BACKGROUND: The commonly used culture methods for primary intervertebral disc cells are type Ⅱcol agenase alone digestion method, and type Ⅱ col agenase combined trypin digestion method. However, the acquired cells are few. OBJECTIVE: To acquire nucleus pulposus cells from human degenerative intervertebral disc using a systemic and simplified method. METHODS: Nucleus pulposus cells were isolated and cultured. The morphology of nucleus pulposus cells was observed under inverted phase contrast microscope every day. Primary and subcultured cellsuspension was applied for the determination of cel viability using trypan blue staining. The cel growth was detected with MTT assay. The cel morphology was observed under laser confocal microscopy. RESULTS AND CONCLUSION: Nucleus pulposus cells were successful y isolated from from human degenerative intervertebral disc using the improved method, and cells were subcultured to passage 3. Primary cells were fusiform shaped, while cells at passage 1 and 2 were triangle or polygonal, which were similar to fibroblasts. When cells almost reached the confluence, the cells showed slabstone-like appearance. Trypan blue staining showed that, the primary cel viability was 99%, and passage 3 cells had 93%-95% viability. The proliferation of nucleus pulposus cells gradual y decreased as the generation increased. Compared with passage 1 cells, passage 2 and 3 cells at logarithmic phase trended to be smoother. The cel morphology observed under laser confocal microscopy was similar to the results under phase contrast microscope. The improved simple method can successful y acquire a variety of cells from human degenerative intervertebral disc, and these cells show fine biological properties.

Objective To compare the values of a screening methods by OC-Hemodia and chemical fecal occult blood test (COB) for colorectal cancer in elderly people. Methods The screening programme targeted colorectal cancer in old people was conducted on 1 100 participants aged 60-92 years (mean was 81.3 years). All subjects received OC-Hemodia and COB examinations. Positive subjects were examined by colonoscopy and/or barium enema. Out of 1 100 subjects 1 046 were followed up in the next five years. Results Among 1 100 subjects, 231 subjects (21.0%) were positive by OC-Hemodia test and 159 subjects (14.5%) were positive by COB. There was a significant difference in positive rate between OC-Hemodia test and COB (P

Objective To explore the therapeutic effect and possible mechanisms of Xuebijing injection for treatment of acute pulmonary thromboembolism(APE)in rabbits. Methods Twenty-four New Zealand rabbits with big ears were randomly divided into control group,model group and Xuebijing group(each n=8). The model of APE was performed by re-infusing of autologous blood clots. After the model was established,in the rabbits of the Xuebijing group,Xuebijing injection 1 mL/kg was injected into the ear marginal vein,twice daily for consecutive 7 days. In the model and control groups,the same amount of normal saline was given via the same route. The changes in plasma levels of B-type natriuretic peptide(BNP)were detected by enzyme-linked immunosorbent assay(ELISA) at the time points of 8 hours and 1,3,5,7 days,and after 7 days the lungs of the rabbits were removed for the lung histopathological examination. Results The BNP levels of the model group and Xuebijing group were significantly higher compared with the level of control group at 8 hours after embolization,the model group being the most significant(all P<0.05). On day 3,the BNP levels of the model group and Xuebijing group reached their peaks,and then gradually decreased,but the level of model group was significantly higher than that of Xuebijing group(ng/L:581.1±144.6 vs. 251.5±22.5,P<0.05). On day 7,the BNP level of Xuebijing group fell to the level of the control group(ng/L:78.5±9.4 vs. 50.9±11.5),the level of the model group also decreased,but it was still much higher than that of the Xuebijing group(ng/L:214.1±47.6 vs. 78.5±9.4,P<0.05). Histopathological examination showed Xuebijing could significantly reduce the infarct size with mild inflammatory cell infiltration,but the infarct size of the model group was obviously bigger than that of the Xuebijing group and accompanied by relatively more inflammatory cell infiltration. Conclusion Xuebijing injection can significantly reduce the plasma BNP level of the rabbits with pulmonary embolism,and in the mean time ameliorate the degree of inflammatory infiltration in the lung tissue and infarct.

Objective To observe the osteogenic and/or chondrogenic differentiation potential of cartilage endplate derived stem cells ( CESCs) .Methods The cartilage endplate ( CEP) was obtained from 5 patients who underwent posterior discectomy procedure for the lumber degenerative disease.The agarose culture was used to select CESCs.The expanded CESCs were injected into the hydroxyapatite ceram-ic(HA) and incubated for 7 days.After that,the HA contained CESCs were implanted into the subcutaneous of nude mouse for 7 weeks.Then the HA was obtained and sliced for the specific stain and immunohistochemisty.The expanded CESCs were injected into the degenerated in-tervertebral disk of the rabbit for 16 weeks.Then MRI was used to detect the repair of the degenerated disc.Results The metachromasia of toluidin blue indicated that cartilage matrix was formed in the vacant space of HA.Mallory stain indicated that there was bone matrix was formed.The results of immunohistochemistry indicated that there was more collageⅠand collageⅩ,and a small quantity of collage Ⅱ.MRI results indicated that the CESCs could be induced into the chondrocyte-like cells in the rabbit intervertebral disk.Conclusion The CESCs could be induced into the bone/cartilage in vivo.

OBJECTIVETo investigate the result of hip arthroplasty for failed internal fixation of femoral neck fractures.

METHODSFrom June 2007 to January 2014, 29 cases who underwent hip arthroplasty for failed of internal fixation of femoral neck fractures were reviewed. There were 12 males and 17 females. The mean age was 60.3 years (ranged 43 to 83 years) at the time of the fracture. Left hip was in 16 cases, right hip was in 13 cases. The average interval from fracture to arthroplasty was 23.3 months (ranged, 3 to 48 months).

RESULTSAll of 29 cases were performed total hip arthroplasty. There were 20 cases of cementless cup,7 cases of cementless cup with bone graft, 2 cases of cemented cup with bone graft; 13 cases of cementless stem, 16 cases of cemented stem. There were no complications occurred such as intraoperative fracture of the greater trochanter. The average operative time was (115 ± 38) minutes,the mean intraoperative blood loss was (420 ± 175) ml, the average postoperative drainage volume (240 ± 119) ml, intraoperative blood transfusion was (200 ± 220) ml, intraoperative fluid volume was (2,200 ± 400) ml, the average postoperative blood transfusion was (300 ± 200) ml. There was 1 case get postoperative dislocation. All patients were followed up for 14.7 months in average (ranged, 5 to 24 months). There was no revision for mechanical failure. Harris Hip Score significantly was improved from 51.1 ± 7.5 before the conversion to 88.5 ± 6.4 points at the final follow-up.

CONCLUSIONThe effect of the hip replacement for patients with failed internal fixation of femoral neck fractures was confirmed. This method can shorten the time on the bed and reduce the complications. It benefits the patients earlier functional recovery, but it must control operation indication. The long term efficacy is necessary to further observation.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femoral Neck Fractures ; surgery ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Treatment Failure

Objective To investigate the long-term excessive drinking semen nitric oxide (NO) content in their sperm quality and spermatogenic cell apoptosis and infertility. Methods Nitrate reductase method was used to specific reduce the NO metabolites nitrate (NO3-) to nitrite (NO2-), which was used on behalf of the total NO level. Terminal deoxynucleotidyl transferase (TdT) and mediated nick end labeling (TUNEL) method and binocular optical microscope were used to detect and observe the rate of apoptosis of spermatogenic cells and the morphological structure. The SQA-V sperm quality automatic analyzer was used to measure sperm quality. Results In not drinking semen fertility group, NO content was (54.81±11.45)μmol / L, the rate of spermatogenic cell apoptosis was (4.52±1.23)%, sperm motility was (80.24±0.17)%, energy a+b (78.32±0.12)%, deformity rate (5.30±0.13)%, and long-term excessive alcohol infertility C group was [(128.83±22.73)μmol/L,(17.34±2.53)%,(51.18±0.58)%,(21.45±0.26)%,(21.12±3.24)%] respectively. Compared to a very significant difference (t＝10.04,17.38,6.69,15.59,17.02,P＜0.01) . In long-term excessive drinking group, the levels of NO and spermatogenesis cell apoptosis rate was significantly positive correlated (r＝0.93,P＜0.01).Apoptosis of spermatogenic cell nucleus chromatin was condensed in the formation of crescent-shaped perinuclear, nuclear was cleavaged to form apoptotic bodies. Conclusions In the long-term excessive drinking semen, NO content and spermatogenic cell apoptosis rate was increased, the sperm had poor quality. The results showed that the long-term excessive drinking can cause germ cell apoptosis in male infertility and promote the body to overproduce NO, whichmay be one of the reasons for male infertility.

AIM:To compare the efficiency of fibrin glue to suture technique in pterygium surgery performed with limbal autograft under different methods of anesthesia.
METHODS: A prospective randomised clinical trial was carried out in 60 eyes of 55 patients operated for primary nasal pterygium, which were divided into two groups randomly: experimental group ( 30 eyes in 27 patients ) was under surface anesthesia ( oxybuprocaine ) and control group ( 30 eyes in 28 patients ) was under local anesthesia ( 20g/L lidocaine ). Autologous limbal graft taken from the superotemporal limbus was used to cover the sclera by a fibrin tissue adhesive after pterygium excision. Patients were followed up at least for 6mo. Time of operation, matching degree of graft and VAS score were mainly observed and recorded.
RESULTS: In experimental group the average surgery time was shorter (P=0. 008) and matching degree of graft ( 93%) was better than control group ( 83%) , the differences had statistical significance(P<0.05).
CONCLUSION: The surface anesthesia is enough when using fibrin glue for graft fixation in pterygium surgery, which will shorten surgery time and get better matching degree of graft.

Objective To provide reference for anti-infection drugs in Zunyi area by analyzing the etiological characteristics of children with severe pneumonia.Methods The sputum, throat swabs and serum of children with severe pneumonia in pediatric intensive care unit of our hospital from January 2014 to December 2015 were collected in this study.The pathogen species which caused severe pneumonia were detected and identified by the method of pathogen culture,and typical pathogens were detected by RT-PCR and indirect immunofluorescence.Results A total of 337 children with severe pneumonia were included,the pathogen positive rate was 86.65%(292/337).The rate of viral infection(37.32%)was the highest,followed by bacterial infection accounting for 28.42%,then the mixed infection accounting for 27.74%,and the mycoplasma pneumoniae infection accounting for 6.50%.The respiratory syncytial virus type B accounting for 28.44% was the most common in viral infection,and there were significant differences in age distribution(P<0.05),the children under 3-years-old,especially the infants under 1-year-old had the highest susceptibility.Viral infection had certain seasonality,compared with spring and winter,autumn and winter(November to April) had higher viral detection rate and the difference was statistically significant(x2=29.28,P=0.001).The escherichia coli was the most common in bacterial infection,accounting for 21.69%.Klebsiella pneumoniae,Escherichia coli were more common in neonates and infants under 1-year-old,Haemophilus influenzae occured in 1~3 years old children,Streptococcus pneumoniae infection could occur in any age.Conclusion Viral infection is the most common pathogen in children with severe pneumonia in Zunyi area followed by bacterial infection,which is more common in children under 1-year-old,and with the high incidence in autumn and winter.Bacterial infection is more common in children over 3 years of age.Bacterial and viral mixed infection is common in children under 3 years of age,especially in children under 1-year-old.

The potential of Rhizobium sp. N613 to produce the exopolysaccharide (REPS) was studied in this paper. Using an orthogonal design in a flask-shaker culture system, the fermentation medium and conditions of synthesizing REPS were optimized. Based on these results, the fermentation kinetic parameters were obtained in the batch fermentation with a 10L fermentor. The REPS yield of 11.31g/L was achieved by metabolic regulation during 40 h fed-batch fermentation. Transplanted tumor models of sarcoma 180 in mice were used to evaluate the anti-tumor effect. The result of anti-tumor activities showed that inhibition rate was 53.40%, when dose of REPS was 5mg/kg. These results indicate that REPS has the following properties: the short duration of fermentation, the high yield, the low cost, the effective immunocompetence and thickening. Thus, REPS has the value of development and application.

Objectives To investigate the relationship between fluorosis polymorphisms in collagen type Ⅰ alpha 2 (COL1A2) and osteocalcin (OC) gene, and serum calciotropic hormone levels. Methods The children between 8 and 12 years of age in Kaifeng and Tongxu cities of Henan Province were chosen to be the object of observation. Accoding to situation of dental fluorosis, they were divided into three groups: dental fluorosis group, non-dental fluorosis group from high fluoride areas, and control group form the control areas. The Pvu Ⅱ and Rsa Ⅰ markers of COL1A2 gene as well as HindⅢ marker of OC gene were genotyped by PCR-RFLP procedure. Calcitonin and osteocalcin levels in serum were measured using radioimmunassays. Results The frequency distribution of COL1A2 PvuⅡ genotype was pp 49.3%(37/75), Pp 32.0%(24/75), PP 18.7%(14/75) in children with fluorosis; pp 43.5% (30/69), Pp 52.2% (36/69), PP 4.3%(3/69) in children without fluorosis from high fluoride areas; and pp 43.8% (42/96), Pp 40.6% (39/96), PP 15.6% (15/96) in the children without fluorosis from control areas respectively. Childrens with the homozygous genotype PP of COL1A2 Pvu Ⅱ had a significantly increased risk of dental fluorosis(OR=4.85, 95%CI: 1.22-19.32) compared to children with the homozygous genotype pp in anendemic fluorosis area. The frequency distribution of COLIA2 Rsa Ⅰ genotype was rr 50.7% (38/75), Rr 36.0% (27/75), RR 13.3%(10/75) in children with fluorosis; rr 46.4%(32/69), Rr 46.4%(32/69), RR 7.2%(5/69) in children without fluorosis from high fluoride areas, and rr 45.8% (44/96), Rr 45.8% (44/96), RR 8.3% (8/96) in the children without fluorosis from control areas respectively. There were no significant differences in the three groups (P>0.05). The frequency distribution of OC Hind Ⅲ genotype was hh 48.0% (36/75), Hh 34.7% (26/75), HH 17.3% (13/75) in children with fluorosis; hh 43.5% (30/69), Hh 43.5% (30/69), HH 13.0% (9/69) in children without fluorosis from high fluoride areas, and hh 47.9%(46/96), Hh 40.6%(39/96), HH 11.5%(11/96) in children without fluorosis from control areas respectively. There were no significant differences in the three groups (P>0.05). Additionally, fluoride levels in urine and OC levels inserum were found to be significantly lower in controls from non-endemic areas compared to cases(P<0.05). However, the differences in urine fluoride and serum OC levels were not observed when cases were compared to controls from high fluoride areas(P>0.05). Conehlsions This study provides the evidence of an association between polymorphisms in the COL1A2 gene with dental fluorosis in populations exposed to high fluoride. There were no correlation between OC Hind Ⅲ genotype and the dental fluorosis.