Objective To investigate the significance of osteo-immunology related factor transforming growth factor-β1 (TGF-β1) and interleukin 6 (IL-6) in bone of rats with chronic fluorosis.Methods Thirty-six healthy SD rats were divided to three groups according to body weight with the method of random digits table.The rats of control were fed with tap water(NaF ＜ 1 mg/L) and the experimental rats were exposed to NaF (lower dose group:5 mg/L,higher dose group:50 rmg/L) added to the drinking water to establish the chronic fluorosis model.All rats were killed on the six months and the metaphysic of femoral was collected.Bone fluorine was detected by ashing-fluorin ion selective electrode method.Bone tissues were stained with hematoxylin-eosin and observed under optical microscope.The content of bone alkaline phosphatase (BALP) in rat serum was detected by enzyme-linked immunosorbent assay(ELISA).The expressions of TGF-β1 and IL-6 mRNA and protein in bone were detected by in situ hybridization (ISH) and immunohistochemistry (IHC).Results The contents of bone fluorine were increased gradually in the control,the lower and higher doses fluoride groups[(306.04 ± 12.57),(652.91 ± 51.83),(1 094.11 ± 91.41)mg/kg,F =31.14,P ＜ 0.05].Bone sclerosis could be observed under optical microscope in lower and higher dose groups.The content of BALP in serum increased with the dose of fluoride gradually in the control,the lower and higher doses fluoride groups[(27.78 ± 4.09),(46.59 ± 5.75),(57.45 ± 3.99)U/L],expressions of mRNA (111.84 ± 4.62,123.86 ± 7.46,140.83 ± 5.21) and protein (118.60 ± 7.09,133.17 ± 7.33,145.67 ± 9.61) of TGF-β1 were both increased(F =30.29,73,28,33.65,all P ＜ 0.05).The expressions of mRNA(117.78 ± 7.01,119.90 ± 5.10) and protein(122.79 ± 6.49,123.81 ± 7.99) of IL-6 were both higher than those of the control (106.49 ± 6.76,112.11 ± 5.80,F =15.47、10.83,all P ＜ 0.05).Conclusion The expressions of osteo-immunology related factor TGF-β1 and IL-6 in bone of rats with chronic fluorosis have changed,which indicates that fluoride can impact the increased bone formation by regulating the micro environment of bone.

Animals ; Apoptosis ; Cell Differentiation ; Cell Proliferation ; Fluoride Poisoning ; metabolism ; pathology ; Fluorosis, Dental ; metabolism ; pathology ; Hedgehog Proteins ; genetics ; metabolism ; Humans ; Osteoblasts ; cytology ; metabolism ; Osteoclasts ; cytology ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology

Objective To study the proliferation and apoptosis and investigate the expression of Indian hedgehog (Ihh),parathyroid hormone-related peptide (PTHrp),smoothened (Smo) protein and mRNA in the cultured rat primary chondrocytes exposed to different doses of NaF.Methods The third generation articular chondrocytes of neonate rat were cultured in vitro and treated with 0 (control),5,10,20 and 40 mg/L of fluoride.The proliferation activities of cells at different times (24,48 and 72 h) were tested by Thiazolyl Blue Tetrazolium Bromide (MTT).The apoptosis rate was determined by flow cytometry.The expressions of protein and mRNA of Ihh,Smo and PTHrp at 48 h were determined by Western blotting and semi-quantitative RT-PCR,respectively.Results After exposed to 5 mg/L of fluoride for 24,48 and 72 h,the proliferation rates were significantly increased [(1.17 ± 0.07)％,(1.20 ±0.06)％,(1.16 ± 0.08)％] compared with those of control group [(1.10 ± 0.08)％,(1.13 ± 0.08)％,(1.15 ± 0.08)％],but the proliferation activity at 48 and 72 h in 40 mg/L group [(0.72 ± 0.11)％,(0.68 ± 0.04)％] was significantly lower than those in control group (all P ＜ 0.05).Compared with the control group,apoptosis rate of cartilage cell in fluoride treatment group increased gradually [(1.47 ± 0.05)％,(19.87 ± 3.03)％,(25.30 ± 1.28)％,(45.73 ± 4.63)％,F =123.328,P ＜ 0.01].Western blot analysis and RT-PCR results showed that the Ihh,PTHrp,Smo mRNA and protein expression increased in the fluoride groups at 48 h (Ihh protein:0.77 ± 0.08 vs.0.98 ±-0.07,1.23 ± 0.06,1.37 ±0.07,1.34 ± 0.07;PTHrp protein:0.68 ± 0.04 vs.0.89 ± 0.05,0.83 ± 0.05,1.29 ± 0.05,1.16 ± 0.08;Smo protein:0.37 ± 0.01 vs.0.64 ± 0.06,0.67 ± 0.03,0.96 ± 0.06,0.69 ± 0.06;Ihh mRNA:0.77 ± 0.05 vs.0.98 ± 0.05,1.09 ±0.05,1.27 ± 0.03,1.46 ± 0.06;PTHrp mRNA:0.67 ± 0.07 vs.0.97 ± 0.05,1.07 ± 0.08,1.37 ± 0.05,1.45 ± 0.05;Smo mRNA:0.45 ± 0.03 vs.0.63 ±-0.04,0.71 ± 0.05,0.81 ± 0.01,1.00 ± 0.02,all P ＜ 0.05).Conclusions Low doses of fluoride can promote the proliferation of chondrocytes cultured in vitro,and high doses of fluoride can promote the apoptosis of chondrocytes cultured in vitro.The expression of Ihh signaling pathway RNAs and proteins of the cartilage cells are increased following increased levels of fluoride.The results suggest that fluorine has activated the Ihh signaling pathway in chondrocytes and promoted the proliferation and apoptosis processes which might be involved in chondrocytes injury.

Objective To investigate the effects of silencing Smo gene on proliferation and apoptosis of rat prima-ry chondrocyte in vitro.Methods The primary chondrocyte was obtained by mechanical-enzyme digestion and identified by Immunohistochemical cells ( ColⅡ) .The animals were divided into control group , control siRNA group and Smo siRNA 1 ~3 group.The siRNA was transfected into chondrocytes by lentivirus vector .After 72 h, the cell viability was detected by MTT, Smo expression was detected by RT-PCR and Western blot, and the apoptosis of chondrocyte was assessed by flow cytometry .Results All types of siRNA were transfected into primary chondrocyte by vectors, the Smo siRNA 1 ~3 may inhibit the expression of Smo mRNA and protein in chondrocytes, and Smo siRNA2 had the highest silencing rate ( the expressions of Smo mRNA and protein were 0.19 ±0.03 and 0.39 ±0.07 ) .The cell viability in Smo siRNA2 group was lowest ( 77.38% ±7.19%) , while the apoptosis rate of Smo siRNA2 was highest ( 21.43%±2.97%) .Conclusions Silencing Smo gene in primary chondrocytes may inhibit proliferation and promote apoptosis , Smo may have a protecting role from apop-tosis of the chondrocyte.

Based on formal concept analysis theory, formal context of formulae, herbs and their relationship was cre-ated in this article first. Then, the created formal context would be divided and optimized according to classification method in Shang Han Lun Lei Fang written by Xu Lingtai, such as Guizhi Tang category, Chaihu Tang category, Xiexin Tang category, and etc. Multilayer complex concept network was used in the description of the universality and particularity of things. The multilayer complex concept network figure of key-formula-classified formulae was completed on the subgroup structure and visualized express of formulae, herbs and their relationship in the Treatise on Exogenous Febrile Disease. This is a new attempt of integrated information technology with diseases harmed by cold in traditional Chinese medicine. It will bring great benefit in relearning traditional classification methods and promoting inheritance and innovation of study on the Treatise on Exogenous Febrile Disease.

AIM:To investigate the effect of piceatannol on the viability, and the abilities of migration and invasion in the prostate cancer cells.METHODS:DU145 cells were treated with piceatannol at different doses (0, 5, 10, 20, 40 and 80 μmol/L) for different time (12, 24, 36 and 48 h) as indicated.The cell viability was assessed by CCK-8 assay.The migration and invasion abilities of the cells were analyzed by wound healing assay and Transwell assay, respectively.The protein levels of p-JAK2 and p-STAT3 were detected by Western blot.RESULTS:Piceatannol dose-dependently decreased the cell viability.After treatment with piceatannol, the abilities of migration and invasion of the cells were significantly inhibited.Moreover, treatment with piceatannol resulted in marked decreases in the protein levels of p-JAK2 and p-STAT3.CONCLUSION:Piceatannol inhibits the viability, migration and invasion of the prostate cancer cells via regulating the JAK2/STAT3 signaling pathway.

Objective:To observe the protein and mRNA expressions of microtubule-associated protein 1 light chain 3 (LC3)B, P62 and Beclin1 in the liver of rats with chronic fluorosis, and to explore the role of autophagy in pathogenesis of liver injury induced by fluorosis.Methods:Using a group design, 54 SD rats were divided into 9 groups according to their weight (100 - 120 g) using a random number table method, each group with 6 rats, half male and half female. They were control group (NC group), low fluoride group (LF group), high fluoride group (HF group), NC + rapamycin (RAP) group, LF + RAP group, HF + RAP group, NC + chloroquine (CQ) group, LF + CQ group, and HF + CQ group. The NC group drank tap water (fluoride concentration was 0.5 mg/L), LF group drank fluoride water (fluoride concentration was 5.0 mg/L), HF group drank fluoride water (fluoride concentration was 50.0 mg/L); NC + RAP group, LF + RAP group and HF + RAP group were fed with corresponding drinking water, respectively, for 3 months, and then RAP (1.5 mg/kg) was intraperitoneally administered for 10 d; NC + CQ group, LF + CQ group and HF + CQ group were fed with corresponding drinking water, respectively, for 3 months, and then CQ (60 mg/kg) was intraperitoneally administered for 10 d. Bone and 24-hour urine samples of rats in each group were collected to detect the contents of bone fluoride and urine fluoride; liver histomorphological changes were observed through hematoxylineosin staining; protein and mRNA expressions of LC3B, P62 and Beclin1 in liver were detected by immunohistochemistry and real-time fluorescence quantitative PCR, respectively.Results:Compared with the NC group [(0.03 ± 0.00) mg/kg, (0.34 ± 0.08) mg/L], the contents of bone fluoride [(3.86 ± 0.08) mg/kg] and urine fluoride [(1.11 ± 0.16) mg/L] in HF group were higher ( P < 0.05). In the NC group, the lobule structure of liver tissue was clear, the hepatic cords were arranged in order, and the cell structure was normal. There were different degrees of hepatocyte edema in LF and HF groups. After intraperitoneal injection of RAP, compared with the corresponding fluoride group, the morphology of hepatocytes did not change significantly. After intraperitoneal injection of CQ, compared with the corresponding fluoride group, the liver cells showed obvious edema, and the degree of edema aggravated with the increase of fluoride concentration. Compared with the NC group, the protein expressions of LC3B and Beclin1 in HF group were higher ( P < 0.05), and the protein expression of P62 was lower ( P < 0.05). After intraperitoneal injection of RAP, the protein expressions of LC3B and P62 in LF + RAP group was lower than that in LF group ( P < 0.05); Compared with HF group, the protein expressions of LC3B and Beclin1 in HF + RAP group were lower ( P < 0.05). After intraperitoneal injection of CQ, protein expression of P62 in LF + CQ group was higher than that in LF group ( P < 0.05); Compared with HF group, protein expression of P62 in HF + CQ group was higher ( P < 0.05). Conclusions:Early (3 month) fluoride intake could promote autophagy and induce edema of hepatocytes in rats, and RAP had similar effects. CQ may induce liver injury by inhibiting autophagy of hepatocytes.

Objective To Investigation the quality of marrow smear purchased by CDUTCM.Methods The quality and the typ-icality of marrow smears purchased during 2015 -2016 were collectively examined,and then decided whether these smears fit the blood cell morphology experimental teaching requirement.Results Of all the 960 marrow smears purchased these two years, 49.7% failed in smear made or stained,and 16.0% failed to meet the teaching requirements in the typicality of marrow cells.Con-clusion Teaching marrow smears,being different from clinic ones in their preparation and morphological diagnosis,must be of great quality in sustaining and of better typicality in their cell features.

This study was aimed to discover the knowledge of urination formula-syndrome in theTreatise on Exogenous Febrile Diseasebased on the Sunshine diagram of multi-layer complex concept network express. A total of 39 items about urination formula-syndrome in theTreatise on Exogenous Febrile Diseasewere collected, and then regulated into standard expression. The database was established and the multi-layer complex concept network express was constructed. The Sunshine diagram was drawn and the connotation rules on urination formula-syndrome in theTreatise on Exogenous Febrile Diseasewere summarized through mode development of the diagram. The results showed that the Sunshine diagram collected 44 objects (i.e., formulas) and 191 properties (i.e. syndromes), which expressed the urination formula-syndrome visually. It was concluded that the application of Sunshine diagram in the formula-syndrome knowledge based on multi-layer complex concept network express provided certain references on the inheritance and development of classics in traditional Chinese medicine (TCM).

Objective To observe the effects of autophagy on the expression of synaptic plasticity related protein, growth-associated pro-tein-43 (GAP-43) and microtubule associated protein-2 (MAP-2), in CA1 area of hippocampus of vascular dementia rats. Methods Nine-ty-six healthy male Sprague-Dawley rats were randomly divided into sham group, vascular dementia model group (VD group), autophagy in-hibitor 3-methyl adenine preconditioning group (3-MA group) and autophagy agonist rapamycin preconditioning group (Rap group). Each group was divided randomly into subgroups of one week, two weeks, four weeks and eight weeks after modeling, six rats in each group. The vascular dementia rat model was established with modified Pulsineli's four-vessel occlusion. The expression of GAP-43 and MAP-2 in CA1 area of hippocampus were detected with immunohistochemistry. Results Compared with the sham group, the expression of GAP-43 protein increased, and the expression of MAP-2 protein decreased at every time point in VD group (P<0.01). Compared with VD group, the expres-sion of both GAP-43 and MAP-2 increased in 3-MA group (P<0.05), and decreased in Rap group (P<0.05). Conclusion Autophagy may in-hibit the expression of synaptic plasticity related protein, GAP-43 and MAP-2, in CA1 area of hippocampus in vascular dementia rats, indi-cating inhibition of autophagy may promote synaptic remodeling.