Objective　To approach the relationship betwe en glycosaminoglycan metabolism in cartilages and pathogenesis of KBD.Methods　Rhesus monkey was fed with grains and water from KBD endemic area for 18 months to produce the animal model with KBD . The glyc osaminoglycans in the monkey cartilage were extracted by the improved Dish method of Bitter. Purified glycosaminoglycans were digested with chondroitinaseABC, and the enzymatic digests were analyzed by HPLC. Results　Comparing with those of the control, the glycos aminoglycans in the head of femur, tibia plateau and costal cartilage from the Rhesus monkey fed with grains and water from KBD endemic area were undersulfated . Decreased unsaturated 4-sulphated disaccharide (△Di-4S) from the glycosa minoglycans in the head of femur and tibia plateau and decreased unsaturated 6- sulphated disaccharide (△Di-6S) from the glycosaminoglycans in the costal cart ilage were discovered.Conclusions　Detrimental factors in grains and water from KBD endemic area cause undersulfate of the cartilages glycosaminoglycans from Rhesus monkey. The glycosaminoglycans changes have a direct bearing on the patho logical alterations in morphology on the cartilages from the animal model with K BD.

Objective To evaluate the relationship between cold hyperalgesia and trafficking of transient receptor potential melastatin 8 (TRPM8) to cell membrane in the dorsal root ganglion (DRG) of rats with neuropathic pain (NP).Methods Ninety-six healthy male Sprague-Dawley rats,aged 10-12 weeks,weighing 250-280 g,were divided into sham operation group (S group,n=48) and NP group (n =48) using a random number table.NP was produced by chronic constriction injury to the sciatic nerve.The number of paw lifts on the cold plate and mechanical paw withdrawal threshold (MWT) were measured on 1 day before operation and 1,4,7,10 and 14 days after operation.Rats were sacrificed after behavioral testing,and ipsilateral DRGs of the lumbar segment (L46) were dissected tor detection of the expression of TRPM8 in total and membrane proteins by Western blot,and the ratio of TRPM8 expression in the membrane protein to that in the total protein (m/t ratio) was calculated.Results Compared with group S,the number of paw lifts on the cold plate was significantly increased,the MWT was decreased,the expression of TRPM8 in total and membrane proteins was up-regulated,and m/t ratio was increased on postoperative days 4,7,10 and 14 in group NP (P＜0.05 or 0.01).In group NP,the number of paw lifts on the cold plate was gradually increased with the prolongation of time after operation and reached the peak on postoperative day 10,maintaining at the peak until postoperative day 14;the MWT was gradually decreased and reached the lowest level on postoperative day 10,maintaining at the lowest level until postoperative day 14;the expression of TRPM8 in total and membrane proteins and m/t ratio were gradually increased with the prolongation of time after operation and reached the peak on postoperative day 10,maintaining at the peak level until postoperative day 14 (P＜0.01).Conclusion The mechanism underlying the development of cold hyperalgesia is related to enhanced trafficking of TRPM8 to cell membrane in DRGs of rats with NP.

Objective To investigate the gene mutation of epithelial growth factor receptor(EGFR) and the expressions of hu‐man epiderma1 growth factor receptor 2(HER‐2) and vascular endothelial growth factor(VEGF) in lung adenocarcinoma and their relationships with the clinical pathological factors .Methods ARMS‐PCR method was used to detect the gene mutation of EGFR in 49 lung adenocarcinoma specimens and 10 normal tissue specimens .Immunohistochemical method was also used to detect the ex‐pressions of the HER‐2 and VEGF in them .Results (1) The mutation rates of EGFR and the positive rates of HER‐2 and VEGF in lung adenocarcinoma were significantly higher than those in the normal tissues(P< 0 .01) .(2) The occurs of EGFR mutation were correlated with sex and the smoking habit(P<0 .05);but not correlated with age ,the size of tumor ,differential ,lymph nodes metastasis ,TNM stages and pleural invasion(P>0 .05) .The HER‐2 and VEGF protein expressions in lung adenocarcinoma were correlated with differential ,the size of tumor ,TNM stages ,lymph nodes metastasis and pleural invasion(P<0 .05);but not correla‐ted with sex ,age ,and the smoking habit(P>0 .05) .(3) The expressions of HER‐2 and VEGF protein in the lung adenocarcinoma were positively correlated with each other(P<0 .01) .Conclusion EGFR mutation is closely related to the occurrence of lung ade‐nocarcinoma ,its high expressions in the women ,non smoking people have important clinical significance .HER‐2 and VEGF could promote the lung adenocarcinoma′s occurrence ,development and transfer .They could be used to evaluate the patients′prognosis ,and provide new molecular targeted therapy .

Objective To explore the effect of spinal decompression system on lumbar disc herniation. Methods 83 patients of lumbar disc herniation were divided into observation group (n=42) and control group (n=41). Both groups received massage therapy, while the observation group was treated with spinal decompression system in addition. The effect was evaluated with Visual Analogue Scale (VAS) and Japanese Orthopedic Association (JOA) assessment of low back pain. Results The VAS score decreased and the JOA score increased in both groups after treatment (P<0.05), and it was better in the observation group than in the control group (P<0.01). Conclusion Spinal decompression system could improve the therapeutic effect on lumbar disc herniation.

Objective To screen the arsenic content situation of drinking water in lower reaches of Yellow River and survey the amount of threatened people drinking high arsenic water and the condition of endemic arsenism.Methods Four counties of Yuncheng,Jiaxiang,Dongchangfu and Boxing were selected to colleft the water samples by CroOSS-sectional survey method.The water arsenic content wag determined by semi-quantitative rapid kit.All water samples having arsenic were re-determined by atomic fluorescence spectrometry.And the nurober of threatened people who drinking high arsenic water were investigated.Results In 4765 water wells screened,303 water samples had contained arsenic,arsenic content of 35 samples Was≥0.030 mg/L,12 samples were exceeding the international standard (arsenic content≥0.050 ms/L),they distributed in 3 counties of Dongchangfu,Yuncheng and Jiaxiang.The residents drinking water wells of arsenic content≥0.030 mg/L were surveyed by epidemiological investigation.And in the 28 villages 13 032 residents and 11 Bu8picious patient8 wlere checked out.Conclusion The wells with excesive water arsenic content are existing in the lower reaches of Yellow River and people suspicious of endemic arsenism need to be further identified.

Adult ; Chordoma ; pathology ; Female ; Humans ; Nose Neoplasms ; pathology

Objective To explore the clinical characteristics and treatment of Caroli′s disease in children.Methods The clinical data,laboratory examination and radiological feature of 8 children with Caroli′s disease between Feb.1998 and Dec.2007 were analyzed retrospectively.All children underwent CT and abdominal ultrasonogram.Results Five cases of the 8 children were male and 3 cases were female.The mean age was 6.3 years old.The cases′ history were from 5 days to 4 months.The clinical symptoms showed that 3 cases had hematemesis,5 cases had hepatosplenomegaly,and 1 case had fever and turbid urine.Of the total 8 cases,5 cases were hepatic fibrosis and liver cirrhosis and hepatosplenomegaly,3 cases were portal hypertension,and 1 case had cholangitis.The other 3 cases were simple types.One case had infantile polycystic kidney disease.Laboratory analysis revealed 2 cases had dysfunction of liver and 1 dysfunction of renal.The imaging characteristics showed multiplied irregular dilatation of the intrahepatic bile ducts in enlarged liver,with central dot sign on CT scan.One case presented enlarged gastroesophageal vein.The 8 cases undertook conservative treatment,with no surgery.Conclusions The symptoms of Caroli′s disease are highly variable.Caroli′s disease should be focused especially on children with abdominal pain and hepatomegaly.CT is important for diagnosis of Caroli′s disease at earlier stage.The disease can be conservatively treated,and(or) surgically operated.

Objective To explore the reliability of relevant electrocardiogram(ECG) indexes in evaluating isoprenaline(ISO)-induced rat acute ischemic myocardial injury and provide reference for future scientific applications of these models. Methods Seventy male Wistar rats were randomly equally assigned to ten groups according to their body weight: 5,10,20,40,80,160,320,640, 1280 and 2560 μg/kg dose groups. All rats were tail intravenously given corresponding doses of saline diluted isoprenaline according to their body weight. Standard limb Ⅰ , Ⅱ, Ⅲ-lead ECG of all rats were recorded before, immediately after and 1,24,and 72 hour after injection, respectively.Changes of heart rate, T-wave amplitude of Ⅱ -lead and Q-T interval were measured. Results Significant differences were found in heart rates, T-wave amplitudes and Q-T intervals at different time points(F = 15.03,11.28,13.64, all P ＜ 0.01 ), while differences among the ten ISO-dose groups were statistically insignificant (F= 1.45, 1.17,1.09, all P ＞ 0.05). No interaction between observation time and ISO dose was observed on heart rates, T-wave amplitudes and Q-T intervals(F= 0.79,0.82,0.59, all P ＞ 0.05). Immediately after injection of ISO, the heart rates were significantly increased compared with that of pre-injection in all groups(all P ＜ 0.05), of which 320 and 640μg/kg dose groups increased most significantly [(550 ± 47), (521 ± 43)times/min]. T-waves decreased significantly compared with that of pre-injection (all P ＜ 0.01 ), and 20 μg/kg dose and above groups decreased particularly evident, and partly inverted. Q-T intervals of rats in each group were significantly shorter than that of pre-injection(all P ＜ 0.01 ), and 320, 640, 1280 μg/kg groups shortened more pronounced[(0.070 ± 0.006),(0.072 ± 0.005), (0.068 ± 0.005)ms]. One hour after injection, the heart rate of rats in each group decreased,except 320 and 640 μg/kg dose groups[(518 ± 43), (487 ± 36)times/min], which were still higher than that of pre-treatment[(450 ± 40), (448 ± 51 )times/min, all P ＜ 0.05], the rest groups no longer had significant differences (all P ＞ 0.05). ECG T-wave in each group was significantly recovered compared with that of instantly medication (all P＜0.05), and 40 μg/kg dose and above groups recovered more than a big margin, but there were still differences compared with that of pre-treatment (P ＜0.05), while T-waves of 40 μg/kg dose and below groups had returned to the level of pre-treatment. Q-T interval in each group had varying degrees of recovery, except 1280 and 2560 μg/kg dose groups[(0.080 ± 0.004), (0.076 ± 0.011 )ms]which were still less than that of pre-treatment[(0.086 ± 0.007),(0.085 ± 0.006)ms, all P ＜ 0.05], other groups had no significant difference compared with that of pre-treatment (all P ＞ 0.05). Twenty-four hours after injection of ISO, the heart rates of 1280 and 2560 μg/kg dose groups [(389 ± 31 ), (398 ± 23)times/min]decreased significantly compared with that of pre-treatment[(427 ± 43), (438 ±26)times/min, all P ＜ 0.05], while other groups had returned to the level of pre-treatment. Seventy-two hours after injection of ISO, the heart rates, T-wave amplitudes and Q-T intervals of all doses groups had returned to the level of pre-treatment (all P ＞ 0.05). Conclusions There has no significant ST segment in the electrocardiogram of rat.Isoprenaline has an exact effect on shortening Q-T interval. T-wave amplitude and Q-T interval can be used as reliable indexes of ECG for assessment of this animal model.

Objective To study the lysozyme activity in Oncomelania hupensis and observe its inhibitory effect on bacterial growth.Methods Soft tissues of Oncomelania hupensis were initially homogenized and immersed in Tris-HCl-TritonX-114 buffer solution for 24 hours then the supernatant was collected after centrifugation at 10 000 × g for 10 minutes.The supernatant was incubated in a 37 ℃ water bath for 15 minutes and centrifuged again at 2000 × g for 10 minutes.The precipitate was put into ultrafiltration tube (relative retention molecular mass =3000) and centrifuged at 4 ℃,5 000 × g for 30 minute to obtain concentrated enzyme.The protein content,lysozyme activity and the antibacterial effect on Micrococcus lysodeikticus,Shigella dysenteriae,Staphylococcus aureus,Escherichia coli and Candida albicans were measured with bicinchonininc acid(BCA) method,turbidimetric method and agar diffusion (K-B) method,respectively.Results The antibacterial protein lysozyme was identified in gastropod protein concentration of the concentrated enzyme was 3.428 g/L.Average activity,total activity,and specific activity were (760 ± 120) × 103 U/L,(1520 ± 240) × 103 U/L and (221.70 ± 35.00)U/mg,respectively.The enzyme had produced exclusive inhibitory effects on growth of Micrococcus lysodeikticus and Shigella dysenteriae.Average inhibitory diameters were 10-12 and 12-15 mm,respectively.No inhibition zone was observed in saline control,Staphylococcus aureus,Escherichia coli and Candida albicans.Conclusions Lysozyme can be extracted from soft tissues of Oncomelania hupensis with Tris-HCl-TritonX-114 buffer solution,and the enzyme has inhibitory effect on growth of Micrococcus lysodeikticus and Shigella dysenteriae but has no antibacterial effect on Staphylococcus aureus,Escherichia coli and Candida albicans.

Objective To prepare biphasic calcium phosphate/polyvinyl alcohol scaffolds by 3D printing at room temperature and explore the effect of 3D scaffolds on in vitro osteogenic differentiation of the bone marrow mesenchymal stem cells (BMSCs).Methods After biphasic calcium phosphate and polyvinyl alcohol solutions were mixed,the biphasic calcium phosphate/polyvinyl alcohol composite scaffolds were prepared by room temperature 3D printing combined with freeze drying technique.Non-printing scaffolds were prepared by injection molding.The surface microstructure,porosity,elastic modulus and hydrophilicity of the 2 sorts of scaffolds were measured.The cytological experiments were carried out in 3 groups (n =3):printed scaffold group,non-printed scaffold group and blank control group (no scaffold).After the BMSCs were seeded onto the scaffolds for 7 and 14 days,the 3 groups were compared in terms of cellular proliferation,alkaline phosphatase activity and expression levels of osteogenesis-related genes.Results 3D composite scaffolds with controllable pore size and porosity were prepared successfully,with an average porosity of 59.6％ ± 3.6％ and an average elastic modulus of 429.3 ± 54.3 kPa.After culture for 7 and 14 days,the cellular absorbance values in the printed scaffold group (0.987 ± 0.047 and 1.497 ± 0.076) were significantly higher than those in the non-printed scaffold group (0.767 ±0.063 and 1.181 ±0.098) (P ＜ 0.05) which were in turn significantly higher than those in the blank control group (0.532 ±0.046 and 0.895 ± 0.062) (P ＜ 0.05).After culture for 7 and 14 days,the ALP activity and expression levels of osteogenesis-related genes in the printed and non-printed scaffold groups showed no significant between-group differences (P ＞ 0.05),but were significantly higher than those in the blank control group (P ＜ 0.05).Conclusions Tissue-engineered composite biphasic calcium phosphate/polyvinyl alcohol scaffolds with controllable pore size and good connectivity can be prepared by freeze-drying and room temperature 3D printing techniques.Co-culture of the scaffolds and BMSCs in vitro promotes adhesion,proliferation and osteogenic differentiation of the cells.