Objective:To detect the expression level of microRNA-634 (miR-634) in hepatocellular carcinoma (HCC) and its regulatory effect on the common biological behavior of hepatocellular carcinoma cells .Methods: Real-time fluorescence quantitative PCR (RT qPCR) method was used to detect HCC cell lines (HepG2, SMMC7721, BEL7402, bel7404, SNU739), 69 cases of HCC tissues and matching relative quantification of miR-634 paracancerous tissues and analysis of relationship between miR-634 expression and HCC patients gender, age, tumor size, degree of differentiation, child Pugh classification, BCLC staging, portal vein tumor thrombus and liver metastasis , while building a miR-634 eukaryotic expression vector and transfected into hepatocellular carcinoma cell lines, using live cell counting kit-8 CCK-8, flow cytometric annexin V/PI double staining and Transwell experiment to detect the transfection miR-634 on cell proliferation , apoptosis and invasion effects .Results:Compared with the normal human liver cell line L-02 and hepatocellular carcinoma cells miR-634 were decreased ( P<0.05 ) , the expression followed by HepG 2>SNU739>Bel7402>Bel7404>SMMC7721;69 cases of hepatocellular carcinoma ( HCC ) of miR-634 level ( 0.253 ±0.019 ) and lower than that of the matched paracancerous tissues ( P<0.05 ) , and related with the tumor size , degree of differentiation , BCLC stage , portal vein tumor thrombus and liver metastasis ( P<0.05 ) .Over expression in the transfected group 24-96 h after miR-634 level continues to rise , control group and blank vector transfected group differences were statistically significant ( P<0.05 );and the control transfection group and blank group compared to transfection proliferation inhibition rate , apoptosis rate was increased , but wear the number of cell membrane decreased, the difference was statistically significant (P<0.05).Conclusion:miR-634 in hepatocellular carcinoma tissues and cells were low expression and related to clinical pathological parameters , raised its level can inhibit the proliferation and invasion of hepatocellular carcinoma cells and induce apoptosis , for the prevention and treatment of liver cancer has an important reference value .

The involvement of astrocytes and correlation between neuronal injury and astrocyte response were studied. Blockingmiddle cerebral artery and reperfusing o. 5～48 h, H-E staining, immunoccytochemistry single-and double-labeling, dotble label-ing combined with TUNEL and GFAP immunocytochemistry were used to investigate neuronal injury and astrocyte response.The is chemic area peaked at 24 h of reperfusion. The neurons presented irreversible degeneration at 6 h of reperfusion. At24 h,ischemic area in the preoptic area developed into infarcted area; astrocytes exhibited differential morphological features: reactive,malnourished and degenerative changes. At 48 h of reperfusion, the number of astrocytes began to go up. The astrocytes in is-chemic area didn't proliferate within 48 h. By contrast, a few astrocytes underwent apoptosis. In conclusion, these data indicatethat the reaction of astrocytes is closely connected with the extent of neuronal injuries. The reactive astrocytes imply that astro-cytes positively respond to the neuronal injuries, which might play a role in promoting neuronal survival.

Objective To explore the effect of cryopreserved canine kidney cells (MDCK)single -layer on the isolation and proliferation of influenza virus B.Methods Revived P17 MDCK cells were passage for 2 -4 gener-ations,and subsequently preserved in 4℃ refrigerator for 3,6 and 9 days,respectively.Under same experimental con-ditions,the 4℃ refigerater preserved cells were co -incubated with influenza -like illness(ILI)throat swab speci-mens.Cytopathic effect (CPE)was observed,and the proliferation of virus was determined using real -time PCR and the hemagglutinin titers were determined by serological test.Results (1)CPE:The CPE of the MDCK cells pre-served in 4℃ refrigerator for 3 or 6 days had no significant differences compared with that in the control group,while the cell preserved in 4℃ refrigerator for 9 days showed CPE fastly and maintained for a short time.(2)Real -time PCR:the proliferation of influenza virus B in the MDCK cells preserved with 4℃refrigerator for 3 or 6 days (25.86 × 105 -30.25 ×106 ,26.31 ×105 -30.54 ×106 )had on difference compared with that of the control group (24.82 × 105 -29.86 ×106 ),with the proliferation rate of 105 to 106 times,while the proliferation cell with 4℃ cryopreserved for 9 days(19.72 ×104 -28.34 ×105 )the proliferation in cells preserved for 9 days was sharply decreased,with pro-liferation rate of 104 to 105 times.(3)The HA titer:The virus strains with hemagglutination titer above or equal to 116 (P >0.05)isolated with MDCK cells preserved in 4℃ refrigerator 3 or 6 days were not significantly different from that of the control group (10.92 ±0.79).And the cells with 4℃ cryopreserved for 9 days were significantly dicreased (P <0.01).Conclusion No significant effects on the isolation and proliferation of influenza virus B using MDCK cell preserved in 4℃ frigerator for near one week were observed in the present study.

Objective:To report a case of McCune-Albright syndrome (MAS). Methods:Investigated one case's clinical data of McCune-Albright syndrome and reviewed related literatures. Analyzed the cause of disease,clinical manifestation, diagnosis, treatment and prognosis. Results: The diagnostic criteria of MAS are the fibrous dysplasia of bone (FD), in addition to at least a kind of typical hypercrinemia, and (or) special café-au-lait skin spots. A definite diagnosis can be made by discovering the mutation of Gs α-gene in cyst fluid from the ovarian follicle and exceptional bone tissue by gene diagnosis. Conclusion:MAS is a rare disease in clinic and the most of domestic doctors don't know it yet, so it is easy to be misdiagnosed or missed diagnosis. There is no specific treatment for MAS. We must consider it when encounter these patients with sexual precocity as well as with café-au-lait skin spots, and it will be helpful to treat by earlier discovery.

BACKGROUND:Total knee arthroplasty has matured in clinical treatment. LPS-Flex Mobile Bearing System (Zimmer, USA) artificial knee prosthesis is the high-flexion rotating platform type knee prosthesis. The time of its clinical application in China is short, so its advantages have not been reported. OBJECTIVE:To investigate the preliminary clinical outcome of the total knee arthroplasty with LPS-Flex Mobile Bearing system artificial knee prosthesis (Zimmer, USA), and to assess the biocompatibility of artificial prosthesis and host using range of motion of knee and function after replacement. METHODS:We retrospectively analyzed 37 patients (42 knees) undergoing total knee arthroplasty using Zimmer LPS-Flex Mobile Bearing prostheses (high-flexion rotating platform type knee prosthesis) in The First Hospital Affiliated to Soochow University from February 2012 to March 2014, including 9 males (10 knees) and 28 females (32 knees), aged 47-78 years, averagely 63.7 years. Bone cement fixation was used, and the patel a was not treated with replacement. Postoperative complications were observed. Ranges of motion of knee joint preoperatively and postoperatively were compared. The recovery of knee joint function was evaluated using Hospital for Special Surgery Knee Score. RESULTS AND CONCLUSION:A total of 34 cases (38 knees) were fol owed up for 6-28 months. Range of motion of knee joint improved from 88.5° before operation to 124.2° after operation on average. Hospital for Special Surgery Knee Score improved from 52.5 before replacement to 91.1 after replacement, showing significant differences (P<0.01). Therapeutic effects were assessed according to Hospital for Special Surgery Knee Score:excel ent in 20 cases, good in 16 cases, average in 2 cases, with an excel ent and good rate of 95%. The incidence of various complications was low. These data suggested that short-period clinical outcomes of high-flexion rotating platform type knee prosthesis replacement are satisfactory. This prosthesis has advantages in its design, which is more close to the physical structure of knee joint, but its long-period outcomes deserve further investigations.

Objective To study the composition of variant and point mutations of Norovirus GⅡ.4 in Guiyang regions.Methods From June to November 2010,cases information and fecal specimens were collected from guard-hospitals in Guiyang regions,who had caught the acute-gastroenteritis.Noroviruses in specimens were detected by a real-time reverse transcription polymerase chain reaction(real-time RT-PCR),and then partial genotyped norovirus-positive clinical samples (in random) were cloned and sequenced in VP1 gene code.Furthermore,the gene sequences were compared with the published variants at home and abroad of norovirus(GⅡ.4),including the phylogenetic analyses of genomes and variation of amino acids within individual sites.Results Those 267 specimens were GⅡ-norovirus-positive(62.68％) in 426 clinical samples.There were nine GⅡ.4-norovirus-positive VP1 gene-sequences available,and two subtype-norovirus variants (GⅡ.4 2008a and G Ⅱ.4 2008b variant) were epidemic in 2010,Guizhou province.The homology between and in subgroups were 95.90％-96.72％ and 99.45％-100％.Two amino acids within individual sites were apt to mutate.Conclusion Norovirus GⅡ genotype were predominant in summer and fall acute gastroenteritis in 2010 for Guiyang regions,and the variants were diversity.

Objective To evaluate the inhibitory effects of shRNAs targeting genes encoding viral capsomeres VP1-VP4 on enterovirus 71 (EV71) replication when used alone or in combination.Methods Short hairpin RNAs (shRNAs) targeting genes encoding VP1-VP4 protein of EV71 were designed and then respectively inserted into lentiviral vector pLKD-CMV-GFP-U6 to construct the recombinant plasmids.The expression plasmids together with psPAX2 and pMD2.G were transfected into 293T cells to induce the expression of recombinant lentiviruses,which were collected on the third day after transfection.The titers of recombined lentiviruses were determined by real-time PCR.The effects of shRNAs used alone or in combination on the expression of EV71 at mRNA and protein levels were respectively detected by real-time PCR and Western blot.Results The inhibitory effects of shRNAs on EV71 replication showed no significant differences among various strains (isolated from fatal cases,severe cases,mild cases and FY0805) (P＞0.05).Their inhibition rates were 51.6％ (sh-VP1-1),85.1％ (sh-VP1-2),76.4％ (sh-VP1-3),57.5％ (sh-VP2-1),81.4％ (sh-VP2-2),79.5％ (sh-VP2-3),68.9％ (sh-VP3) and 56.7％ (sh-VP4) respectively,and they were in a dosage dependent manner.sh-VP1-2 in combination with sh-VP2-2 showed the highest inhibition rate reaching up to 96.6％.Moreover,shRNAs used in combination showed better effects than any one used alone even at double dosage.Conclusion All shRNAs targeting viral capsid VP1-VP4 genes showed inhibitory effects on EV71 replication with inhibition rates over 50％ and the effects could be strengthened when using shRNAs in combination.

BACKGROUND:Open fracture of lower limb with severe soft tissue and bone defects also accompanies anterior tibial soft tissue defects and exposure of sclerotin and steel plate, which can be crucial y treated with strong fixation and use of skin flap to block the wound.
OBJECTIVE:To explore the clinical efficacy of a large area of soft tissue defects in the anterior tibia using vacuum sealing drainage combined with groin free flap.
METHODS:A total of 24 patients with a large area of soft tissue defects in the anterior tibia were included in this study and then divided into two groups, with 12 cases in each group. In vacuum sealing drainage group, the scope of soft tissue defects was ranged from 10 cm×15 cm to 15 cm×20 cm. After the debridement, the fracture was fixed with external fixation scaffold and the wound was covered with the vacuum sealing drainage dressing. The blood clot was rinsed with normal saline via T-tube, and 7-10 days later the vacuum sealing drainage was given. According to the growth of granulation tissue, the wound was secondarily sutured, fol owed by groin free skin flap of superficial iliac circumflex artery with medial knee arteriovenous anastomosis transplantation. In the non-vacuum sealing drainage group, the wound size was ranged from 10 cm×5 cm to 30 cm×20 cm, the period from injury to admission was 1-24 hours. They were given conventional debridement and secondary fixation or skin flap transplantation.
RESULTS AND CONCLUSION:The length of preoperative hospital stay and the skin flap are in vacuum sealing drainage group were significantly better than those in non-vacuum sealing drainage group (P<0.05). There was no significant difference in the length of postoperative stay and total length of hospital stay between the two groups (P>0.05). The wound infection rate was 0 in vacuum sealing drainage group and 75%in non-vacuum sealing drainage group at 8-14 days after treatment. The wound and donor area incision were healed at I stage, the skin grafts survived. Al the involved patients in two groups were fol owed up, for 6-36 months. During the fol ow-up process, the fracture healing time in non-vacuum sealing drainage group was significantly longer than that in vacuum sealing drainage group. The skin flap in two groups was similar to surrounding skin in color and texture, the flap exhibited no vessels, no ulceration, and no clumsy. The vacuum sealing drainage combined with groin free flap can timely control a large area of soft tissue defects post-trauma, improve wound blood supply, shorten preoperative preparation time, early close the wound, significantly promote the healing of wound and fracture. The skin flap is soft, flexible, wel-looking, and active functional, it significantly shortens the course of treatment and maximizes the recovery of limb function.

Eight patients with suspected cases of C .jejuni were etiologically diagnosed and analyzed in this study to pro-vide scientific basis for the confirmation of the cases of human campylobacteriosis in Guizhou Province ,China .Blood or feces of 8 suspected patients were employed to isolate bacteria strains .Conventional and multi-PCR techniques were applied to identify suspicious bacteria strains .The C .jejuni strains were analyzed by using Pulsed Field Gel Electrophoresis (PFGE) .Suspicious strains of C .jejuni were isolated from all the 8 suspected patients of campylobacteriosis and anticipated genes fragment were detected with multi-PCR .With the digestion of restriction enzyme SmaI ,the 8 C .jejuni strains were divided into 7 PFGE pat-terns with 7-10 DNA bands .Cluster analysis showed that the gross similarity of 8 strains of C . jejuni was more than 50% . The similarity of PFGE patterns between strain GZ201004 and GZ201005 from diarrhea patients was as high as 100% ,while the similarity of strain GZ201201 and GZ201007 was 66 .7% .Moreover ,C . jejuni were detected from all the suspected pa-tients of campylobacteriosis .PFGE results indicated that strains GZ201004 and GZ201005 were from the same source ,while all the 8 isolates showed PFGE polymorphism .

Objective:To study the effect of microRNA (miR)-330-3p on hepatic ischemia-reperfusion injury (IRI) in mice, meanwhile, and to determine potential molecular mechanism.Methods:Eighty male C57BL/6 mice, aged 7-8 weeks, 23-25 g, specific pathogen free, were randomly divided into 8 groups (10 mice in each group) using random number table: reperfusion 2 h group, 6 h group, 12 h group, 24 h group, sham group, miR-330-3p agomir group (preoperative injection of agonist), miR-330-3p antagomir group (preoperative injection of inhibitor) and miRNA-NC group. Except for the sham group, the hepatic IRI model were established in mice. Polymerase chain reaction (PCR), Western blot and immunohistochemistry were used to detect the expression of miR-330-3p and phosphoglycerate mutase family member 5 (PGAM5), cleave caspase-1 and GSDMD. Luciferase reporter assay was performed to investigate whether miR-330-3p directly targets PGAM5. At the same time, AML12 cells were also treated with miR-330-3p mimics/inhibitor or PGAM5 siRNA, then the expression of PGAM5, NLRP3, cleave caspase-1 and GSDMD were detected by Western blot analysis.Results:Level of miR-330-3p gradually decreased after reperfusion, however, mRNA level of PGAM5 was increased thereafter ( P<0.05) as compared with the sham group. Serum level of AST and ALT were decreased in miR-330-3p agomir group while that of were increased in miR-330-3p antagomir group as a function of time following reperfusion, and the differences were statistically significant (all P<0.05). Cleave caspase-1 expression was decreased in miR-330-3p agomir group but was increased in miR-330-5p antagomir group ( P<0.05). Luciferase reporter assay was performed to determine PGAM5 was a target gene of miR-330-3p. SiRNA-mediated knockdown of PGAM5 decreased level of GAM5 (0.24±0.09), NLRP3(0.12±0.07), cleave caspase-1 (0.15±0.07) and GSDMD (1.08±0.08) as compared with the siRNA-NC group (1.17±0.14), (0.36±0.09), (0.68±0.09), (1.36±0.08), and the differences were statistically significant (all P<0.05). Conclusion:MiR-330-3p can alleviate hepatic IRI in mice, which may be related to inhibition of PGAM5-induced pyroptosis.