Objective To explore the effect of different antifreeze or antioxidant at different cryopreservation conditions on the activity and the morphology of adipose cells.Methods After purification,fat granules were divided into 5 groups; the same volume of antifreeze or antioxidants were added in each group,stored at-20℃ (refrigerator),-80℃ (ultra-low-temperature refrigerator) and-196 ℃ (liquid nitrogen) ; the amount of glucose transfer was determined and the morphological changes were observed for each group after two weeks,two months,and three months.Results Fat activity of the group adding both dimethyl sulfoxide (DMSO) and trehalose was significantly higher than that in other groups at the same temperature and the same time point.The decreased amount of glucose transfer of 80℃ group and 196℃ group was significantly lower than that of 20℃ group,and that in groups of each 196℃ were greater than that in groups of each 80℃.Conclusions Antifreeze can obviously keep activity of fat cells.Preservative effect of DMSO is better than trehalose,and that of both combination is much better.Liquid nitrogen storage of adipose tissue for three months and fat granules stored at 80℃ for two months are still suitable for clinical use.

砄bjective:To investigate the expression of integrin ? 1 during mandibular fracture healing.Methods:Rabbit mandibular fracture model was used,the fractured bone tissues were obtained and paraffin slices were made on the day 1,3,5,7,14,30,60 and 90 after surgery respectively.Non fractured mandible was used as the control.The expression of integrin ? 1 in bone tissue, especially in osteoblasts and osteoclasts, was studied with LsAB immunohistochemical method.Results:Integrin ? 1 was widely espressed in the bone tissure close to fracture and surrounding soft tissue.In osteoblasts increased expression of integrin ? 1 was observed on day 7,maximum on day 14～30 and normal on day 90.While in osteoclasts,it was increased and kept at maximum 14～90 days aftere fracture.Conclusion:During mandibular fracture healing,the expression of integrin ? 1 in bone,especially in osteoblasts and osteoclasts,was increased.Integrin ? 1 has a critical influence on bone fracture healing.

Objective To study the changes of cytokines in children with hand-foot and mouth disease.Methods Sixty-two cases of hand-foot-mouth disease admitted to our hospital from March 2011 to December 2011 were included in this study.Thirty-seven cases were male and 25 cases were female,the youngest one was four month old,and the oldest one was eleven month and 4 years old,the average age was (1.56 ± 0.98) years old.Sixty-two children with hand-foot-mouth disease were devided into ordinary group (n =20),severe group (n =20),critically ill group (n =22).Twenty healthy children served as control group.The plasma levels of pro-inflammatory cytokines [tumor necrosis factor(TNF)-α,interleukin(IL)-1β、IL-6] and anti-inflammatory cytokines (IL-1 0、IL-13、IL-4) were detected.Results The pro-inflammatory and anti-inflammatory cytokines of patients with hand-foot and mouth disease were increased compared with control group.The levels of TNF-α in severe and critically ill groups were (201.45 ± 133.42) ng/L,(220.37 ± 117.54)ng/L respectively,which significantly increased compared with ordinary group [(118.12 ±11205) ng/L] (P ＜0.05).The levels of IL-1β and IL-6 in critically ill cases were (651.77 ±538.45) ng/L and (88.02 ±50.76) ng/L respectively,which significantly increased compared with severe cases [(551.75 ± 252.75) ng/L,(59.25 ± 28.80) ng/L] and ordinary cases [(270.18 ± 249.79) ng/L,(55.76 ± 28.68) ng/L] (P ＜ 0.05).The level of IL-10 was (137.53 ± 72.07) ng/L in ordinary cases,which increased significantly compared with the severe and critically ill cases[(57.07 ±55.02) ng/L,(73.37 ±56.01) ng/L] (P ＜0.05).The levels of IL-13 were (507.53 ±210.86) ng/L and (489.67 ± 168.99) ng/L in severe and critically ill cases respectively,which significantly increased compared with ordinary cases [(387.16± 157.43) ng/L] (P ＜0.05).The levels of IL-4 showed no significant differences among the ordinary,severe and critically ill groups(P ＞ 0.05).Conclusion The pro-and anti-inflammatory cytokines were involved in inflammatory response simultaneously in patients with hand-foot and mouth disease.The increases of TNF-α,IL-1β and IL-6 are consistent with severity of the disease,while factor IL-10,IL-13 are decreased as the severity of the disease.The increase of the pro-inflammatory cytokines and decrease of the anti-inflammatory cytokines are the important sign of the exacerbation.

Medical Jurisprudence is an intersection discipline of medicine and Jurisprudence.Luzhou Medical College is the second one that has established jurisprudence discipline ( medical jurisprudence ) in China,and also the first one in the West of China,although college did not get into the subject field in last century.After more than 10 years of exploration and construction,our college has had an innovation way of medical jurisprudence teaching,scientific research,academic exchanges and social service.By integrating the resources in and out of college,gathering excellent talents,setting up reasonable academic team,promoting sustainable development of the discipline,establishing Sichuan Province Medical Jurisprudence Research Center,sponsoring an academic journal Medicine & Jurisprudence,building academic platforms,condensing subject direction,conducting scientific research,serving local economy and society,innovating models,optimizing plans,cultivating medical jurisprudence practice talents,our discipline construction is in the front rank in China,and it has obvious features and advantages,and also has some great influence on the field of medical jurisprudence.

Duck circovirus(DuCV),a potential immunosuppressive virus,was investigated in Southern China from March 2006 to December 2009 by using a polymerase chain reaction(PCR)based method. In this study,a total of 138 sick or dead duck samples from 18 different farms were examined with an average DuCV infection rate of～35%. It was found that ducks between the ages of 40～60 days were more susceptible to DuCV. There was no evidence showing that the DuCV virus was capable of vertical transmission. Farms with positive PCR results exhibited no regularly apparent clinical abnormalities such as feathering disorders,growth retardation or lower-than-average weight. The complete genomes of 9. strains from Fujian Province and 1 from Zhejiang Province were sequenced and analyzed. The 10 DuCV genomes,compared with others genomes downloaded from GenBank,ranged in size from 1988 to 1996 base pairs,with sequence identities ranging from 83.2% to 99.8%. Phylogenetic analysis based on genome sequences demonstrated that DuCVs can be divided into two distinct genetic genotypes,Group I(the Euro-USA lineage)and Group II(the Taiwan lineage),with approximately 10.0% genetic difference between the two types. Molecular epidemiological data suggest there is no obvious difference among DuCV strains isolated from different geographic locations or different species,including Duck,Muscovy duck,Mule duck,Cheery duck,Mulard duck and Pekin duck.

BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

Three SNaPshot multiplex assays were developed to test 23 coding region single nucleotide polymorphisms (SNPs) and one control region SNP outside hypervariable regions (HVR)I and II, which was aimed at increasing the discrimination power of the mitochondrial DNA (mtDNA) typing in forensic casework, and confirming haplogroup assignments of mtDNA profiles in both human population studies and medical research. The selected SNPs targeted the East Asian phylogeny. These multiplex assays were validated by comparing with the sequencing analysis of samples chosen randomly. The mtDNA variations of 100 unrelated individuals from the Wuhan population in China were examined and classified into 31 haplotypes, and the haplotype diversity was estimated to be 0.952. The multiplex SNaPshot method is rapid and robust, and suitable for large-scale screening studies of mtDNA variability.

OBJECTIVETo investigate the expression change of integrin alpha 5 during mandibular fracture healing.

METHODSUsing rabbit mandibular fracture model, the fractured bone tissues were obtained and paraffin slices were made on the days of 1, 3, 5, 7, 14, 30, 60 and 90 after surgery, respectively. Non-fractured mandibles were used as normal control. LsAB immunohistochemical method was used to detect the expression of integrin alpha 5 in bone tissue, especially on osteoblasts and osteoclasts.

RESULTSIntegrin alpha 5 was widely expressed in fractured bone and surrounding soft tissues. The expression of integrin alpha 5 increased 7 days after fracture, peaked in 14 to 30 days and returned to nearly normal in 60-90 days.

CONCLUSIONDuring mandibular fracture healing, the expression of integrin alpha 5 in bone tissue increased clearly. It is estimated that integrin alpha 5 players an important role in fracture healing.

Animals ; Female ; Fracture Healing ; Immunohistochemistry ; In Situ Hybridization ; Integrin beta1 ; biosynthesis ; genetics ; Male ; Mandibular Fractures ; metabolism ; Osteoblasts ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits

Objective As important transcription factors, SOX2, TWIST and SNAIL are closely related to breast cancer me?tastasis. The aim of this study was to explore the expressions of SOX2, TWIST and SNAIL in breast cancer and the relationship between the three factors. Methods We collected 74 specimens of breast cancer and another 74 from paired normal tissue from January 2012 to January 2013. Using immunohistochemistry, we determined and compared the expression of SOX2 in the breast cancer and normal tissues. We also detected the expressions of TWIST and SNAIL proteins and analyzed the relationship of SOX2, TWIST and SNAIL ex?pressions with the clinicopathological characteristics of breast cancer as well as the correlation between the three proteins. Results The expression of SOX2 was significantly higher in the breast cancer than in the normal tissues (25.7% vs 8.1%, P=0.004) and corre?lated with the grade, lymph node metastasis, and TNM stage of breast cancer ( P<0.05) , while the expressions of TWIST and SNAIL were significantly correlated with lymph node metastasis and clinical TNM stages of the malignancy ( P<0.05) . The protein expression of TWIST was correlated positively with those of SOX2 ( Cc=0. 325, P<0.005) and SNAIL (Cc=0.308, P<0.008), and so was that of SNAIL with that of SOX2 (Cc=0.275, P<0.018). Conclusion The SOX2 protein is highly expressed in breast cancer. The expres?sions of SOX2, TWIST and SNAIL are closely related to lymph node metastasis of the malignancy and the three factors are interacted coor?dinatively in the metastasis of breast cancer.

Objective:To establish a method for determining three residual organic solvents in nimodipine liposomes. Methods:The samples were injected into a DB-624 capillary column (30 m × 0. 32 nm,1. 8 μm) by a headspace sampler and analyzed with an FID detector, the carrier gas was nitrogen, the injector temperature was 250℃, and the detector temperature was 250℃. The column temperature was programmed raised. Results:Three residual solvents, namely ethanol, acetone and acetic ether were completely sepa-rated. There was a good linearity within the experimental concentration range. The average recovery was 98. 9%,98. 5% and 99. 4%(RSD=0. 32%,1. 12%,0. 76%,n=9), respectively. The detection limits was 0. 20, 0. 18 and 0. 22μg·ml-1, respectively . Con-clusion:The method is rapid, sensitive and accurate. It can be used in the determination of residual organic solvents in nimodipine li-posomes.