be selected. OBJECTIVE: To prepare a novel composite scaffolds of chitosan/collagen/nano-hydroxyapatite (HA-CS/Col/nHAP) and to optimize the technology of tissue engineered-stents according to the circumstances of cell adhesion.METHODS: Chitosan was modified by hyaluronate acid. The structure was observed by differential scanning calorimetry and the Fourier transformed infrared spectroscopy. Three composites of HA-CS/Col/nHAP according to different ratio of chitosan and collagen solution (1: 2; 1: 1 and 2: 1) were prepared. The composite scaffolds were co-cultured with osteoblast MC3T3-E1, and the proliferation and cell growth curve were measured by CCK-8 method. RESULTS AND CONCLUSION: Hyaluronic acid and chitosan were crosslinked with amide linkage. Pore size was on the range from 50 μm to 250 μm. Porosity was increased with increased collagen level and elastic modulus, but density was reduced. Increased collagen content was beneficial for cell adhesion and proliferation on stent in the primary phase of cell co-culture. However, from day 10, no significant difference was determined among three samples. At the beginning of cell culture, cells adhered to the airspace insides the composite scaffolds. In the following days, cells grew in a colony manner, and cell-cell junction could be easily observed. These indicate that HA-CS/Col /nHAP composite scaffolds can improve the adhesion and proliferation of osteoblast. The ratio of chitosan to collagen volume at 1: 1 was optimal.

Objective To explore the effects of H2S on cerebral injury after cardiopulmonary resuscitation (CPR) and its mechanism.Methods Forty-five healthy Sprague-Dawley (SD) rats were randomly (random number) divided into shame-operated group ( group A,n =5 ),resuscitation group ( group B,further divided into four subgroups as per rats sacrificed 6 h,12 h,24 h,and 72 h after resuscitation,n =5),and NaHS pretreatment group ( group C,further divided into 4 subgroups as done in group B).The ratio of water content in brain tissue was calculated.The content of H2S in cerebral cortex of rats in all groups was determined by using universal microplate reader. Immunohistochemistry method was used to count the Nestin-positive cells. Results The content of H2S in hippocampus area of brain showed dramatic changes from rising up at first and then to lowering down to the minimum and finally returning to the original level in 72 h in B group.Compare to group B,brain water content was lesser ( P ＜0.05 or P ＜ 0.01 ) and the levels of Nestin in hippocampus increased in group C(P＜0.05 or P ＜0.01).The neurological deficit score (NDS) was improved (P ＜0.05 or P ＜0.01) and pathological changes in hippocampus of rat brain detected by using hemotoxylin - eosin staining were slighter in group C in comparison with group B.Conclusions Endogenous H2S may involve in the course of formation and progress of cerebral injury after CPR and small dose of NaHS (exogenous H2S) can improve NDS by decreasing cerebral edema and up-regulating Nestin level in hippocampus of brain,playing a protection role in cerebral injury after CPR.

This study aimed to investigate the effects of Shiquan Dabu Tang (SDT) on growth and angiogenesis of subcutaneously implanted tumors, hepatic metastases, and incision-implanted tumors after surgical removal of primary colon tumor in mice.