Objective:To investigate the effect of genistein,a soybean-derived isoflavone,on thestimulating effect on bone resorption of IL-1?.Methods:The osteoclasts(OCs)were isolated with themethods of Yu Shifeng.The rat calvaria were cultured as an organ.The cells in the experiment weregrown in four respectively:Control(without genistein or IL-1?),10~(-6) mol/L genistein,10 ?g/L IL-1?and 10~(-6) mol/L genistein+10 ?g/L IL-1?.The area of bone resorption,the concentration of Ca~(2+) in thesupernatant liquid of OCs cultures and mice calvaria were tested.The contents of acid phosphatase(ACP)were also examined by biochemistry method.The index of bone resorption was counted as the ra-tio of the experiment average and control ones,which indicated the increase in bone resorption when itwas above 1.0.Results:The area of bone resorption of 10~(-6) mol/L genistein+10 ?g/L IL-1? increasedcompared with that of 10~(-6) mol/L genistein,while the concentration of Ca~(2+) in the supernatant liquid ofOCs cultures decreased significantly.The index of bone resorption of 10~(-6) mol/L genistein+10 ?g/L IL-1? lied between 10~(-6) mol/L genistein and 10 ?g/L IL-1?.In the organ culture,there was no differencein the content of ACP among all the groups,The index of bone resorption of 10~(-6) mol/L genistein+10?g/L IL-1? was below that of 10?g/L IL-1?,but both were above 1.0.The index of bone resorptionwas below 1.0 in the group of 10~(-6) mol/L genistein.Conclusion:Genistein can suppress obviously thebone resorption simulated by single IL-1?.

Objective To investigate the expressions of TESTIN gene and Caspase-3 protein and thier relations with the clinicopathological features and prognosis of esophageal squamous cell carcinomas(ESCC). Methods The expressions of TESTIN and Caspase-3 in 50 ESCC tissues and paracancerous tissues (＞5 cm apart form the ESCC tissue) were detected by immunohistochemistry.The expression of TESTIN in 65 matched-pairs of ESCC tissues was detected by Western blotting and RT-PCR. The association of TESTIN and Caspase-3 with clinicopathological features and prognosis of ESCC were analyzed. Results The immunohistochemistry examination showed that the positive rates of TESTIN protein [30. 0% (15/50)] and Caspase-3 protein [24.0% ( 12/50)] were significantly lower in ESCC tissues than those in paracancerous tissues [(84. 0% (42/50) and 94. 0% (47/50),respectively, P＜0.01)]. The mRNA level of TESTIN was down-regulated in ESCC tissues (P＜0.05). Whereas the protein level of TESTIN was down-regulated in 45 (69.2%) of 65 ESCC tissues in comparison with paracancerous tissues. TESTIN expression was positively correlated with the differentiation of ESCC, but not associated with gender, age, tumor size, TNM stage and lymph node metastasis. There was a positive correlation between TESTIN and Caspase-3 in protein expressions (P＜0. 05). Patients with negative expression of TESTIN had lower survival rate compared to those with positive expression (P＜0. 05). Conclusions The positive relation between low-expression of TESTIN and Caspase-3 implicates that both are involved in the development of esophageal squamous cell carcinogenesis, and TESTIN might be a novel ESCC marker with prognostic significance.

Objective To evaluate the effect of 36-neuro-motorial examination on early diagnosis of children with cerebral palsy within 1 year-old.Methods36-neuro-motorial examination was analysed in 210 children with cerebral palsy from 2 to 12 months. Results4-29 items reflecting the abnormity of reflex, muscle tone, posture and motion were observeed in patients with cerebral palsy.If these abnormities were laid according as frequency among 2-6 months,reflex abnormities are knee tendon reflex, palmar grasp reflex,Babinski sign,asymmetrical tonic neck reflex,ankle clonus,sitting equilibrium reflex,stepping reflex,crossed extension reflex,tonic labyrinthine reflex,trunk incurvation reflex,Moro reflex; posture abnormities are head to back≥15°,from supine to the side position, axillar suspension reflex,posterior neck fovea≥1cm in the supine position,landau reflex,Vojta reflex,traction reflex,tonic torticollis,abnormal spontaneous posture;muscle tone abnormities are foot dorsiflexion angle,scarf sign,tonic palmar grasp,to up the foot heel≥30° in the stand position,thumb decussation to the palm,poples angle,adductor angle,heel-ear angle; others are head cycle≤-2s, squint, active movement attenuation or abnormity, auditory abnormity, visual abnormity, ophthalmodonesis, epilepsy.Conclusions 36-neuro-motorial examination is effective for early diagnosis of cerebral palsy within 1 year.

Objective To detect the type of gene mutation of thalassemia in Kunming city.Methods Sixty-three cases highly suspec-tive of thalassemia were determined with the methods of ploymerase chain reaction(PCR) and reverse dot blot(RDB) for the type of gene mutation.Results According to gene analysis,35 cases were final diagnosed from 63 cases suspective of thalassemia.Among the total,4 cases were gene deficiency ?-thalassemia,and 30 cases were gene deficiency ?-thalassemia,and there was 1 case both ?-thalassemia and ?-thalassemia.There were 9 types of gene mutation with 15 gene combinations in 35 samples.The main type of ?-thalassemia was--SEA/??,there were 6 types with 11 gene combinations from the types of genes of ?-thalassemia,the highest incidence of gene mutation was 17 site,including 17 site homozygote,heterozygote and double heterozygote.Conclusions The thalassemia invasion of Yunnan has its characters,and it is valuable to launch further research.In the same patient,there are ?-thalassemia and ?-thalassemia,it signifies that those 2 types should be diagnosed in the same time,to prevent missed diagnosis.

Objective To investigate the effects of maternal long-term exposure to low-dose trichlorfon on the serum paraoxonase (PON) activity of pregnant mice and development of embryos. Methods Female ICR mice (n=120) were randomly divided into control group and trichlorfon groups of different doses,and were managed by intragastric injection with trichlorfon of 0,2,10 and 50mg/kg,respectively. All the mice were managed once a day for a consecutive of 27 days,and were subjected to mating. The pregnant mice were continued to be managed with trichlorfon for 3 days,and were sacrificed on day 3 of gestation. The serum PON and acetylcholinesterase (AchE) activities were detected,and the development of embryos was evaluated. Results The serum PON activity of 2,10 and 50mg/kg trichlorfon group were (14.15?1.22),(12.78?1.80) and (10.45?1.95)IU/mL,respectively,and that of 50mg/kg trichlorfon group was significantly lower than that of control group [(13.37?2.31)IU/mL] (P0.05),while the the percentage of abnormal embryos of 50mg/kg trichlorfon group had an increased tendency. Conclusion Long-term exposure to low-dose trichlorfon can inhibit serum PON and AchE activity in pregnant mice without obvious effect on the development of embryos.

OBJECTIVETo evaluate the accuracy and safety of pedicle screw insertion with the aid of novel patient-specific drill-guide templates in scoliosis cases.

METHODSTen patients with scoliosis were selected to participate in the research (the observation group) from December 2013 to December 2014. The data was obtained from CT scanning, and put into the computer to perform reconstruction of spine, simulation of pedicle screw insertion, and design of patient-specific drill-guide templates with software. The templates were made with rapid prototyping technique. After sterilization, the templates were used to aid the pedicle screw insertion intraoperatively. The blood loss, operation duration, change of creatinine level pre- and post-operation, and complications related to pedicle screw insertion were recorded. The location of pedicle screws were graded so as to evaluate the accuracy. A comparative study was then performed with the data of ten scoliosis cases operated with free-hand method during the same period (control group). There were 5 cases of idiopathic scoliosis and 5 cases of congenital scoliosis in the observation group, including 3 males and 7 females. Their average age was 11.9 years old (ranged, 4 to 18 years old), and the average Cobb angle of main curve was 54.9° (ranged, 42.1° to 78.4°). There were also 5 cases of idiopathic scoliosis and 5 cases of congenital scoliosis in the control group,including 2 males and 8 females. Their average age was 12.6 years old (ranged, 6 to 17 years old), and the average Cobb angle of main curve was 56.6° (ranged, 38.2° to 93.4°).

RESULTSA total of 167 pedicle screws were inserted intraoperatively, with 138 screws (82.6%) in grade I, 26 screws (15.0%) in grade II, 4 screws in grade III (2.4%), but no screws in grade IV according to the CT image. There were 29 (17.4%) screws perforated, and 163 (97.6%) screws could be accepted. In the control group, a total of 165 pedicle screws were inserted intraoperatively, with 98 screws (59.4%) in grade I, 39 screws (23.6%) in grade II, 21 screws in grade III (12.7%), and 7 screws in grade IV (4.2%). There were 67 (40.6%) screws perforated, and 137 (83.0%) screws could be accepted. The grade distribution of screw position, ratio of perforated and accepted screws were significantly different between the two groups respectively (Z=-5.013, P=0.000; χ2=9.347, P=0.002; χ2=20.242, P=0.000). The correction rate of Cobb angle were (74.1±10.0)% vs (69.7±17.6)%; blood loss were (455±447) ml vs (415±389) ml; operation duration were (163.5±53.7) min vs (164.0±48.7) min; and the changes of creatinine level pre- and post-operatively were (-5.3±3.2) μmol/L vs (-3.4±3.1) μmol/L; all above data had no significant differences respectively (t=0.696, P=0.496; t=0.214, P=0.833; t=0.022, P=0.983; t=1.375, P=0.192). There were no complications related to pedicle screw insertion in each group.

CONCLUSIONThe novel patient-specific drill guide template can be used to assist the insertion of pedicle screws in scoliosis cases with much higher accuracy than that of freehand method and fair safety.

Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Male ; Pedicle Screws ; Scoliosis ; surgery ; Tomography, X-Ray Computed

Objective To explore the in vitro spermostatic effects and the mechanisms of ceritinib,a novel candidate from the active compound pools previously screened for the regulation of sperm function.Methods The vigor sperm of human and mouse were incubated with ceritinib for 20seconds,and the sperm motility was evaluated by computer assistant sperm analysis (CASA).The integrity of the sperm plasma membrane and the survival ratio of sperm was assessed by hypo-osmotic swelling (HOS) assay and SYBR-14/PI staining.The damage of sperm plasma membrane was detected by electron microscope.The cytotoxicity of ceritinib to VK2/E6E7,End1/E6E7 and Ect1/E6E7 cells was measured by CCK-8 assay and fluorescent staining.Results The minimal effective concentration (MEC) of ceritinib to (5-10) × 106/mL human sperm within 20 seconds was (74.87 ± 31.46)μmol/L,which was significantly lower than the MEC of nonoxynol-9 (219.75 ± 20.89) μmol/L measured in the same condition.The time-dose related spermostatic effects of ceritinib were measured by CASA system.Ceritinib was able to damage sperm membrane and caused sperm death with HOS and SYBR-14/PI staining.With electron microscopy,the sperm membrane was observed to be ruptured,and the heads and tails of sperm were separated by ceritinib.Ceritinib was also able to damage the epithelial cells.Conclusions Ceritinib has acute spermostatic effects and potential contraceptive effects.

Objective To study the cell death in macrophages (THP-1) stimulated with different agonists (H37Rv-PPD or BCG-PPD) and to investigate the relationship between Toll like receptor (TLR)-2 and THP-1 apoptosis. Methods H37Rv-PPD and BCG-PPD were used to stimulate THP-1 cells for 3 h, 8 h, 15 h and 24 h, respectively with or without TLR-2 blockade. Cells were analyzed by flow cytometry to detect the TLR-2 expression. Annexin V staining and Hochest staining were performed to evaluate apoptosis. Results The apoptosis cells were increased when stimulated with BCG-PPD and the percentage was 30.2% at 24 h, which were confirmed by Hochest staining.However, the expression of TLR-2 did not increase simultaneously with percentage of 8.8% at 24 h.Nevertheless, most cells presented with necrosis form when stimulated with H37Rv-PPD and the expression of TLR-2 remained at high level with the percentage of 17.2% at 24 h, while the percent of apoptosis rate was only 7.7%. Under treatment of TLR-2 antibodies, the percentage of apoptosis decreased to 10.5% at 24 h of BCG-PPD stimulation and TLR-2 expressions were down-regulated to less than 3% at all time points; but after H37Rv-PPD stimulation, the percentage of apoptosis and TLR-2 expression did not changed obviously. Conclusions The attenuated BCG-PPD induces THP-1 apoptosis predominately, which is partially correlated with TLR-2 expression. While virulent H37Rv-PPD induces THP-1 necrosis predominately.

Objective To investigate the relationship between amnestic mild cognitive impairment and functional genes associated with hyperphosphorylated tau protein.Methods One hundred and sixteen amnestic mild cognitive impairment (aMCI) patients and 93 normal controls were recruited for the study.Multi-dimension neuropsychologic tests were used to assess the cognitive function extensively.MassARRAY and iPlex systems were used to measure candidate SNP polymorphisms,analyze genotypic,allelic or haplotypic distributions and their interaction with ApoE ε4 and the correlation with the cognitive function in the subjects.Results ( 1 ) The scores of neuropsychologic tests in memory domain ( Auditory Verbal Learning Test (AVLT)-first immediate recall,AVLT-second immediate recall,AVLT-second immediate recall,AVLT-5 minute delayed recall,AVLT-20 minute delayed recall,AVLT-recognition,Rey-Osterrich Comolex Test-delay) in aMCI patients ( 3.0 ( 0-7.0 ),5.0 ( 1.0-10.0),6.0 ( 1.0-11.0 ),4.0 (0-11.0),3.0(0-10.0),20.0(8.0-24.0),11.2 ±8.3) were significantly lower than those in the normal controls(4.0(0-9.0),7.0(2.0-11.0),9.0(3.0-12.0),8.0(0-12.0),8.0(0-12.0),22.0 (10.0-24.0),16.1±8.0) (Z=-3.592,-6.802,-6.408,-8.173,-8.533,-5.647 andt=4.216 respectively,all P ＜0.01 ) ; (2) Genotypic distributions of rs242562 GG in aMCI (7.826％ ) were significantly lower than those in normal controls (20.65％,OR =0.3525,95％ CI 0.1411-0.8807,P =0.024 98),however there were no differences in the genotypic,allelic or haplotypic distributions between aMCI patients and controls of glycogen synthase kinase-3β,cyclin dependent protein kinase-5,calcium and calmodulin-dependent protein kinase-Ⅱ,cell division cycle 2,dual-specificity tyrosine-phosphorylation regulated kinase 1A and low density lipoprotein receptor-related protein 6; (3) MAPT/STH rs242562 genotype was correlated with AVLT-immediate recall,AVLT-delayed recall,Rey-Osterrieth Complex Test,Rey-Osterrieth Complex Test-delayed recall and Clock Drawing Test (H =9.763,12.258,10.508,9.624,10.767,F =3.700,3.123 and H =6.591 respectively,all P ＜ 0.05 ) ; (4) There were no differences in the distributions of MAPT/STH rs242562 GG genotype and ApoE ε4 haplotype between aMCI patients and normal controls.Conclusions MAPT/STH rs242562 GG genotype decreases the genetic risk of aMCI,which might have important role in memory function in aMCI.The interaction between rs242562 GG and ApoE ε4 doesn' t affect the susceptibility to aMCI.

Objective To explore the mechanisms by which hydrogen sulfide (H2S) regulates collagen metabolism during hypoxic pulmonary vascular structural remodeling. Methods Wistar rats were randomly divided into control group (n=6), hypoxia group (n = 6) and hypoxia+NaHS group(n= 6) H2S content in plasma was measured with spectrophotometry. The expression of collagen I was detected by immunohistochemistry. The expression of tissue inhibitor of metalloproteinase-1(TIMP-1) mRNA was detected with situ hybridization Results Compared with hypoxia group, H2S content in plasma increased significantly and the expressions of collagen I and TIMP-1 mRNA in pulmonary arteries were down- regulated in hypoxia + NaHS group. The H2S content in plasma negatively correlated with TIMP-I mRNA expression. Conclusions H2S increases the degradation of collagen in small and media pulmonary arteries of rats under hypoxia by decreasing the synthesis of TIMP-1, and therefore, attenuats the collagen remodeling in pulmonary arteries of hypoxic rats.