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 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

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.

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.

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

OBJECTIVETo evaluate the effect of branch chain amino acid (BCAA) enriched formula on nutritional status and postoperative fatigue for digestive surgery patients.

METHODSForty patients who underwent digestive surgery were randomly received parenteral nutrition with either BCAA enriched (study group, n=20) or routine amino acid (control group, n=20) for seven consecutive days. Nitrogen balance,serum total protein, albumin, prealbumin, transferrin, retinol binding protein and postoperative fatigue score were monitored during the postoperative period.

RESULTSThe cumulative postoperative fatigue scores were lower in BCAA group than that in the control group at the 4th, 5th and 7th day after operation (P< 0.05). Patients achieved positive nitrogen balance 2 days earlier in the study group than that in the control group, but there was no significant difference in cumulative nitrogen balance between the two groups. There was no significant difference in elevation of serum total protein, albumin, prealbumin, transferrin at the 7th day after operation between the two groups (P > 0.05), compared with those at the first day after operation. The serum level of retinol binding protein was higher in BCAA-enriched group than that in the control group (P=0.004).

CONCLUSIONTPN with BCAA enriched formula can reduce postoperative fatigue score and improve the nutritional status for digestive surgery patients.

Adult ; Aged ; Amino Acids, Branched-Chain ; therapeutic use ; Digestive System Surgical Procedures ; adverse effects ; Fatigue ; etiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Nutritional Status ; Parenteral Nutrition ; Postoperative Period

OBJECTIVEAccording to the information obtained from the revision of total hip arthroplasty, the situations of the artificial femoral stem under physiological stress were analyzed preliminarily. In order to accumulate clinical experience, the discussions about how to enlongate the life of artifical joints were performed.

METHODSFifty-three patients required revision operations were selected, including 28 males and 25 females,with an average age of 74.66 years old (61 to 84 years old). The average life of artificial joints was 18.24 years (3 to 27 years). The initial weightbearing was started 11 days (5 to 16 days) after the first operation. The main reasons for the revision were the spontaneous loosening and trauma. Based on imaging and surgical findings, as well as the histological pathology, the situations of the stems in human bodies were determined.

RESULTSFemoral prosthesis would sink under physiological stress. The design of femoral stems, motion of patients', and bone growth determined the secondary stability. Straight stems were helpful for physiological subsidence of prosthesis, achieving the stability with close bone-implant contact by bone growth,which allowed the patient to do early landing exercise. The collared prosthesis and prosthesis combined with internal fixation limited the subsidence of femoral stems. Their stability depends on the time of initial placement and the bone growth for supporting. Delaying the inital weightbearing was suggested for patients with these protheses.

CONCLUSIONAccording to the actual situation of the patients, the appropriate chosen of femoral stems and time to perform the weightbearing can extend the life of the femoral prosthesis.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; methods ; Female ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Prosthesis Design

OBJECTIVETo investigate the value of acoustic radiation force impulse (ARFI) in evaluating the stage of hepatic fibrosis and early stage cirrhosis.

METHODSSixty-six patients with viral hepatitis underwent liver biopsy and 33 normal subjects (S0) were selected to accept ARFI,the shear wave velocity of hepatic segments s5, s6, s7, s8 and size of liver were measured. The results of liver and spleen size and portal vein's diameter were also measured.

RESULTSThe 66 patients were divided into 3 groups: S1, S2-S3, S4. ARFI for 66 patients and 33 normal subjects showed good image quality. There were statistically significant differences between S4 group and S0 group, S1 group, S2-S3 group for the shear wave velocity of hepatic segments s5, s6, s7, s8 (P < 0.05). Between S2-S3 group and SO group S1 group, the shear wave velocity of hepatic segments s5, s6, s7, s8 also have statistically significant differences (P < 0.05), other parameters showed no significant difference (P > 0.05). Spleen size and the portal vein's diameter of S4 group were larger than those in other groups (P < 0.05).

CONCLUSIONSThe invasive acoustic radiation force impulse could evaluate the stage of hepatic fibrosis and early stage cirrhosis in patients suffering from viral hepatitis. The measurement was feasible. It was suitable for clinical use.

Acoustics ; Adult ; Female ; Humans ; Liver ; pathology ; Liver Cirrhosis ; pathology ; Male ; Middle Aged ; Portal Vein ; pathology ; Spleen ; pathology