Background Studies showed that some members of matrix metalloproteinases (MMPs) play an important role in the pathogenesis of diabetic retinopathy (DR).However,whether MMPs inhibitor can deter blood retinal barrier (BRB) from damage is below understood.Objective This study was to investigate the effects of GM6001,a MMPs inhibitor,on BRB permeability.Methods Twenty-four adult clean SD rats were randomized to the control group,diabetic group and diabetes+GM6001 group according to randomized number table.Diabetic models were induced by the intraperitoneal injection of streptozotocin in the rats of the diabetic group and the diabetes + GM6001 group,and equal volume of citrate buffer was used in the same way in the rats of the control group.GM6001 10 μ1 (100 μ mol/L) was intravitreously injected in the third and fourteenth day after modeling in the diabetes+ GM6001 group,and equal volume of normal saline solution was injected in the same way in the control group and the diabetic group.The rats were sacrificed and eyeballs were extracted 1 month after injection,and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in rat retinas were detected by reverse transcription PCR (RT-PCR).Evens blue (EB) was infused via the right jugular vein,and paraformaldehyde solution 1％ was then infused via left ventricle at the perfusion pressure 120 mmHg.The eyeballs were extracted 2 minutes later,and the leakage of EB in rat retinas was examined.Results RT-PCR electrophoresis exhibited the response bands of MMP-2 mRNA,and MMP-9 mRNA and GAPDH,with the gene size of 436,536 and 484 bp,respectively.The difference of the MMP2 mRNA and MMP-9 mRNA was statisticaly significant (F =20.336,P =0.000 ; F =8.742,P =0.002) ; and the relative expressions of MMP-2 mRNA and MMP-9 mRNA were significantly higher in the diabetic group and diabetes +GM6001 group than those in the control group (all at P＜0.01),and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in the diabetes+GM6001 group was significantly reduced in comparison with the diabetic group(both P=0.01,P=0.02).The standardized EB content in the retinas of the control group,diabetic group and diabetes+ GM6001 group was (12.60±3.50) ng/mg,(26.52±7.14) ng/mg and (17.55±2.65) ng/mg,showing a significant difference (F=17.032,P＜0.01),and EB content in rat retinas in the diabetic group was higher than that in the control group (P=0.003),and that in the diabetes+GM6001 group was lower in comparison with the diabetic group (P=0.020).Conclusions Intravitreal injection of GM6001 can down-regulate the expression of MMP-2 mRNA and MMP-9 mRNA in diabetic rats and therefore protect BRB.

Lactose was shown to no less competent than Isopropyl-?-D-thiogalactoside (IPTG) in inducing the expression of the ENHANCIN coding gene from Helicoverpa armigera granulosis virus in Eswcherichia coli BL21 (DE3) regulated by a T7 promoter, since the lactose induction could lead to an ENHANCIN band no smaller than the one in IPTG induction on the SDS-PAGE gel. This would decrease the cost of the large-scale ENHANCIN production. The lactose concentration was optimized at 2.2% - 2.5% (w/v) . Different treatments on the lactose sterilization showed that lactose steam- sterilized in 116. 5℃ for 15min could lead to the ENHANCIN production. The convenience and the relatively low cost in its" operation could further decrease the cost of the ENHANCIN production.

Objective Human sperm hyaluronidase in the acrosome is a key enzyme during fertilization.To study and establish a modified substrate film method to improve the diagnotic and treatment of male infertility and investigate the influence of sperm hyaluronidase on male fertility.Methods According to the biochemical feature of sperm hyaluronidase that can dissolve the hyaluronic acid in matrixes,the modified sodium hyaluronate-Gelatin membrane was used as substrate to demonstrate the sperm hyaluronidase activity by incubation and staining.70 human semen samples were selected,categorized and detected for hyaluronidase activity according to the resuh of routine clinical semen examination.The average sperm hyaluronidase activity was statistically anyliezed between the fertile group and infertile group.Results Under general light microscope,the clear and bright digestion circle around the sperm head can be observed against the background of deep purple-blue stained substrate at the positive-reaction area.The amounts of positive reaction and the diameter of the bright circle are positively related to the activity of sperm hyaluronidase.The average hyaluronidase reaction rate and diameter of normal fertile group were 70.84% and 78.17 ?m;What about the infertile group A were 60.02% and 76.92 ?m;The infertile group B were 29.11% and 8.22 ?m; There was statistical difference of HYD activity between the infertile group.B and the fertile group(P

Objective To analysis of incidence and mortality of prostate cancer in urban Guangzhou during 2000-2011 and discuss the trends.Methods The incidence and mortality rates were calculated by analyzing data obtained from Guangzhou Cancer Registration Center.Standardized rates were calculated using the Segi's World Standard Population.According to the characters of morbidity and mortality,ages were divided into three groups of 0-64,65-74 and ≥ 75 years.The average annual percentage change (AAPC) were calculated by Jointpoint Regression Program 4.0.4.Results Altogether 3 029 new cases of prostate cancer and 964 related deaths were registered between 2000 and 2011.The AAPC was 10.01％ (95％ CI:6.00％-14.20％) in crude incidence rate and 8.04％ (95％ CI:6.20％-9.90％) in crude mortality rate,resulting in a total increase of 287.76％ (from 4.82 to 18.69 per 100 000 population) in incidence and 142.90％ (from 2.41 to 5.86 per 100000 population) over the 12 years.There was no significant difference in both standardized incidence and mortality change.The AAPC was 11.51％ (95％ CI:6.00％-17.30％) in 0-64 age incidence rate (resulting in increase of 315.87％ and 1.99 per 100,000 population),4.19％ (95％ CI:0.30％-8.20％) in 65-74 age incidence rate (resulting in increase of 92.72％ and 45.82 per 100 000 population) and 2.77％ (95％CI:-1.00％-6.70％) in ≥75 age incidence rate (resulting in increase of 42.84％ and 63.13 per 100 000 population).The age incidence rate increase was the largest in the ≥ 75 age group,but fastest in the younger group.The age mortality trend was only statistically significant in 0-64 age group,AAPC was 6.25％ (95％ CI:0.30％-12.60％).Conclusions The incidence and mortality rates of Guangzhou increased in a fast rate.The prevention and control of prostate cancer should be strengthened.

Objective To evaluate left ventricular(LV) systolic function by two-dimensional speckle tracking imaging(2D-STI) in severe aortic stenosis(AS) patients.Methods Standard echocardiography and 2D-STI examinations were performed in a total of 54 subjects including 26 consecutive patients with severe AS with LV ejection fraction(LVEF) ≥50％ and 28 controls.2D-STI including systolic longitudinal strain (LS) and strain rate(LSr) were assessed from the apical 4-chamber,3-chamber and 2-chamber views,the circumferential strain(CS) and strain rate(CSr),radial strain(RS) and strain rate(RSr) were measured from the standard short axis views (averaging 6 segments per view).The above parameters of AS groups were compared with those of normals.The repeatability of LVEF,LS and RS was evaluated in 6 AS patients and 6 normal volunteers selected at random from the investigation.Results ① There was no significant difference between AS group and control group in LVEF,LV end-diastolic dimension(LVEDD),LV endsystolic dimension (LVESD) and midwall fractional shortening (mwFS) (P ＞ 0.05).② Significant differences were detected between the two groups.LS,RS,CS,LSr,RSr,and CSr were obviously decreased in AS group(P ＜0.05).③The repeatability of LVEF,LS and RS was good by consecutive measurement of identical and independent observers.Conclusions Despite the presence of normal LVEF,the LV systolic function is impaired in severe AS patients.

Objective To investigate the clinical features and the prognosis after surgical treatment for hilar cholangiocarcinoma (HCC).Methods The surgical therapy and follow-up result were retrospectively analyzed on 98 cases of hilar cholangiocarcinoma admitted into our hospital from January 1995 to January 2005.Differences between groups were evaluated using Chi-square analysis or Student t-test according to the data type.Survival rate was calculated with Kaplan-Meier method,and using the log-rank test.Results Among 98 patients,83 patients underwent surgical treatment (radical resection in 33,palliative resection in 16,and nonresectional internal or external bile duct drainage in 34),15 patients underwent conservative therapy.The 1-,3-,5-year survival rates were 79％,42％,and 17％ in the resection group and 88％,54％,and 24％ in the radical resection group,respectively.The 1-,3-year survival rates were 55％,and 9％ in palliative resection group,respectively,and none of the patient survived for over 5 years.There were significant differences in the survival rate among the radical resection group and the palliative resection group (log-rank test,P ＜ 0.001).Conclusions Radical resection improves the prognosis of hilar cholangiocarcinoma.

OBJECTIVETo summarize the clinical application result of the selective nerve root blocks in limited operation of the lumbar spine.

METHODSFrom January 2008 to October 2012,68 patients with lumbar spinal canal stenosis with multiple levels were underwent the selective nerve root blocks in limited operation of the lumbar spine,including 47 males and 21 females with an average age of 56 years old ranging from 45 to 80. After never roots blocks,64 cases were positive for limited operation of the lumbar spine; the other 4 cases were negative and abort the operation.

RESULTSThe nerve roots block operation smoothly and no complications related to the nerve roots block occurred. There was no neurologic injury complication in this study. Follow-up period ranged from 16 to 45 months postoperatively (means, 32 months). The recovery effect was calculated with Macnab scores, the result was excellent in 44 cases, good in 18 cases, poor in 1.

CONCLUSIONOperative treatment for lumbar spinal canal stenosis with multiple levels is focused on the areas causing symptomate neural compression rather than prophylactic decompression at areas of nonsymptomatic disease. Application of selective nerve root blocks can accurately judge the responsible vertebral body and pain source and improve the curative effect of limited operation of the lumbar spine

Aged ; Aged, 80 and over ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Nerve Block ; methods ; Spinal Nerve Roots ; Spinal Stenosis ; surgery

OBJECTIVETo summarize the clinical application results of the repair soft tissue defect in contralateral leg with a cross-leg soleus muscle flap pedicle transplantation.

METHODSFrom January 2008 to January 2013, 8 patients with soft-tissue defect in lower leg underwent reconstruction with a cross-leg soleus muscle flap pedicle transplantation (without microvascular anastomoses). There were 7 males and 1 female, aged from 20 to 49 years old with an average of 31.8 years. The operative time after injury was from 2 to 8 weeks with the mean of 46 days. The soleus muscle flap was transposed across to the contralateral leg defect area, then immediate to perform the coverage of the muscle flaps by a meshed split-thickness skin graft. The donor site was closed directly.

RESULTSAll the muscle flaps had survived completely. In one case, recipient area edge had a less exudate from drainage hole everyday, the incision spontaneously was healed after 2 week's changing dressing. Follow-up period ranged form 1.5 to 4 years with an average of 2.5 years postoperatively. The tibia and fibula fractures were healed well. A good contour was achieved at the recipient area. According to LEM standard, 2 cases got excellent results, 5 good and 1 fair.

CONCLUSIONSoleus flap pedicle transplantation is very suitable to repair the soft tissue defect of the injuried leg only one main blood vessel, and can reduce the damage of donor area.

Adult ; Female ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Muscle, Skeletal ; Soft Tissue Injuries ; surgery ; Surgical Flaps

OBJECTIVETo investigate the change of expression level of metastasis suppressor gene Kiss-1 in the colorectal cancer cell line SW480 after radiation, and to determine its association with the proliferation and apoptosis of SW480 cells.

METHODSSW480 cells were divided into control group (0 Gy) and study groups (2, 4, 6, 8 Gy). Cells in the study groups were irradiated by 6-MV X-ray radiation for 48 hours. Immunohistochemistry and real-time PCR methods were used to investigate the influence of radiation on Kiss-1 gene expression of SW480. Colony formation assay was used to detect the proliferation of SW480. Flow cytometry-Annexin- V/PI assay was used to observe the change of the apoptosis rate.

RESULTSCompared with the control group, Kiss-1 protein expression increased after radiation of 6, 8 Gy (P<0.05), but no significant changes were observed after radiation of 2, 4 Gy(P>0.05). Kiss-1 gene mRNA level increased after radiation of 2, 4, 6 Gy, while no obvious change was observed for 8 Gy radiation. The apoptosis rates increased for 4, 6, 8 Gy radiation(P<0.05), however, there was no significant difference for 2 Gy radiation (P<0.05).

CONCLUSIONRadiation may increase Kiss-1 gene expression in SW480 cells, which results in decreases proliferation and increases apoptosis in residual surviving cells.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Kisspeptins ; genetics ; metabolism ; radiation effects ; RNA, Messenger ; genetics ; X-Rays

OBJECTIVETo assess the association between genetic polymorphisms of 7 SNPs in PTPN22 and PADI4 genes and susceptibility to rheumatoid arthritis in Yunnan.

METHODSA case-control study was carried out on 192 patients of rheumatoid arthritis and 288 healthy controls. Genotypes of rs33996649 and 1858 loci within PTPN22 gene, and rs11203366 and rs874881 loci within PADI4 gene were determined with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Genotypes of rs1635579, rs2428736 and rs2240340 in PADI4 gene were determined with pyrosequencing.

RESULTSThe frequencies of alleles and genotypes of rs2240340 locus in PADI4 gene showed a significant difference between rheumatoid arthritis and controls in Yunnan population (P U+003C 0.05).

CONCLUSIONOur results suggested that rs2240340 in PADI4 gene is associated with susceptibility to rheumatoid arthritis in Yunnan.

Alleles ; Arthritis, Rheumatoid ; genetics ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Hydrolases ; genetics ; Male ; Polymorphism, Single Nucleotide ; Protein Tyrosine Phosphatase, Non-Receptor Type 22 ; genetics ; Protein-Arginine Deiminases