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.

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.

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 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.

[ ABSTRACT] AIM:To investigate the genetic cause of 2 Chinese families with Marfan syndrome .METHODS:The clinical and laboratory investigations were performed in the 2 unrelated Chinese families .Family 1 had 1 patient with cardiac problem.Family 2 had 2 patients:one died, and the other with respiratory and cardiac problems .Next generation sequencing and Sanger sequencing in the Marfan syndrome causal gene FBN1 were performed in the patient , his unaffected sister and the parents of family 1.Sanger sequencing covering all the exons and intron-exon boundaries were performed in the patient and the parents in family 2.Bioinformatic analysis was engaged in the variations unravelled .Fifty healthy indi-viduals were also investigated in the same manner .RESULTS:Both patients were diagnosed with Marfan syndrome .A no-vel mutation c.4685G>A (p.Cys1562Tyr) was detected in the patient of family 1 but was absent in his parents and the unaffected sister .This is a previously unreported novel mutation .In the mutation a conserved Cys was substituted by a Tyr in amino acid 1562 affecting a TGF-βbinding domain and the secondary structure in the encoded protein .We also detected the mutation c.3706T>C (p.Cys1236Arg) in the patient of family 2.It was absent in the unaffected parents , and there-fore was a de novo mutation too.This mutation has been previously reported and known to be associated with neonatal Marfan syndrome .Both mutations were absent in the 50 healthy controls .We also compared the genotype and phenotypes of the 2 families.CONCLUSION:We report 2 de novo mutations in 2 Chinese families with Marfan syndrome .One of the 2 mutations is novel.The phenotype of the mutation c.4685G>A(p.Cys1562Tyr) in family 1 is associated with classical Marfan syndrome, while that of c.3706T>C (p.Cys1236Arg) in family 2 is with neonatal type of Marfan syndrome .De novo mutations may be a cause for a proportion of mutations underlying the disease .The novel mutation also expends the mutational spectrum of the FBN1 gene.

Objective To investigate the effect of video-assisted thoracoscopic surgery( VATS) in thoracic disease,and the feasibility to carry out VATS for basic hospital. Methods The data of VATS treatment were collected to compare the differences between study group and control group,and evaluate the the feasibility to carry out VATS for basic hospital. Results The operation time was (100. 75±22. 72) min, blood loss was (54. 27±26. 21) mL,postoperative drainage was (920. 67±171. 99) mL. The postoperative complications were fewer,post-operative hospital stay was shorter,incision time was shorter(P=0. 000) and pain scores was lower(P=0. 000) in study group than that in control group. Basic hospital has the capacity to conduct this technical. Conclusion VATS is feasible to carry out in basic hospital.

Objective To discuss the value of 16-slice spiral CT urography in clinical diagnosis.Methods To compare the images between CR excretory urography,CT and MPR,which are the results of 26 cases who were examined by both CR and 16-slice spiral CT urography.Results From the comparison,it is found that the spiral CT urography images are better than that of the CR excertory urography,and at the same time,the relationship between the parenchyma of the renal,renal pelvis,ureter,bladder and the urinary organs and adjacent structures can be demonstrated clearly.Conclusion The scanning time of the spiral CT is shorter.The machine can not only reconstruct powerfully but also work out better images,and the result is better than CR excretory urography which can not detect some details in its images.Therefore CR excretory urogaphy can be replaced by 16-slice spiral CT urography regardless of its cost.