Objective: To explore suitable concentration of recombinant human transforming growth factor β1 (rhTGF-β1) usage and study the effect of rhTGF-β1 on differentiation of dental pulp stem cells (DPSCs).Methods: DPSCs were isolated from the undiseased third molars of people aged 18-25 years and cultured according to instructions in vitro.Different concentrations (1 , 6 , 10 μg/L) of rhTGF-β1 were added to the culture medium to examine DPSCs proliferation by CCK-8 (cell counting kit-8) assay.The suitable concentration was then selected.For differentiation, the DPSCs were incubated for 7 or 14 days with rhTGF-β1 supplemented with osteo/odontoblastic induction medium containing 10 nmol/L dexamethasone, 10 mmol/L b-glycerophosphate, 50 g/L ascorbate phosphate, 10 nmol/L 1,25-dihydroxyvitamin D3 and 10% fetal bovine serum.The cells were then washed 3 times with phosphate-buffered saline and sonicated with 1%Triton X-100 for 30 minutes on ice.Cellular alkaline phosphatase (ALP) activity was assayed with p-nitrophenyl phosphate as the substrate.The enzyme activity was expressed as p-nitrophenyl produced per milligram of protein [bicinchoninic acid (BCA) protein assay kit].To examine mineral nodule formation, the cultured cells were fixed in 4% paraformaldehyde and washed in water, and the mineralization of the extracellular matrix was assayed by 1% alizarin red S staining and elution of staining was examined as optical density (D) under microplate reader.The mean difference was considered significant at 0.05 and 95% confidence interval.Results: The DPSCs had ty-pical fibroblast morphology and could form mineral nodules after being cultured with osteo/odontoblstic induction medium for 14 days.6 μg/L rhTGF-β1 significantly promoted the DPSCs proliferation on the 3rd and 5th days.After the incubation of osteo/odontoblastic induction medium, the DPSCs with the 6 μg/L rhTGF-β1 increased ALP activities compared with the control;D values in the 6 μg/L rhTGF-β1 group was 0.31±0.03, while the control group was 0.02±0.01(P<0.05).The total protein content in the 6 μg/L rhTGF-β1 group was (2 775.46±83.54) mg/L, and the control group was (1 432.20±110.83) mg/L (P<0.05).To eliminate the cells proliferation influence, relative ALP activities, which was defined as the total ALP divided by the total protein content, the 6μg/L rhTGF-β1 group was 6 times higher than the control group.Alizarin red S staining showed increased mineral nodule formation in the rhTGF-β1 group.The elution of staining under microplate reader also showed more optical density in the 6 μg/L rhTGF-β1-treated cells (0.83±0.02) than that in the control groups (0.55±0.05, P<0.05).Conclusion: 6 μg/L rhTGF-β1 could significantly promote DPSCs proliferation and odontoblastic differentiation in vitro.

AIM:To compare the therapeutic effect，adverse reactions and the costs between GM-CSF and G-CSF in treating leucopenia in chemotherapy of cancer.METHODS:Using pharmacoeconomic cost-effectiveness analysis，GM-CSF was compared with G-CSF in treatment of leucopenia in chemotherapy of cancer.RESULTS:The effective rate of GM-CSF was 80％ with an average cost of 1 008 yuan in a therapeutic course，the cost-effective ratio being12.6，and that of G-CSF was 85.7％ with an average cost of 2 304 yuan，the cost-effective ratio being 26.88.CONCLUSION:GM-CSF can effectively treat leucopenia in chemotherapy of cancer，and its cost-effective ratio ia superior to that of G-CSF.GM-CSF is worthy to be used clinically.

The incidence of venous thromboembolism after spinal cord injury is high, and its diagnosis and prevention received much attention in clinical research in recent years. Because of the special characteristics, the prevention of venous thrombosis should be combined with basic prevention, physical prevention and drug prevention. According to the different situations of venous thromboembolism in patients with spinal cord injury, the prevention methods need to adjust to carry out multidisciplinary comprehensive treatment.

Objective To observe the intracranial tumorigenicity situation of lung cancer cell strain and ovarian cell strain by injecting them into the female nude mice with different approaches and to analyze the possible mechanism for scarce brain metastasis of ovarian cancer to provide a certain thinking for the treatment of brain metastasis of lung cancer.Methods Ovarian cancer cell strain Skov3 and lung cancer cell strain A549 were injected into female nude mice respectively through tail veins,abdominal cavity,common carotid artery and brain tissue.There were 16 nude mice in each approach.All the nude mice were killed 4-6 weeks later.Then the brain,lungs,kidneys,spleen,liver,fallopian tube,ovary and abdominal tumor mass were taken by HE staining for observing the cerebral tumorigenicity situation.Results By the caudal vein injection,there was no brain metastasis in the Skov3 strain group,while in the A549 strain group,2 cases of brain metastasis occurred.By the aabdominal cavity injection,there was no brain metastasis in the Skov3 strain group,but 2 cases of brain metastasis occurred in the A549 strain group.By the common ca rotid artery injection,there was no brain metastasis in the Skov3 strain group,but 8 cases of brain metastases occurred in the A549 group.The intracranial tumorigenicity of Skov3 strain and A549 strain had statistical difference among these 3 kinds of extracranial injection approaches(P＜0.01).By intracranial direct injection approach,14 cases of brain tumorigenicity occurred in the Skov3 strain group,while 10 cases of brain tumorigenicity occurred in the A549 strain group,the difference had no statistical difference.Conclusion Blood brain barrier maybe plays an important role in preventing ovarian cancer cells from metastasizing into brain.

Objective:To understand the recognition status of cancer pain in basic medical staff from small towns to provide the basis for the improvement of cancer pain management in these areas. Methods:The medical staff of Hengjiang Town and subordinate villages was selected. The study area is situated in southwest China. Centralized questionnaires regarding cancer pain were collected and analyzed. A program and education of cancer pain were provided for these medical workers. Results: The medical staff from Hengjiang asserted that only 17%of cancer pain patients receive treatments. Approximately 70%of the medical staff did not consider the popularization and explanation of cancer pain treatment in their patients. Approximately 64%of the medical staff was not familiar with standardized cancer pain control, 87%did not believe that narcotics could suffice the need of patients, and 44%did not participate in the training for cancer pain control. Conclusion: The medical staff in Hengjiang possesses less knowledge on the importance of cancer pain. Hence, further training is necessary. The specific management of cancer pain as a part of community chronic diseases is mandatory.

Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007–2008, 28 human SFGR isolates and 34 rat SFGR isolates including 15 isolates from Rattus fulvescens, 5 isolates from R. edwardsi, 7 isolates from Callosciurus erythraeus roberti and 7 isolates from Dremomys rufigenis) were obtained from L929 cell culture. Previous research indicated that the 62 strains of SFGR mentioned above shared not only the same serophenotype but also 100% of identity sequences of 16S rRNA, gltA, ompA, groEL and 17KD, which enabled us to apply multispacer typing (MST) to the 62 SFGR isolates in the study. Six primer pairs, which were used for typing of Rickettsia rickettsii and Rickettsia conorii, were chosen, and the results exhibited greater nucleotide polymorphisms among the 62 isolates tested. A total of 48 distinct genotypes were identified. The dominant genotype, represented by h3 isolates, accounted for 21.7% (13/60) of the isolates tested, and the remaining 47 genotypes were all unique. Phylogenetic analysis showed that all the 48 genotypes could be classified in the same clade, while the genetically related strain, R. heilongjiangensis, was close but not the same as the cluster. We concluded that the genetically diverse of spotted fever group rickettsiae strains are endemic in Chengmai County, Hainan Province, China.

Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007—2008, 28 human SFGR isolates and 34 rat SFGR isolates including 15 isolates from Rattus fulvescens, 5 isolates from R.edwardsi, 7 isolates from Callosciurus erythraeus roberti and 7 isolates from Dremomys rufigenis) were obtained from L929 cell culture. Previous research indicated that the 62 strains of SFGR mentioned above shared not only the same serophenotype but also 100% of identity sequences of 16S rRNA, gltA, ompA, groEL and 17KD, which enabled us to apply multispacer typing (MST) to the 62 SFGR isolates in the study. Six primer pairs, which were used for typing of Rickettsia rickettsii and Rickettsia conorii, were chosen, and the results exhibited greater nucleotide polymorphisms among the 62 isolates tested. A total of 48 distinct genotypes were identified. The dominant genotype, represented by h3 isolates, accounted for 21.7% (13/60) of the isolates tested, and the remaining 47 genotypes were all unique. Phylogenetic analysis showed that all the 48 genotypes could be classified in the same clade, while the genetically related strain, R.heilongjiangensis, was close but not the same as the cluster. We concluded that the genetically diverse of spotted fever group rickettsiae strains are endemic in Chengmai County, Hainan Province, China.

Objective To evaluate the influence of ultrasound-mediated contrast agent microbubbles carrying kallidinogenase targeted therapy on neurogenesis and angiogenesis after experimental acute cerebral infarction.Methods Kallidinogenase-loaded microbubbles were prepared using mechanical shaking method.Middle cerebral artery occlusion (MCAO) model was established in male Wistar rats by sutureoccluded method.MCAO rats (n =120) were randomly divided into 5 groups:ultrasound-mediated kallidinogenase-loaded microbubbles group (group 1),ultrasound-mediated microbubbles group (group 2),ultrasound-mediated saline group (group 3),kallidinogenase group (group 4),saline group (group 5).Medication was given through tail vein daily for 2-6 consecutive days starting 24 h after MCAO.Ultrasound groups (groups 1,2 and 3) were given 2 MHz pulse ultrasonic irradiation on the ischemia lateral skull for 10 min after injection.Cell proliferation was examined using 5'-bromo-2'-deoxyuridine (BrdU,50 mg/kg).Infarction volume and neurological function were evaluated on the 3rd,7th day after MCAO respectively.Doublecortin (DCX) + cells in the subventricular zone and laminin + in the peri-infarction region were observed at the same time.Results The number of DCX + cells in the subventricular zone (under 20 × ocular) of groups 1-5 was 251.8 ± 13.1,125.7 ± 11.6,130.2 ± 13.7,234.5 ± 12.4 and 123.7 ± 10.0 respectively.The percentage of laminin + cells in the peri-infarction region (× 10 objective) of groups 1-5 was 10.0％ ± 0.8％,5.2％ ± 0.7％,5.0％ ± 1.0％,8.0％ ± 1.8％ and 5.0％ ± 0.9％ respectively.The number of DCX + cells in the subventricular zone and laminin + cells in the peri-infarction region of the group 1 and the group 4 was significantly more on the 7th day after MCAO compared with those of the other three groups (DCX:t values of group 1 vs groups 2,3,5 were 17.88,17.17 and 18.16,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 15.42,14.78 and 15.70,all P ＜0.01.Laminin:t values of group 1 vs groups 2,3,5 were 7.01,6.71 and 7.11,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 4.23,3.94 and 4.33,all P ＜0.01).Moreover,the number of DCX+ cells in the subventricular zone (t =2.46,P ＜ 0.05) and laminin + cells in the peri-infarction region (t =2.78,P ＜ 0.05) of the group 1 was much more than those of the group 4.Neurologic scores on the 7th day of groups 1-5 were 1.00 (0.75,1.25),2.0 (2.00,3.00),2.0 (1.00,2.00),1.5 (0.75,2.00),2.0 (2.00,2.50).Compared with other four groups,group 1 showed better functional improvement after stroke (U values of group 1 vs groups 2-5 were 2.0,4.0,7.5 and 2.5,all P ＜ 0.05).While there was no significant difference in infarction volume among five groups at all the time points after MCAO.Conclusions Our study demonstrates ultrasound-mediated kallidinogenase-loaded contrast agent microbubbles targeted therapy promotes neuroblasts proliferation and vascular regeneration compared with mediated and non-medicine-loaded microbubbles therapy,which attributes to functional improvement after MCAO.Therefore,ultrasound-mediated ultrasound contrast agent microbubbles carrying drugs targeted therapy may have a perspective on ischemic stroke.

Objectiveto investigate serum leptin concentration and lipdid levels in patients with esophageal squamous cell carcinoma and their clinical significance. MethodsBlood samples were collected from 47 patients with esophageal cancer, 20 healthy control subjects. Serum leptin was measured by ELISA and serum lipids was measured by biochemistry auto-analyzer. ResultsThe serum leptin concentration and LP α levels in patients group were significantly higher than those in control group[(13.09±5.94) ng/ml vs (7.584±4.15) ng/ml,(216.50±752.52) ng/ml vs(87.85±10.73) mg/L](P=0.001).However,the TG concentration was significantly lower than control group [(1.23±0.46) mmol/L vs (1.58±0.07) mmol/L]( P=0.001 ).Multiple logistic regression analysis revealed statistically significant association between serum leptin levels and esophageal squamous cell carcinoma incident(ORLeptm =1.442,95％ CI 1.094-1.848). ConclusionThe increased serum leptin levels maybe the risk factors of esophageal cancer,elevated LP α levels in patients might be a compensatory reactions.

Objective To compare the effect of 89Sr radionuclide therapy and radiotherapy in the treatment of metastatic cancer pain of vertebral column. Methods 80 patients with cancer pain of vertebral metastasis in the second People's hospital of Yibin from April 2015 to April 2017, were randomly divided into the control group treated by radiotherapy treatment (n=40) and the the observation group treated by 89Sr radionuclide therapy(n=40), and the effect of treatment were compared between two groups. Results In the observation group, the total effective rate of pain response in patients with metastatic cancer pain of vertebral column was 85.0％, and the control group was 82.5％. There was no significant difference between two groups. The onset time of treatment in the observation group was (6.5±1.7)d, significantly shorter than that of the control group (12.9±2.6)d, and the difference was statistically significance (P<0.05). Conclusion The application of 89Sr radionuclide therapy in the treatment of vertebral metastatic pain is equivalent to the radiotherapy in improving the pain response, but the efficacy could be achieved in a short period of time, so it is worth popularizing widely in clinical practice.