BACKGROUND:Until now, little has been reported on establishment of pancreatic stem cell strains and lines,and the purification of pancreatic stem cells is difficult since the cell line establish rate is low.OBJECTIVE:To explore a more rational and effective technique of in vitro separation and continuous passage of pancreatic stem cells, with the hope to establish cell strains and even cell lines and to lay the foundation for the follow-up study of pancreatic stem cells in the treatment of diabetes.METHODS:Firstly, Percoll discontinuous density gradient centrifugation method was applied to separate the mouse pancreatic endocrine portion from the exocrine portion, then to obtain cell strains with highly proliferative ability and low differentiation from pancreatic endocrine portion-the islet. We used mouse embryonic fibroblasts treated with mitomycin C as a feeder layer, for in vitro continuous culture of islet-derived pancreatic stem cells under feeder layer conditions until they were transferred to the 30th passage to establish cell lines. Then pancreatic stem cell line derived from pancreatic islet was detected and identified by a series of tests including growth characteristic test, morphological observation, related molecular marker identification and differentiation characteristic identification.RESULTS AND CONCLUSION:In the continuous process of passage, pancreatic stem cells showed active proliferative ability, and maintained the typical morphological characteristics of stem cells and expression of pancreatic stem cell marker-Nestin. After induction, pancreatic stem cells showed insulin gene expression,reflecting their differentiation potential. Therefore, under the condition of feeder layer, the pancreatic stem cell line derived from Kunming mice was successfully established and the related identification was completed,which lays the foundation for the following research.

OBJECTIVETo study the therapeutic effects of Rhizoma Drynariae and estrogen on osteoporosis in ovariectomized (OVX) rats.

METHODSFifty-five female rats were randomly divided into 5 groups, the normal control (normal),the sham operated (sham), the model, the estrogen, and the Rhizoma drynariae (RD) groups; ovariectomized rats were used as postmenopausal osteoporosis model. The changes of morphology and dynamic parameters in different groups were determined by bone histomorphometry.

RESULTSCompared with the model group, the trabecular volume (TBV/TTV) , trabecular thickness (MTPT) and density (MTPD) in the other four groups were significantly increased, while the trabecular template spacing (MTPS) and the ratio of trabecular surface to trabecular volume (TBS/TBV) significantly decreased (P <0. 05); and the osteoid surface (TOS), single label surface [Sfract (s) ] ,double label surface [Sfract (d) ] and bone formation rate (Svf) also decreased,while osteoid maturation period (OMP) increased in the latter four groups. No significant difference of cortical width (MCW) was found between these 5 groups. Compared with the normal and sham groups, TOS, Sfract ( s) , Sfract ( d) , Svf in the estrogen and RD groups increased significantly, while OMP decreased; no significant difference was found in other parameters.

CONCLUSIONRhizoma Drynariae has the similar effect with estrogen in maintaining normal trabecular structure and connection by inhibiting the increased bone turnover of postmenopausal osteoporosis.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Estrogens ; therapeutic use ; Female ; Humans ; Osteoporosis ; drug therapy ; Ovariectomy ; Polypodiaceae ; Postmenopause ; Rats

Pathological cardiac hypertrophy is a maladaptive response in a variety of organic heart disease(OHD),which is characterized by mitochondrial dysfunction that results from disturbed energy metabolism. SIRT3, a mitochondria-localized sirtuin, regulates global mitochondrial lysine acetylation and preserves mitochondrial function. However, the mechanisms by which SIRT3 regulates cardiac hypertrophy remains to be further elucidated. In this study, we firstly demonstrated that expression of SIRT3 was decreased in AngiotensionⅡ(AngⅡ)-treated cardiomyocytes and in hearts of AngⅡ-induced cardiac hypertrophic mice. In addition, SIRT3 overexpression protected myocytes from hypertrophy, whereas SIRT3 silencing exacerbated Ang II-induced cardiomyocyte hypertrophy.In particular,SIRT3-KO mice exhibited significant cardiac hypertrophy. Mechanistically, we identified NMNAT3 (nicotinamide mononucleotide adenylyltransferase 3), the rate-limiting enzyme for mitochondrial NAD biosynthesis, as a new target and binding partner of SIRT3.Specifically,SIRT3 physically interacts with and deacety-lates NMNAT3,thereby enhancing the enzyme activity of NMNAT3 and contributing to SIRT3-mediated anti-hypertrophic effects.Moreover,NMNAT3 regulates the activity of SIRT3 via synthesis of mitochon-dria NAD.Taken together,these findings provide mechanistic insights into the negative regulatory role of SIRT3 in cardiac hypertrophy.Sirtuin 3(SIRT3),a mitochondrial deacetylase that may play an impor-tant role in regulating cardiac function and a potential target for CHF

OBJECTIVETo observe the clinical efficacy of Tiaozhi Jiangtang Tablet (TZJT) on patients suffering from diabetes mellitus with blood stasis syndrome.

METHODSSixty patients were randomly assigned into 2 groups, the TZJT group (n = 30) treated with TZJT tablet and asprin, the control group (n = 30) treated with asprin alone.

RESULTSThe improvement of symptoms was more significant in the TZJT group than that in the control group (P < 0.05). Levels of serum endothelin (ET), nitric oxide (NO) content and blood viscosity were decreased in both groups after treatment, and the effect of TZJT plus asprin was superior to that of asprin alone.

CONCLUSIONTZJT combined with asprin is effective in improving the serum content of ET and NO and reducing blood viscosity.

Adult ; Aged ; Aged, 80 and over ; Coronary Artery Disease ; drug therapy ; etiology ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy

OBJECTIVETo investigate the relationship between Transitional CK19 positive cells and hepatic precancerous lesion in chronic hepatitis B patients.

METHODSWe observed the expression of CK19 in liver tissue of chronic hepatitis B patients by LSAB immunohistochemical staining, and examined serum AFP and ultrasonography one time per 3 months for one year.

RESULTSWe observed a population of CK19 positive cells-with size and structure between those of human oval cells and mature hepatocytes-that usually occurred along with oval-cell proliferation. It was suggested that these transitional cells may partly account for the elevation of serum AFP. One patient occurred hepatic carcinoma, another patient had low-echogenic nodules in liver parenchyma within the 1 year follow-up period.

CONCLUSIONTransitional CK19 positive cells could be regarded as a new possible pathological marker of hepatic precancerous lesion.

Adolescent ; Adult ; Biomarkers, Tumor ; Female ; Humans ; Keratins ; analysis ; Liver Neoplasms ; chemistry ; pathology ; Male ; Middle Aged ; Precancerous Conditions ; chemistry ; pathology ; Tomography, X-Ray Computed ; alpha-Fetoproteins ; analysis

OBJECTIVETo investigate the bonding properties and interface characterization of a domestic 3mol yttrium-stabilized tetragonal zirconium polycrystal (3Y-TZP) framework fired on with 4 different veneering ceramics for zirconia.

METHODS4 different commercial veneering ceramics for zirconia (VITA VM9, SHOFU VINTAGE ZR, IPS e.max Ceram, Cercon ceram kiss) were sintered on 3Y-TZP rectangulars (15 mmx5 mmx5 mm) according to the manufacturers' instructions for shear bond strength test, a metal-ceramic system(Ni-Cr alloy/VITA VMK95) was fabricated in the same type as a control group. Two bilayered specimens (3Y-TZP/VITA VM9, Ni-Cr/VMK95) were prepared for scanning electron microscope (SEM) and energy distribution spectrum (EDS).

RESULTSThe values of shear bond strength test were (18.83 +/- 1.77) MPa for 3Y-TZP/VITA VM9, (23.83 +/- 7.05) MPa for 3Y-TZP/SHOFU VINTAGE ZR, (17.87 +/- 2.30) MPa for 3Y-TZP/IPS e.max Ceram, (22.26 +/- 7.45) MPa for 3Y-TZP/Cercon ceram kiss, (20.55 +/- 5.13) MPa for Ni-Cr alloy/VITA VMK95. There was no statistically significant between all-ceramic groups and the control group (P > 0.05). The failure modes in all-ceramic groups showed predominately adhesive at the interface. SEM showed the 3Y-TZP/VITA VM9 contacted tightly at the interface, while EDS detected Si element diffused into 3Y-TZP material.

CONCLUSIONThe results indicate that domestic 3Y-TZP has a good interface compatibility with 4 commercial veneering ceramics, as a dental framework material, it can satisfy the clinical requirements.

Ceramics ; Dental Bonding ; Dental Porcelain ; Dental Stress Analysis ; Dental Veneers ; Materials Testing ; Yttrium ; Zirconium

OBJECTIVETo select optimal media and conditions for the tissue culture of Dendrobium lituiforum.

METHODThe basic media, light illumination condition, phytohormone, and natural aqueous extracts. were tested and compared. The chlorophyll contents and soluble protein contents were measured.

RESULTN6 medium was suitable for embryo germination and growth. 1/2 MS + NAA 0.5 mg x L(-1) cordd be used as the optimal protocorm multiplying media. Culture 20 days in darkness in advance, and then in light is useful to embryo germinate. 1/2 MS+ NAA 0-0.5 mg x L(-1) is beneficial to protocorm multiplication largely. N6 + 6-BA 2.0 mg x L(-1) + NAA 0.5 mg x L(-1) + 10% potato juice is useful to protocorm differentiation. Phytohormone and potato juice added to the media promoted chlorophyll content and souble protein content. N6 + NAA 0.5 mg x L(-1) + 10% banana juice is the best medium for plantlet rootage and strengthening.

CONCLUSIONConcentration of inorganic salt, nitrogen source and illumination condition are important to embryo germination and growth. The nitrogen source type has effect on the protocorm multiplication. The chlorophyll contents and souble protein contents may be index to indicate the growth condition of plantlets, which can help to select the optimal media.

Chlorophyll ; analysis ; Culture Media ; Dendrobium ; chemistry ; growth & development ; Light ; Plant Growth Regulators ; pharmacology ; Plant Proteins ; analysis ; Plants, Medicinal ; chemistry ; growth & development ; Tissue Culture Techniques

OBJECTIVETo investigate the diagnosis and treatment of malignant fibrous histiocytoma of the retroperitoneum (MFHR).

METHODSThe clinicopathological features, treatment and prognosis of 31 patients with MFHR were retrospectively analyzed.

RESULTSThe mean age of the patients was 52.7 +/- 14.1 years. Male:Female = 22:9. The primary symptoms were abdominal mass and pain. The average diameter of tumor was 15 cm. The histopathologic types of the tumor were inflammatory, storiform-pleomorphic, myxoid and giant cell in 16, 10, 4 and 1 cases. The overall survival rate of 1-, 3- and 5-year was 61.3% +/- 9.8%, 31.6% +/- 11.3% and 21.1% +/- 11.4% with a median survival time of 17.0 +/- 6.3 months. Complete resection of the tumor was the major prognostic factor. Postoperative radiotherapy of 20 - 45 Gy was able to prolong the median survival from 12.1 +/- 11.6 months of surgery alone to 26.4 +/- 22.0 months of surgery plus postoperative radiotherapy though without statistical significance (P = 0.051). Postoperative CHOP chemotherapy was not shown to be beneficial.

CONCLUSIONChemotherapy remains the important method of cure. The survival in patients with MFHR might be improved by complete resection combined with chemotherapy or/and radiotherapy.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; therapeutic use ; Chemotherapy, Adjuvant ; Cyclophosphamide ; administration & dosage ; Doxorubicin ; administration & dosage ; Female ; Follow-Up Studies ; Histiocytoma, Benign Fibrous ; secondary ; surgery ; therapy ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Pancreatectomy ; Postoperative Period ; Prednisone ; administration & dosage ; Prognosis ; Radiotherapy, Adjuvant ; Retroperitoneal Neoplasms ; pathology ; surgery ; therapy ; Retrospective Studies ; Splenectomy ; Survival Rate ; Vincristine ; administration & dosage

Mycoplasma hyopneumoniae (M .hyopneumoniae) and Mycoplasma hyorhinis (M .hyorhinis) infections are common in China .To investigate the prevalence of M .hyorhinis and M .hyopneumoniae in Jiangsu Province of China ,a mo-lecular epidemiological survey was conducted from 399 nasal swab samples of unvaccinated pigs using nested Polymerase Chain Reaction (nested PCR) .Nasal swab samples were collected from Jiangquhai porcine lean (JQHPL) strain pigs and other West-ern breeds .Clinical samples were taken from each pig and divided into different groups based on ages of pigs (7 ,14 ,21 ,28 , and 35 days) .Results indicated that the prevalence of M .hyorhinis was 70 .9% from different herds in Jiangsu Province in China ,while the prevalence of M .hyopneumoniae was 13 .5% .M .hyorhinis infection was more common in pigs for less than 5 weeks of age compared to M .hyopneumoniae infection .Co-infection was also observed in 30 samples (7 .5% ) in which both M .hyorhinis and M .hyopneumoniae were detected .The M .hyorhinis infection increased as the animals grew from 7 to 35 days .The M .hyopneumoniae infection did not change significantly as the pigs grew older .Significant difference of M .hyorhi-nis infection was observed between other Western breeds and JQHPL pigs (P<0 .001) .JQHPL pigs appear to be more sensi-tive to the M .hyorhinis infection as compared to the other Western breeds .However ,there is no obvious relationship between the breed type and M .hyopneumoniae infection (P>0 .05) .

Objective To assess the effects of gonadotropin-releasing hormone analog on glucose and lipid metabolism in girls with idiopathic central precocious puberty(ICPP).Methods A total of 26 girls (aged 6-8 years,breast stage B2) with ICPP were followed up in Department of Endocrinology,Shenzhen Children's Hospital from Jan.2008 to Jun.2011.Those girls received triptorelin therapy for 12 months.Before and the end of the 6th month,the 12th month of the treatment,body mass index(BMI) was calculated,fasting plasma glucose(FPG),fasting plasma insulin(FPI),total cholesterol,triacylglycerols,high density lipoprotein cholesterol,low density lipoprotein cholesterol,apolipoprotein A,apolipoprotein B and estradiol(E2) were measured.Insulin sensitivity was estimated by homeostasis model assessment of insulin resistance(HOMA-IR).Twenty age-matched prepuberty girls were set as controls.Results 1.Before treatment,BMI,FPG,FPI and HOMA-IR in ICPP girls had no significant difference compared with the controls.2.After 6 months treatment of triptrolin,serum E2 concentration in ICPP girls declined from(30.5 ± 9.8) ng/L at the beginning of treatment to (11.2 ± 4.6) ng/L at the end of 6th month (P ＜ 0.01) ; The end of 12 th month of the treatment,FPG and FPI had no significant difference compared with that before of the treatment,but BMI increased from(16.46 ± 1.10) kg/m2 to(18.35 ± 1.30) kg/m2,the difference was significant(P ＜0.05),HOMA-IR increased from 1.24 ±0.30 to 2.08 ±0.40,the difference was significant(P ＜0.05).3.Lipid metabolism parameters remained unchangeable after 12 months of triptrolin treatment.Conclusion Triptorelin may lead to raise of BMI and HOMA-IR in girls with ICPP at 12 months after treatment.