Objective:To study the expression of CD137L in laryngeal carcinoma, and to analyze its clinicopathological significance.Method:The expression of CD137L in 50 laryngeal carcinoma specimens and 9 normal laryngeal mucous tissues were detected by immunohistochemical staining.Result:The positive CD137L staining were found in all 50 cases of laryngeal carcinomas (100%), while its staining were negative in normal laryngeal mucous. There was significant difference between two groups(P<0.01). The positive ratio of CD137L staining had no relationship with the factors such as age, sex and tumor site, while it had significant correlation with the pathological stage,T stage and lymph node metastasis.Conclusion:The expression of CD137L might play an important role in the development of laryngeal carcinomas.

BACKGROUND:Bone marrow mesenchymal stem cellcontent is less under normal conditions, and easily confounded with other cells. Therefore, to establish a simple feasible in vitro cultured amplification method and to obtain a large number of stable bone marrow mesenchymal stem cells are of important theoretical significance and application value. OBJECTIVE:To establish a simple feasible in vitro culture and amplification method and to obtain numerous stable bone marrow mesenchymal stem cells. METHODS:Bone marrow mesenchymal stem cells were isolated from Sprague-Dawley rats and cultured in vitro using density gradient centrifugation and adherence separation screening. cellmorphological changes were observed under an inverted light microscope. Submicroscopic structure of cells was observed under a transmission electron microscope. Living cellnumber was counted using Trypan blue exclusion method. cellgrowth curve was drawn. cellcycle was analyzed using flow cytometry. The expression of c-kit and CD45 was measured using immunocytochemistry. CD45 expression was analyzed using flow cytometry. RESULTS AND CONCLUSION:(1) celladhesion and pseudopodia could be seen under an inverted light microscope at 24 hours after inoculation;cellcolony formed at 4 days;90%cellconfluence was found at 14 days. After passage, the cells were tended to homogenous and became fibroblast-like cells, which showed whirl-like growing or flamboyancy growing. (2) The size of passage 3 bone marrow mesenchymal stem cells was smal and nucleolus was large, with clear nucleoli under a transmission electron microscope, with the presence of sparse chromatin and low electron density. There was microvil us on the surface of cells, abundant ribosomes could be seen in the cytoplasm with few other cellular organs such as endoplasmic reticulum, mitochondria and Golgi complex. These showed that ultrastructural structure had undifferentiated features. (3) The number of passage 3 bone marrow mesenchymal stem cells was reduced at 1 day after inoculation. The cells began to grow at 2 days, and entered the period of exponential growth at 3 days, entered the flat period at 7 days and the number of cells began to decrease at 9 days. Growth curve exhibited“S”shape. (4) The percentage of S phase cells was 21.1%as detected by flow cytometry after passage 3 bone marrow mesenchymal stem cells were stained by DNA. (5) Immunocytochemistry and flow cytometry had proved that the c-kit positive rate of cells was 53.3%and CD45 positive rate was 1.68%. (6) After osteogenic induction of mesenchymal stem cells for 16 days, cells became oval, and short processes connected each other. The cytoplasm was dark, suggesting that it may be rich in rough endoplasmic reticulum and Golgi complex. They can secrete osteoid. Results verified that obtained bone marrow mesenchymal stem cells stably grew, and actively proliferated.

Vectors used to carry foreign genes play an important role in gene therapy, among which, the adeno-associated virus (AAV) has many advantages, such as nonpathogenicity, low immunogenicity, stable and long-term expression and multiple-tissue-type infection, etc. These advantages have made AAV one of the most potential vectors in gene therapy, and widely used in many clinical researches, for example, Parkinson's disease. This paper introduces the biological characteristics of AAV and the latest research progress of AAV carrying neurotrophic factor, dopamine synthesis related enzymes and glutamic acid decarboxylase gene in the gene therapy of Parkinson's disease.

OBJECTIVE:To provide reference for developing the implementation path of patent standardization for pharmaceuti-cal industry. METHODS:For the situation that pharmaceutical industry is still in the exploratory stage and lacks of viable experi-ence,the mature experiences of electronic communication industry in patent leading standardization implementation,authoritative organizations helping implementing patent standardization and industry alliance to promote the development of patent standardiza-tion were used as reference,combining with standard development cycle,trying to develop the implementation path of patent stan-dardization that met characteristics of pharmaceutical industry. RESULTS & CONCLUSIONS:The implementation path of patent standardization for pharmaceutical industry was divided into standard formation stage,promotion stage and upgrade stage. The main path in each stage was from patent to registration standards officially formed,guiding by policies and regulations of Food and Drug Administration to conduct standard promote by drug generation,and upgrading the drug standard by patent replacement.

BACKGROUND: Fibrin is a natural biodegradable polymer material with good histocompatibility. Basic fibroblast growth factor (bFGF) is an important polypeptide growth factor with mitogenic effect. Fibrin gel (FG) combined with bFGF is conducive to cell migration and promotes proliferation and differentiation. OBJECTIVE: To investigate the effects of fibrin gel combined with bFGF on alkaline phosphatase activity (ALP) in fetal rat limb cells. METHODS: Four groups were included: FG group: fetal rat limb cells were seeded in FG (three-dimensional, 3D) with normal medium; FG + bFGF group: cells were seeded in FG containing 10 μg/L bFGF for 24-hour 3D culture; bFGF group: cells were seeded in 10 μg/L bFGF medium; normal control group: cells were seeded in normal culture medium. Under aseptic condition, rat fetal limb cells were isolated. Passage 3 cells were used for each group above mentioned. ALP activity, mRNA expression and cell morphology were analyzed at 3, 5, 7, 14, 21, and 28 days after culture. RESULTS AND CONCLUSION: The cells in the FG had more processes and formed an interconnected network after 7 days of incubation, while the cells in no gels remained cuboidal or "spindle-shaped". At 7, 14, and 21 days, ALP activity was greater in the FG+ bFGF group than in the other groups (P < 0.05). At each time point, mRNA expression of ALP was greater in the FG+ bFGF group than in the control group (P < 0.01). These findings indicate that FG combined with bFGF is conducive to cell morphogenesis and markedly enhances ALP activity.

Objective To prepare the supersaturation self-emulsifying drug delivery system(S-SEDDS) containing silymarin and to evaluate its basic properties.Methods With the time of self-emulsifying,the consequence of color visual examination and particle size as parameters,the optimum formulations of silymarin SSEDDS were screened by solubility test,compatibility tests and pseudo ternary phase diagrams.The silymarin concentration was determined by HPLC.The in vitro dissolution characteristics of silymarin S-SEDDS were investigated with silymarin SEDDS as control.Results The optimum silymarin S-SEDDS was composed of medium chain triglycerides(MCT) 40%,Cremophor RH40(ethoxylated hydrogenatedcastor oil) 48%,Labrasol 12%.The time of self-emulsifying was less than 3 min,the average particle diameter was 49.6 nm,the adding amount of hydroxypropyl methylcellulose(HPMC) was 50 mg/g,and the average content of silymarin was 39.3 mg/g.The in vitro dissolution test of silymarin S-SEDDS showed that the presence of a small amount of cellulosic polymer effectively sustained a metastable supersaturated state by retarding precipitation kinetics.Conclusion The designed formulation of silymarin S-SEDDS is reasonable and provides a strong foundation for further development of new preparations.

BACKGROUND: Fibrin is a kind of high polymer materials with biodegradation and good histocompatibility, and is a vector that can promote celland exogenous growth factor release. Fibrin stabilizing factor XIII has been verified to contribute to the migration of undifferentiated mesenchymal stem cels in gel scaffold with high crosslinking, and promote cellproliferation and differentiation. OBJECTIVE:To observe rat mesenchymal stem cellbehavior in a fibrin gel. METHODS:The rat fetal limbs cels was separated under the aseptic condition. The passage 3 cels were seeded in 0, 5, 10 and 20 g/L fibrin gel. cellmorphology was observed by inverted phase microscope and laser scanning confocal microscopy. Alkaline phosphatase activity and calcium deposition were measured respectively using a microplate reader and von Kossa staining. RESULTS AND CONCLUSION: 5 g/L fibrin gel contributed to cellmorphological changes, and 20 g/L fibrin gel contributed to osteogenic differentiation. Compared with the control group, alkaline phosphatase activity was higher in the formulations containing a 20 g/L fibrinogen concentration. Smal mineralization nodules were observed at 21 and 28 days in a formulation containing both 10 and 20 g/L fibrinogen concentration, but no mineralization was detected in the control group. These results indicate that morphology and osteogenic differentiation of rat mesenchymal stem cels depended on the fibrinogen concentration, suggesting that fibrin gel is conducive to osteogenic differentiation of mesenchymal stem cels.

Purpose To clarify the morphological parameter and describe the distance from the insertion of the lateral ankle ligaments to the adjacent bony landmarks through precisely anatomical explore of human cadaveric ankles,so as to provide anatomical evidences for the reconstruction of lateral ankle ligaments.Methods Nineteen ankle specimens were dissected to isolate the lateral ankle ligaments and measure the morphological parameters such as length,width,thickness and the distance from the insertion of the lateral ankle ligaments to the adjacent bony landmarks.Results The average length of anterior talofibular ligaments (ATFL) was 23.1 ± 2.98 mm,among which 8 were single-banded(42.1％)and 11 were double-banded(57.9％).The average distance from the fibular origination of ATFL to the anterior tubercle of fibula(AA)was 17.1 ± 3.00 mm,to the fibular obscure tubercle(AO)was 5.1 ± 1.69 mm,to the tip of the fibula(AT)was 14.1 ± 2.86 mm.The distances from the talus insertion of ATFL to the superior and inferior talus articular surface were 11.4 ± 2.25 mm and 18.4 ± 2.30 mm respectively,to the anterior lateral talus chondral surface was 4.8 ± 1.42 mm.The average length of calcaneofibular ligament(CFL)was 31.4 ± 3.55 mm.The average distance of the fibular origination from ATFL to CFL was 6.4 ± 2.55 mm.The average angle between ATFL and CFL was 116.6 ± 12.69°.The distance from the calcaneus insertion of CFL to the peroneal tubercle(CP)was 15.4 ± 2.86 mm,to the posterior superior border of calcaneus(CC)was 13.9 ± 2.46 mm,to the subtalar joint surface was 15.2 ± 3.21 mm.The coefficient variation assessing the anatomical reliability of different bony landmarks were as follows:ATFL fibular origination AA(17.54％) ＜AT(20.28％) ＜ AO(33.14％),CFL calcaneus insertion CC(17.70％)＜CP(18.57％)＜CS(21.1％).Conclusion Certain variations exist in the morphological parameters and the distances from the insertion of the lateral ankle ligaments to the adjacent bony landmarks.It provides anatomical evidence for lateral ankle ligament reconstruction in treating chronic ankle instability.

AIM: To investigate the effect of Shenmai Injection on the expression of bFGF and PCNA of gastric cancer in mouse. METHODS: RT-PCR was used to measure the expression of bFGF and PCNA in gastric cancer cell cultured with Shenmai Injection in three concentrations.immunohistochemistry used for protein synthesis in mouse gastric cancer model. RESULTS: 140 ?L/mL of Shenmai Injection concentration inhibited 63% of bFGF and PCNA gene expression. CONCLUSION: Shenmai Injection can inhibit the express of bFGF and PCNA.

Objective:To investigate the status of women′s psychological security and quality of life in poverty-stricken areas, and analyze their interrelation and influencing factors.Methods:With the method of convenient sampling, 368 women from poor families who participated in the poverty alleviation vocational training course were investigated with self-designed questionnaire, Psychological Safety Scale (SQ) and World Health Organization Quality of Life-Brief (WHOQOL-BREF).Results:The average total score of women's psychological security in poverty-stricken families was 48.66 ± 12.75, which was in the middle level. Multivariate linear regression analysis showed that family income and religious beliefs had an impact on interpersonal security ( F value was 16.509, P<0.05); ethnicity and religious beliefs had an impact on the certainty of control ( F value was 17.546, p<0.05); religious beliefs had an impact on the total value of security ( F value was 20.911, P<0.05). The overall quality of life and health status scores of females from poor families (13.38±2.91) were significantly lower than the norm ( t value was -3.27, P<0.05). Multiple linear regression analysis showed that personal annual income and religious beliefs had an impact on physiological domain( F value was 2.694, P<0.05); marital status and religious beliefs had an impact on the psychological domain ( F value was 3.305, P<0.05); family annual income and religious beliefs had an impact on the environmental domain ( F value was 2.866, P<0.05) while they had no impact on the social domain. There was a significant positive correlation between psychological security and quality of life. Conclusions:There is a significant positive correlation between the level of psychological security and the quality of life of poor family women. Improving the quality of life of poverty-stricken family women can improve their psychological security.