Objective: To investigate the feasibility of inducing canine bone marrow mesenchymal stem cells (BMSCs) to differentiate into epithelium-like cells in vitro and the basic inducing conditions. Methods: BMSCs were obtained from healthy adult canine bone marrow aspirates and were purified by density gradient centrifugation and anchoring culture. BMSCs of second passage were cultured with 4 different media: K-SFM+EGF+BPE, DMEM+EGF, K-SFM+BPE, and DMEM+FBS. The changes in cell morphology were observed daily under phase-contrast microscopy and cell growth curves of every group were drawn by the continual cell counting. The cells were identified by immunocytochemistry staining and flow cytometer through detecting cytokeratin (CK) and epithelial membrane antigen (EMA) on the 7th and 14th day of induction. Results: A few BMSCs in K-SFM+EGF+BPE group became round or oval with processes on the 7th day of induction; more BMSCs became round or oval and their phenotypic markers were CK+ and EMA+ on the 14th day. FACS analysis showed that CK or EMA positive cells were rare in K-SFM+EGF+BPE group on the 7th day after induction (<1%), and the number of positive cells were increased (> 7%) on the 14th day. Cell morphology had no obvious change in DMEM+EGF, K-SFM+BPE, and DMEM+FBS groups during the whole culture process. Moreover, cells in DMEM+EGF, K-SFM+BPE, and DMEM+FBS group were negative for CK and EMA. Conclusion: Our experiment indicates that the combination of K-SFM, EGF and BPE can induce canine BMSCs to differentiate into epithelium-like cells in vitro.

1.0?10~7 copies/mL. The HBsAg IHC staining positive cells could be observed in 6 placental tissues and 3 fetus' liver tissues,and HBcAg was also positive in 1 case of fetus' liver tissue.After co-incubating the tropho- blastic cells and HBV DNA positive serum in vitro,HBsAg expression and HBV DNA could be detected.Apoptosis of HBV-infected trophoblastic cells increased,which was demonstrated by in vivo and in vitro experiments and the apoptosis of placental cells was correlated with the cord blood HBV DNA level.The results of in vitro experiments showed that the apoptosis of trophoblastic cells increased with the elongation of infection time.After 6 months,1 of 12 newborns was positive for HBsAg,HBeAg and anti-HBc,6 was positive for anti-HBs.Conclusions The mechanism of HBV intra-uterine infection may be that HBV breaches the placental barrier and infects the fetus.The localization and replication of HBV in fetal tissues and organs are probably the important factors of chronic HBV infections in neonates.The apoptosis of trophoblastic cells may be the protective mecha- nism for the placental barrier to block the HBV intra-uterine transmission.

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

ObjectiveTo investigate the clinic and pathologic features of one patient diagnosed with neurocutaneous melanosis ( NCM ) by biopsy.MethodsA 21-year-old woman presented with a 2-month history of tinnitus,headache,vomiting and 1-month history of impaired vision.At birth,a massive nevus covering most of the posterior abdomen had been noted as well as the presence of multiple smaller lesions all over the body.Magnetic resonance imaging demonstrated a posterior fossa cyst compatible with the Dandy-Walker syndrome and extensive leptomeningeal enhancement. Surgery was performed to cystectomy and to obtain pathologic specimens from the leptomeninges. Biopsy and immunohistochemical study was performed.ResultsAt surgery,diffuse black pigmentation of the leptomeninges and the cyst was found.Under microscope,the cyst and leptomeninges were composed with melanocytes with variable pigmentation.Those cells positive for HMB45,MelanA,S100 and vimentin.Ki-67 positive cells ＜ 1％.The pathologic diagnosis wasleptomeningeal diffusemelanocytosis. Thepatientdied 2months after thesurgery.ConclusionsNCM is characterized by a focal or diffuse proliferation of melanin-producing cells in both the skin and the leptomeninges.NCM could be compatible with the Dandy-Walker syndrome.Definite diagnosis relies upon the histological data obtained by mean of biopsy.

OBJECTIVETo prepare andrographolide composite particles, and evaluate their particle structure and dissolution.

METHODThe mechanical crushing method was adopted to prepare andrographolide and polyethylene glycol (PEG) 6000 composite particles. The structures were characterized by the scanning electron microscope (SEM) and the differential scanning calorimeter (DSC). The contact angles were determined by the contact angle analyzer. The in vitro dissolution curve was detected.

RESULTAndrographolide and PEG 6000 gave rise to coated composite particle structures, with the decrease in the crystallinity of andrographolide. The in vitro dissolution rate of composite particles was significantly obvious than that of its raw materials, ultrafine powder and their physical mixtures.

CONCLUSIONAndrographolide composite particles based on the mechanical crushing method could notably enhance the in vitro dissolution of andrographolide.

Calorimetry, Differential Scanning ; Chemistry, Pharmaceutical ; methods ; Diterpenes ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Particle Size ; Solubility ; Torsion, Mechanical

BACKGROUND:Borosilicate cannot only be mineralized to form hydroxy carbonate apatite layer, but also have strong chemical reactivity to promote bone cel regeneration. OBJECTIVE:To investigate the effect of the borosilicate bioglass on the growth behavior of rabbit osteoblasts through in vitro culture experiment. METHODS:The initial and secondary extracts of borosilicate bioglass were prepared according to the requirement of ISO10993-12: 2007. The bone marrow mesenchymal stem cels of rabbits were isolated and cultured. The second generation bone marrow mesenchymal stem cels were induced to differentiate into osteoblasts. The osteoblasts of the 5th-15th RESULTS AND CONCLUSION:The osteoblasts proliferation in the initial extract and secondary extract groups was better than that in the α-MEM medium group (P < 0.05). The osteoblasts proliferation in the initial extract group was better than that in the secondary extract group (P < 0.05). The total protein content of osteoblasts in the initial extract group was higher than that in the secondary extract and α-MEM medium group (P < 0.05). There were no significant differences in the alkaline phosphatase activity, apoptosis rate, horizontal migration distance of osteoblast and transmembrane cel number in Transwel between these three groups. These results demonstrate that borosilicate bioglass has good biocompatibility and has a certain benign regulatory role in generations were obtained and cultured with the initial and secondary extracts of borosilicate bioglass and α-MEM medium, respectively. The effects of borosilicate bioglass on the osteoblasts proliferation, protein synthesis, alkaline phosphatase activity, cel apoptosis, and cel migration in horizontal and vertical direction were observed.RESULTS AND CONCLUSION: The osteoblasts proliferation in the initial extract and secondary extract groups was better than that in the α-MEM medium group (P < 0.05). The osteoblasts proliferation in the initial extract group was better than that in the secondary extract group (P < 0.05). The total protein content of osteoblasts in the initial extract group was higher than that in the secondary extract and α-MEM medium group (P < 0.05). There were no significant differences in the alkaline phosphatase activity, apoptosis rate, horizontal migration distance of osteoblast and transmembrane cell number in Transwell between these three groups. These results demonstrate that borosilicate bioglass has good biocompatibility and has a certain benign regulatory role in osteoblast proliferation.

Objective To investigate the clinical efficacy of meloxicam combined with glucosamine sulfate in the treatment of patients with osteoarthritis.Methods Ninety cases of knee osteoarthritis were randomly divided into control group and treatment group,each group 45 cases.Control group was given meloxicam tablets 7.5 mg,bid,oral.On the basis of control group,treatment group was treated with glucosamine sulfate capsules 0.5 g,tid,oral.All the patients were treated for 6 weeks.The clinical symptoms,signs,knee joint motor function and pain indexes of all patients were evaluated before and after treatment.The blood samples and joint fluid specimens were collected,to determine the matrix metalloproteinase-3 (MMP-3),matrix metalloproteinase-13 (MMP-13) and prostaglandin E2 (PGE2) levels by enzyme linked immunosorbent assay (ELISA).The clinical efficacy and adverse drug reactions in two groups were compared.Results After treatment,the levels of Lequesne indexes in treatment group and control group were 5.78 ± 3..04 and 8.24 ± 3.62,Lysholm scores were 85.82 ± 10.01 and 72.46 ± 13.58,VAS scores were 2.45 ± 1.08 and 3.86 ± 1.17,serum MMP-3 levels were (114.69 ± 13.54) and (130.52 ± 16.48) ng · L-1,serum MMP-13 levels were (141.06 ± 21.55) and (179.82 ± 24.37) ng · L-1,serum PGE2 levels were(69.82 ± 18.67)and(91.30 ±20.09)ng · L-1,joint fluid MMP-3 levels were(17.89 ±5.12) and (23.10 ±5.46)ng · L-1,joint fluid MMP-13 levels were(27.35 ±6.52)and(38.47 ±6.85)ng · L-1,joint fluid PGE2 levels were (82.16 ± 22.64) and (118.02 ± 23.41) ng · L-1,all with significant difference (all P ＜ 0.05).The improvement rates of control group and treatment group were 73.33％ (33 cases/45 cases),91.11％ (41 cases/45 case),with significant difference (P ＜ 0.05).No adverse drug reactions occurred during the treatment.Conclusion The clinical effect of meloxicam combined with glucosamine sulfate in the treatment of knee osteoarthritis is exact,the treatment is conducive to the improvement of clinical symptoms,can reduce serum MMP-3,MMP-13,PGE2 levels,with high safety.

This study was aimed to investigate the effects of DMSO on platelets during pre-treatment for lyophilization, including centrifugation, washing and loading trehalose. After pre-treatment for lyophilization, the expression of platelet membrane surface glycoprotein (GP) including CD62p and PAC-1 was analyzed by FCM before and after induction with thrombin, the mean platelet volume (MPV) and platelet maximal aggregation with several platelet inducers were investigated. The results showed that the expression rates of CD62p and PAC-1, as the platelet activation signs, increased and were 30.37% and 15.01% respectively in group without DMSO after pre-treatment. And their differences in comparison with control were statistically significant, but that of CD62p was 10.72% and PAC was 10.11% in group with DMSO, in comparison with group without DMSO respectively, their differences were statistically significant after diluting with DMSO, CD62p was re-expressed to 54.39% in group with DMSO and more than that in group without DMSO and lower than control statistically significant. PAC-1 was re-expressed to 49.28% in group with DMSO and more than that in group without DMSO (p<0.01) and reached to control. Platelet maximal aggregations induced by thrombin, restocetin and propyl gallate were 92.76%, 91.24% and 89.66 respectively in group with DMSO. These were closed to that in control group and in group without DMSO. But the aggregation induced by ADP was 34.33%, it was less than control (p<0.01) and more than that in group without DMSO (p<0.01). It is concluded that DMSO can inhibit the expression of CD62p and PAC-1 on platelet in vitro. But when diluted with plasma, platelets can express CD62p and PAC-1 induced by thrombin and be led to aggregate by several inducers, so the inhibitory effects of DMSO on platelet activation are reversible. DMSO play roles in inhibitor damage from platelet activation and cryoprotectant. This property of DMSO is very important in research of platelets lyophilization.
Blood Platelets ; cytology ; drug effects ; metabolism ; Blood Preservation ; methods ; Cell Survival ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Dimethyl Sulfoxide ; pharmacology ; Freeze Drying ; Humans ; Platelet Activation ; drug effects ; physiology ; Trehalose ; blood ; pharmacology

OBJECTIVETo provide scientific evidence of making measures for prevention of pesticide poisoning, the investigation on the condition of pesticides poisoning was carried out in Wuhan.

METHODSRegistration data of pesticide poisoning from 2002 to 2010 in Wuhan were collected and statistically analyzed by SAS 9.1.

RESULTSDuring the nine years, there were 4713 cases reported for pesticide poisoning. Among them, the number of occupational poisoning was 2737 (2 cases died), with fatality rate of 0.07%. The number of non-occupational poisoning was 1976 (159 cases died), and its fatality rate was 8.05%. The incidence of occupational poisoning and non-occupational poisoning accounted for 58.1% and 41.9%, respectively. Insecticides especially organophosphorus insecticides, such as parathion, dichlorvos, and methamidophos accounted for 70.6% of the poisoning. Occupational poisoning took place mainly in man, accounting for 68.8%, Non-occupational or life poisoning in contrast mainly occurred in women with a proportion of 66.8%. The majority of the occupational poisoning were 30-59 year-old patients (2239 cases, 81.8%). The majority of the non-occupational poisoning were 30-44 year-old patients (665 cases, 33.6%) and - 70 years old patients (209 cases, 10.6%). High incidence of occupational pesticide poisoning, the regional distribution of Caidian (1016 cases, 37.1% ) highest, followed by the Dongxihu, Hannan and Huangpi. The pesticide poisoning mainly occurred from July to September. The occupational poisoning was mainly caused by poor protection, long working hours, and practice not implemented. The non-occupational poisoning was mainly caused by suicide.

CONCLUSIONSThe majority of the occupational poisoning in Wuhan was middle-aged men. The pesticide poisoning was main caused by insecticides.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Disease Notification ; Female ; Humans ; Infant ; Male ; Middle Aged ; Pesticides ; poisoning ; Young Adult

Tripterygium wilfordii Hook has the properties of rheumatism dispelling,promoting blood circulation, dredging collaterals, detumescence, pain relieving, anti-parasitism and detoxification. Its common medication forms include Tripterygium wilfordii Hook tablets, Tripterygium wilfordii Hook polygonal tablets, Kunming Shanhaitang tablets,Kunxian Capsules,Tripterygium wilfordii Hook bicolor and Tripterygium wilfordii Hook total terpene tablets. Reproductive toxicity in Tripterygium wilfordii Hook preparations is a side-effect that may deeply impact reproductive-age patients. The combination of single herb or compound can significantly reduce the reproductive toxicity of Tripterygium wilfordii Hook preparations.This article reviewed the recent research in this field and provide references for clinical medication.