Objective To observe the effects of bone marrow mesenchymal stem cells(BMMSCs) conditioned-medium (B-CM) on apoptosis in cultured cortical neurons after hypoxia-reoxygenation in vitro in rats.Methods BMMSCs were isolated,cultured and expanded,the culture me-dium was substituted by new medium that contain no serum when cells grew to 90% confluence.After cultured for 24 hours,the medium was collected as B-CM.Cortical neurons of mouse cultured for 8 days were divided into three groups:normal control groups,hypoxic group,B-CM disposed groups.Cell viability were detected by 3-(4,5-dimethlthiazd-2-yl)-2,5-diphenyltetra-zoliu bromidel (MTT) methods and the apoptosis of neurons was examined by flow cytometry and telemicroscopy on hypoxia for 0,6 hours and hypoxia-reoxygenation for 24 hours,respectively.Results Compared with control group,hypoxia for 6 hours could decrease cell viability,hypoxia-reoxygenation for 24 hours increased the cell apoptosis ratio remarkably(P

Objective: To construct a recombinant retroviral vector harboring MafA and to establish a liver epithelial progenitor cell (LEPC) line for stable expression of MafA. Methods: MafA was amplified by PCR and suncloned into pBMN-Z-IRES-Neo vector to obtain pBMH-MafA-Neo vector. After introducing the pBMN-MafA-Neo into Phoenix package cells, the cell culture supernatant was used to infect LEPCs. LEPCs stably expressing MafA gene were screened out. RT-PCR was used to detect the influence of MafA on the phenotype of LEPCs. Results: We successfully constructed pBMH-MafA-Neo vector and obtained LEPCs which stably expressed MafA. Expression of GK and GLUT2 genes in LEPCs-MafA cells was higher than that in the LEPCs. Conclusion: We have successfully obtained LEPCs which can stably express MafA,laying a basis for studying the differentiation of LEPCs into pancreas cells.

Objective: To construct a recombinant retroviral vector expressing pancreatic duodenal homeobox-1 (PDX-1) gene and express the PDX-1 expression cassette in liver progenitor cells (LEPCs), so as to study the conversion of hepatic cells to pancreatic-like cells. Methods: The full length of PDX-1 gene was amplified by PCR and subcloned into pMSCVpuro vector. The pMSCV PDX-1 puro was then introduced into Phoenix package cells and the cell culture supernatant was used for Ping-Pong infection of another packaging cell line PT67 to obtain stable virus-producing cell line. Results: The pMSCV PDX-1 puro vector was successfully constructed and transfected into PT67 cells. The viral supernatants of PT67 cells efficiently infected LEPCs in vitro and the infected LEPCs stably expressed PDX-1 gene. Conclusion: The pMSCV PDX-1 puro vector and PDX-1 expressing LEPCs have been successfully constructed, laying a basis for studying the transdifferentiation of liver progenitor cells to pancreas lineage cells.

Adrenal Gland Neoplasms ; metabolism ; pathology ; surgery ; Adult ; Diagnosis, Differential ; Female ; Hormones, Ectopic ; secretion ; Humans ; Inhibins ; metabolism ; Keratin-8 ; metabolism ; Myelolipoma ; metabolism ; pathology ; surgery ; Ovarian Neoplasms ; metabolism ; pathology ; secretion ; surgery ; Vimentin ; metabolism

Objective To study the relationships between hepatocellular carcinoma (HCC) and the polymorphisms,promoter methylation,and expression of glutathione S-transferases P1 gene (GST) P1 gene.Methods Using methylation-special PCR (MSP),the methylated status of CpG islands of GSTP1 gene in tumor tissues of 53 HCC and its adjacent nontumor tissues were studied.The enzyme activities of GSTP1 were evaluated by ultraviolet colormetry.And using PCR-RFLP,the genetic polymorphisms of the GSTP1 genes of 74 healthy controls and 53 HCC patients were studied.Results The diffe-rences of the frequency of GSTP1 Ile/Ile,Ile/Val and Val/Val genotypes between HCC patients and the normal controls did not reach statistical significance (X~2=0.84,v=2,P=0.656).The frequency of methyla- tion of CpG islands of GSTP1 gene was significantly higher among the HCC tumor tissues when com- pared to the corresponding nontumor tissues (X~2=19.08,P＜0.001),and significantly higher in stageⅢ-Ⅳcases when compared to the stageⅠ-Ⅱcases (X~2=4.84,P=0.028).GSTP1 enzyme activities of cytoplasm in tumor cells were lower significantly than that in the adjacent nontumor tissues (t=2.49, P=0.014),and significantly higher in stageⅠ-Ⅱcases when compared to the stageⅢ-Ⅳcases (t= 2.31,P=0.025).On the other hand,the GSTP1 enzyme activities of cytoplasm in tumor cells with methylated status of GSTP1 gene were significantly lower than that in tumor cells with unmethylation (t=3.50,P=0.001).Conclusion GSTP1 inactivation via CpG island hypermethylation may contrib- ute to the pathogenesis of HCC.

Objective To explore the value of dexamethasone(DEX) for neuronal cell injury and death by observing the effect of DEX on excitatory amino acid(EAA) and monoamine neurotransmitter in cerebral tissue of neonatal rat with hypoxia-ischemia.Methods Hypoxic-ischemic neonatal rat models were established,the levels of EAA and monoamine neurotransmitter in cerebral tissue were analyzed by using capillary electrophoresis and fluorospectrophotometry method.The rats were divided into 4 groups: small dose DEX group pre-treated with DEX(0.5 mg/kg) prior to hypoxia-ischemia,large dose DEX group pre-treated with DEX(10 mg/kg) prior to hypoxia-ischemia,HIE group and shamful operation group.Results The levels of EAA and monoamine neurotransmitter contents in HIE group were significantly higher than those in shamful operation group(P0.05).EAA contents of large dose DEX group greatly decreased compared with HIE group (P

AIM: To investigate the efficacy and safety of low dose mitomycin C (MMC) to prevent haze in trans photorefractive keratectomy (TransPRK) with moderate and high myopia, and to observe the changes of corneal density.METHODS: Sixty-one patients underwent TransPRK with moderate and high myopia.Eyes were divided into research group (0.1g/L MMC for 40s) and control group(0.2g/L MMC for 40s) randomly.There were 21 patients in research group and 40 patients in control group.Cornea epithelial healing time, pain score, visual acuity, manifest refraction, haze and cornea density were analyzed.RESULTS: The epithelial healing time (0.1g/L group: 3.86±1.11d, 0.2g/L group: 4.23±1.27d) and pain score (0.1g/L group: 2.01±0.58, 0.2g/L group: 1.79±0.7) were no significant difference between two groups(P=0.667, P=0.582).It was similar in spherical equivalent at 1mo and 3mo post-operation(0.1g/L group: 0.28±0.25, 0.05±0.23D;0.2g/L group:-0.13±0.17, 0.07±0.22D;P=0.178, P=0.490).The BCVA of control group decreased at 1mo and improved to the same level as pre-operation at 3mo(F=15.847, P<0.001);0.1g/L group showed the same trend, but the changes were no significant difference(F=3.038, P=0.093).There were also no significant difference in Haze between two groups post-operation(z=-0.709, P=0.479;z=-0.478, P=0.633).The change of cornea density was matched with the BCVA (0.1g/L group F=27.399, P=0.001;0.2g/L group F=8.313, P=0.001)and it was similar between two groups.CONCLUSION: The using of low dose MMC to prevent haze in TransPRK with moderate and high myopia is safe and effective.It is therapeutic equivalence to regular dose (0.2g/L).Besides the slit lamp, we can use the corneal density to measure the corneal transparency.

OBJECTIVETo explore the clinical effect of the sacrococcygeal space injection for the treatment of failed back surgery syndrome.

METHODSFrom July 1998 to October 2012,47 patients with failed back surgery syndrome were treated and included 39 males and 8 females with an average age of 61.5 years old ranging from 35 to 89 years old. Among them,41 patients experienced one time of operation, 6 patients with twice of operation. Forty-one patients underwent single,bilateral fenestration or central laminectomy decompression, discectomy. Six patients underwent total laminectomy discectomy and inter body fusion and pedicle screw fixation. All patients were examined by X-ray plain film, CT or MRI before treatment. The anticoagulation was discontinuation before treatment. The needle was put into the sacrococcygeal gap at prone position in the sense of frustration,suction without cerebrospinal fluid and blood,with injection of Mailuoning (Chinese characters: see text) 15 ml. The pain was assessed by VAS before and after treatment. The Oswestry low back pain disability index and survival quality interference degree were evaluated.

RESULTSAt 1 month after treatment,the pain VAS decreased from 59.24 +/- 17.35 before treatment to 19.19 +/- 11.19 after treatment (P < 0.05); The Oswestry low back pain disability index decreased from (41.35 +/- 9.87)% before treatment to (23.17 +/- 17.56)% after treatment (P < 0.05); The survival quality interference degree decreased from 6.5 +/- 2.2 before treatment to 2.6 +/- 1.4 after treatment (P < 0.05).

CONCLUSIONThe sacrococcygeal gap injection for treatment of failed back surgery syndrome has advantages of simple, safe, fewer complications, and low treatment cost.

Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; administration & dosage ; Failed Back Surgery Syndrome ; diagnostic imaging ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Radiography ; Sacrococcygeal Region ; diagnostic imaging

OBJECTIVETo investigate clinical characteristics and pathological changes of tissue surrounding prosthesis after hip and knee arthroplasty.

METHODSA total of 67 patients receiving hip and knee arthroplasty were included in the study and pathological changes of the revision specimens were evaluated by microscopic examination.

RESULTSOf 67 patients, there were 25 males and 42 females (ratio of 0.6) with a mean age of 64 years. There were 42 cases of revision hip prosthesis and 25 cases of knee prosthesis. The primary causes for the revision varied, including 20 cases of infection (29.9%, within 3 months in 9 cases,3 to 24 months in 3 cases and over 24 months in 8 cases), 14 cases of pain (20.9%), 13 cases of loosening of the prosthesis (19:4%), 9 cases of joint stiffness (13.4%), 8 cases of prosthetic dislocation (11.9%), and 3 cases of prosthesis fracture (4.5%). Pathological findings in the tissue surrounding the prostheses included debris reaction, histiocytes, acute inflammatory, chronic non-specific inflammation, pigmented villonodular synovitis (PVNS), "pseudomembranous", calcification, necrosis, sequestrum, etc. These histological changes were frequently admixed.

CONCLUSIONSVarious reasons may lead to hip and knee revision arthroplasty. The main pathological findings include infection, debris granulomas, chronic non-specific inflammatory changes, PVNS. The surgical pathology of the prosthesis provids guidances for clinical treatment and basic research.

Arthroplasty, Replacement, Hip ; Arthroplasty, Replacement, Knee ; Female ; Hip Joint ; pathology ; Humans ; Knee Joint ; pathology ; Knee Prosthesis ; Male ; Middle Aged ; Reoperation ; Synovitis, Pigmented Villonodular ; pathology

Objective To investigate condition of single drug or multiple drug resistance and sensitivities of different combinations of antibiotics in Helicobacter pylori (H.pylori) infected patients of different ages in Jiaxing City,Zhejiang Province.Methods From January 2007 to December 2011,a total of 6280 patients underwent gastroendoscopy examination were enrolled,the average age of them was 50.8 years old.Among them,129 cases were less than 20 years old,1802 cases were between 20 and 40,3016 cases were between 40 and 60,and 1333 cases were over 60.The mucosa of gastric antrum was collected for H.pylori culture.Drug sensitivity of isolated H.pylori strains was tested with metronidazole,amoxicillin and gentamycin,furazolidone,clarithromycin and levofloxacin.x2 test was performed for the comparison of drug resistance among different age groups.Results Among 6280 specimens,H.pylori culture of 2311 cases was positive (positive rate 36.80％).The positive rate of patients aged between 40 and 60 was the highest (38.43％,1159/3016),and that of patients over 60 was the lowest (33.76％,450/1333).The drug resistance rate of metronidazole in patients between 20 and 40 years reached 98.02％ (644/657) and was the highest.The drug resistance rate of clarithromycin in patents over 60 years old was the highest,which was 22.67％(102/450).The drug resistance rates of the levofloxacin,furazolidone,amoxicillin and gentamicin were low of all ages.Mixed resistance of two antibiotic appeared in 586 strains,mixed resistance rate of metronidazole and clarithromycin was the highest (259 strains,15.23 ％).Mixed resistance of three drugs appeared in 49 strains,mixed resistance rate of metronidazole,clarithromycin and levofloxacin was the highest (32 strains,12.88％).A total of 1691 strains were sensitive to the combination of amoxicillin and gentamycin,the sensitive was rate 99.41％.The sensitive rate of the combination of amoxicillin and furazolidone,gentamycin and furazolidone was both 94.24％.Conclusions Metronidazole presented high drug resistance of all ages,and should not be chosen.The combination use of amoxicillin and gentamicin is the ideal antibiotic combination for different ages.The combination of gentamycin and furazolidone is recommended for patients who are allergic to penicillin.