Objective:To study the application of petrifilm colony count plate in the microbial detection for cosmetics. Methods:Three common microbials in 14 batches of cosmetics were respectively detected by the method described in hygienic standard for cos-metics and petrifilm colony count plate, and the results were compared. Results:The results shown by the two methods had no statisti-cally significant difference (P>0. 05). Conclusion: Petrifilm colony count plate with light weight, fast and easy operation exhibits significance in the microbial detection for cosmetics.

The upper limbs of 23 adult cadavers were dissected and the internal structures of the median nerve were studied for microsurgery of the nerve.Under operative micros- cope(24X),the median nerves of 20 cadavers treated with 10% acetic acid were dissec- ted.The median nerves of the remaining three were cross-sectioned,inch by inch,and stained with Sudan Ⅲ for microscopic observation.Microscopic ptotographs were taken to visualize the intra-neural topography of motor bundles or bundle groups. 1.The morphology,topographic relationships,size and number of bundles of the median nerves were observed under microscope and were recorded. 2.At the level of the arm and forearm,the nerve trunk and its internal bundles appear to twist to a certain degree in the way of external rotation from the distal to proximal. 3.The motor bundles of the thenar muscles emerge at the level about 40.1mm below the styloid process of radius.They enter the nerve trunk anteriorly on the radial side,and run upward at the Rower part of the forearm,and continue postero-laterally and appear as a admixture of sensory and motor nerves.At the mid-portion of the fore- arm,they are situated posteriorly,while at the level of antecubital fossa and above, they occupy the central position and finally most of the bundles join the medial head of the median nerve. 4.The motor bundles that supply the flexor muscles of the forearm are located anteriorly and posteriorly in separate portions after entering the nerve trunk.At the level of the arm,these two portions of median nerve are mixed bundles but motor nerve fibers predominate. 5.The fundamental method of funicular suture were discussed in relation to the intra-neural topography of the median nerve.

Objective To investigate the effects of chitosan/pCDNA3.1 (+) CrmA on matrix metalloproteinase (MMP)-1 and tissue inhibitor of metalloproteinase (TIMP)-1 expression of chondrocytes induced by interleukin-1β (IL-1β) in mRNA and protein levels.Methods Rabbit chondrocytes were isolated and cultured.Chondrocytes were treated with PBS,10 μg/ml chitosan (CS) and chitosan/ pCDNA3.1 (+)CrmA respectively for 6 hours.Then 10 ng/ml IL-1β was added into the culture medium.After 48 hours,real time polymerase chain reaction (real time PCR) and Western blot assay were used to examine the changes of MMP1 and TIMP-1 in mRNA and protein levels.Results In CS/pCDNA3.1 (+)CrmA treated group (0.44±0.04),the messenger RNA expression of MMP-1 in chondrocytes was significantly suppressed compared with corresponding samples of PBS treated group (1.00±0.05) and CS treated group (0.76±0.07),There was significant difference between three groups (F=106.93,P＜0.01).The MMP-1 protein expression of chondrocytes in CS/pCDNA3.1 (+)CrmA treated group (0.28±0.03) was lower than that of PBS treated group (0.73±0.06) and CS treated group (0.46±0.05)(F=59.66,P＜0.01).No significant difference of TIMP-1 expression among the three groups was observed in mRNA and protein levels.Conclusion CS/pCDNA3.1(+) CrmA can significantly inhibit the mRNA and protein expressions of MMP-1 of chondrocytes induced by IL-1β,which leads to up-regulation the ratio of TIMPs to MMPs of IL-1β induced chondrocytes.It may be a part of the mechanisms of the therapeutic effects of CS/pCDNA3.1 (+)CrmA on osteoarthritis.

Objective:To establish a method for the microbial limit test of Jingzhi Guanxin tablets. Methods: According to the methods in the 2010 edition and 2015 edition of Chinese Pharmacopoeia, the test was performed respectively. Results:Jingzhi Guanxin tablets showed obvious inhibitory effect on Bacillus subtilis. The bacteria count could be carried out by the culture medium diluting methods in the 2010 edition. The total amount of aerobe could be detected by the membrane filtration method in the 2015 edition. The total combined molds and yeasts count could be performed by the plate count method and the specified microorganism could be tested with the routine method. Conclusion:The above methods can be used for the microbial limit test of Jingzhi Guanxin tablets.

AIM: To investigate the expression of MAT1 protein in pancreatic cancers and the relationship between MAT1 and clinicopathological features of pancreatic cancers. METHODS: 94 surgical specimens, including 70 pancreatic cancers, 10 pancreatic benign tumors, 14 chronic pancreatitis and 10 autopsy normal pancreas tissues, were analyzed immunohistochemically, and then MAT1 expression and clinicopathological features were compared. RESULTS: MAT1 was expressed mainly in the cancer cells,and also in the fibroblasts, where it was localized within the cytoplasm and nuclear envelope. MAT1 expression was found in 75.7% (53/70) of the cancers, but not detected or weakly expressed in control tissues. There was a significant difference in expression of MAT1 among the above four tissues (P

Objective To investigate the effect of tail vein injection of exogenous bone marrow mesenchymal stem cells (BMSCs) on proliferative,osteogenic and adipogenic potential of BMSCs in ovariectomized osteoporotic rats.Methods The animal model of osteoporosis (OP) was established 3 months after ovariectomy (bilateral ovarian resection)in healthy 10-week-old female SD rats with the same batch and weight.The healthy 8-week-old SD GFP rats were selected in the same way as the BMSCs source.The experiment was divided into BMSCs group and control group.The BMSCs (1×106) marked with GFP were injected into BMSCs group rats by tail vein injection.One week after injection,BMSCs were isolated,cultured and purified in vitro to the 3rd,4th passage in all groups.The MTT method was used to detect the growth curves of BMSCs.The oil red O dyeing and alizarin red staining (ARS) were used to compare the adipogenic and osteogenic potential between the two kinds of BMSCs,respectively.The RT-PCR was used to detect the expression of osteogenesis-related genes (Runx2,osteocalcin and osteopontin) and adipogenesis-related genes (PPARγ and LPL) in BMSCs.Results Compared with the control group,the growth curve of BMSCs in BMSCs group rose significantly; the calcified nodule increased significantly after osteogenic induction; the lipid droplet decreased significantly after adipogenic induction.RT-PCR results showed that the Runx2,osteocalcin and osteopontin mRNA expression increased,while the peroxisome PPARγ and LPL mRNA expression decreased in BMSCs group.Conclution By tail vein injection of exogenous BMSCs,the proliferative and osteogenic potential of BMSCs in ovariectomized osteoporotic rats is enhanced,while the adipogenic potential is reduced,which provides a possibility of stem cell therapy for osteoporosis.

Objective:To establish a Taqman fluorescence quantitative PCR for the detection of Salmonella in drugs. Methods:Based on Salmonella specific gene, a pair of primers and a probe were designed, and DNA of Salmonella was extracted for the detec-tion. Results:The method was much specific, and the detection limit was 160 cfu/ml. The detection rate was 100% for the artificially contaminated samples. Conclusion:Fluorescence quantitative PCR can be applied in the rapid detection of Salmonella in drugs.

Objective To investigate the effects of chitosan(CS)/pcDNA3.1(+) CrmA on the expression of interleukin-13 (IL-1β3) converting enzyme (ICE) and IL-1β in chondrocytes.Methods Rabbit chondrocytes were isolated and cultured.Chondrocytes were treated with PBS,10 μg/ml CS/pcDNA3.1(+) and CS/pcDNA3.1 (+) CrmA respectively for 6 hours.Then 10 ng/ml IL-1β was added into the culture medium.After 48 hours,the messenger RNA and protein expression of ICE and IL-1β in chondrocytes were detected by using real time polymerase chain reaction and western blotting.Results In CS/pcDNA3.1 (+) CrmA treated group (0.52 ±0.09),the mRNA expression of ICE inchondrocytes was significantly inhibited compared withcorresponding samples of CS/pcDNA3.1 (+) group (0.84±0.11,t=4.42,P＜0.01) and PBS group (1.00±0.10,t=6.58,P＜0.01).ICE protein expression in CS/pcDNA3.1 (+) CrmA treated group (0.20±0.03) was markedly lower than CS/pcDNA3.1 (+) (0.37±0.05,t=4.85,P＜0.01) and PBS treated group (0.44±0.07,t=6.68,P＜0.01).There was no significant difference of ICE mRNA and protein expressions in chondrocytes between CS/pcDNA3.1(+) group and PBS group.Significant difference of IL-1β mRNA expression was found in the three groups.IL-1 β mRNA expression level was significantly lower in CS/pcDNA3.1(+) CrmA treated group (0.55± 0.08) than CS/pcDNA3.1(+) (0.69±0.06,t=3.50,P＜0.01) and PBS group (0.99±0.04,t=11.12,P＜0.01) of chondrocytes.IL-1β protein expression level of chondrocytesin CS/pcDNA3.1(+) CrmA group (0.230±0.020) was significantly lower than CS/pcDNA3.1 (+) (0.450±0.060,t=5.07,P＜0.01)and PBS groups (0.610±0.090,t=8.70,P＜0.01) of cells.Significant difference of IL-1β mRNA and protein expression between CS/pcDNA3.1 (+) and PBS group was also observed (t=7.61,P＜0.01;t=3.63,P＜0.01).Conclusion CrmA mediated by chitosan can significantly suppress the mRNA and protein expression of ICE,thus down regulat the expression of IL-1β,which may be one of the mechanisms of CS/pcDNA3.1 (+) CrmA in the treatment of osteoarthritic cartilage degeneration.

Objective To prepare and evaluate 99Tcm radiolabeled Cetuximab (C225) for imaging of EGFR specific binding.Methods Cetuximab antibody was reduced by 2-iminothiophene (2-IT).The radiolabeling of IT-Cetuximab with 99Tcm(99Tcm-IT-Cetuximab) was analyzed by HPLC,and was tested for in vitro stability and molecular integrity.The human lung cancer line (A549)-bearing nude mouse model was prepared for biodistribution and tumor targeted study.Tumor uptake was also measured by in vivo γ imaging.Results The labeling efficiency was 93.15％.The radiochemical purity was 96.46％ after purification.The in vitro stability was good in 5％ HSA,in which the radiochemical purity maintained above 80％ at 4 ℃ for 24 h.99Tcm-IT-Cetuximab showed good specific binding to tumor with peak uptake of (3.417±0.769) ％ID/g after 4 h.The T/NT ratio of blood increased to 1.454±0.174 at 24 h.γ imaging of A549-bearing nude mice xenografts also showed high T/NT ratio.Conclusions 99Tcm-IT-Cetuximab molecular probe could be prepared with high radiolabeling yield and radiochemical purity.It has excellent in vitro and in vivo stability,and shows specific uptake in A549 tumor.

AIM: To elucidate the pattern of 5-fluorocytosine(5-FC) induced apoptosis and its role in gene therapy for human pancreatic cancer. METHODS: The human pancreatic cancer SW1990 cells(CEA-producing) were infected with recombinant adenoviruses(Adex1CEA-prCD or Adex1CEA-prZ).Cytosine deaminase(CD) expression was examind by western blot. Apoptosis induced by 5-FC in human pancreatic cancer SW1990 cells genetically modified to express cytosine deaminase was investigated by applying electron microscopy, DNA electrophoresis and flow cytometry analysis techniques. RESULTS: The SW1990 cells infected with Adex1 CEA-prCD were treated with 5-FC at 100 ?mol?L -1 for 48 h, cell apoptosis occurred. Typical apoptosis morphological feature appeared and DNA ladder could be demonstrated on DNA electrophoresis. Apoptosis peak was also showed by flow cytometry. Apoptotic cells accounted for 34.6% of the cell population. Cells in G 1, S and G 2/M phase of cell cycle were 64%, 11% and 7%, respectively. CONCLUSION: The apoptosis induced by 5-FC may be a primary mechanism in CD gene therapy for pancreatic cancer.