Objective To study osteogenic courses and features of human osteoblasts cultured in vitro and seeded into alloplastic decalcified cancellous bone.Methods Human osteoblasts isolated from normal human perios- teum were cultured and amplified in vitro.Thy configuration and developing manners of osteoblasts were observed. Then osteoblasts were seeded into alloplastic decalcified cancellous bone.Thy composites were implanted and cultured in thy body of athymic mice.The specimens were obtained 8 weeks later.Osteogenic characteristics were observed by scanning electron microscope.Results Osteoblasts isolated from human periosteum developed well and proliferated rapidly in vitro.Island new bone formation could be observed histologically 8 weeks after the composites of os- teoblasts and alloplasric decalcified cancellous bone were implanted and cultured in the body of athymic mice.Conclu- sion Calcified bone can be formed by seeding osteoblasts into alloplastic decalcified cancellous bone,which may be a new kind of bone graft source to construct bone defects.

In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

OBJECTIVETo observe the effects of cucurmosin (CUS) on the cell proliferation and apoptosis in pancreatic PANC-1 cells.

METHODSThe inhibition of CUS on the PANC-1 cell growth was observed using MTT assay. The inhibition ratio of CUS on the pancreatic orthotopic transplantation was in vivo observed in the NOD/SCID mouse model. The changes of microstructure of the apoptosis-inducing effect of CUS on PANC-1 was observed under electron microscope. The cell cycle and apoptosis after CUS intervention was detected using flow cytometry. The Caspase-3 activity after CUS treatment was detected using enzyme linked immunospecific assay (ELISA).

RESULTSTreatment with CUS at the dose of 0.125, 0.25, and 0.5 mg/kg inhibited the growth of pancreatic carcinoma PANC-1 xenografs with the ratio of 45.2%, 50.0%, and 59.7%, respectively (P < 0.05). After exposure to 10 microg/mL CUS for 24 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation and shrunken nucleus. The apoptotic cells increased. Some nuclear shrinkage and fragmentation, as well as the apoptotic body were observed when cells were exposed to CUS for 72 h. Being exposed to 0, 2.5, 10.0, and 40.0 microg/mL of the CUS for 72 h, the percentage of G0/G1 phase cells was 46.56% +/- 5.08%, 53.33% +/- 5.05%, 67.50% +/- 6.50%, and 77.00% +/- 6.73%, respectively (P < 0.05). The apoptosis ratio was 2.50% +/- 0.13%, 8.30% +/- 1.23%, 23.40% +/- 2.45%, and 48.50% +/- 3.65% shown by Annexin V/PI (P < 0.05). The Caspase-3 activity (unit) was 0.009 +/- 0.002, 0.011 +/- 0.003, 0.035 +/- 0.009, and 0.065 +/- 0.009, respectively (P < 0.05). These data showed that CUS induced the apoptosis of PANC-1 cells in a dose-dependent maner. Being exposed to 40.0 microg/mL of the CUS for 24, 48, and 72 h, the percentage of G0/ G1 phase cells was 56.60% +/- 6.65%, 67.83% +/- 6.76%, and 77.00% +/- 6.73%, respectively (P < 0.05), the apoptosis ratio was 16.51% +/- 2.97%, 38.51% +/- 2.38%, and 48.50% +/- 3.65% shown by Annexin V/PI (P < 0.05). These data showed that CUS induced apoptosis of PANC-1 cells in the G0/G1 phase of the cell cycle in a time-dependent maner.

CONCLUSIONCUS significantly inhibited the growth of PANC-1 cells possibly through the G0/G1 cell cycle arrest and apoptosis.

Animals ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cucurbita ; Female ; Humans ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Pancreatic Neoplasms ; pathology ; Plant Proteins ; pharmacology

OBJECTIVETo investigate the clinical effects of the Chinese traditional medicine Yougui Capsules and Wuziyanzong Pills on sperm viability and motility in patients with oligoasthenospermia.

METHODSA total of 80 infertile men oligoasthenospermia were equally randomized into a trial and a control group, the former treated with Yougui Capsules at 1.68 g tid, while the latter with Wuziyanzong Pills at 6 g bid, both for a course of 12 weeks. The sperm viability and motility of the patients were detected and compared before and after medication.

RESULTSAfter 12 weeks of medication, the sperm viability and percentages of grade a and grade a + b sperm were (65.7 +/- 13.1), (22.5 +/- 9.1) and (47.6 +/- 15.8)% in the trial group, significantly higher than (38.1 +/- 11.1), (13.2 +/- 6.8) and (24.1 +/- 10.9)% in the control (P<0.05). What's more, the above three parameters of the two groups were also significantly higher than those before medication, which were (31.9 +/- 16.9), (8.2% +/- 3.7) and (15.7 +/- 13.9)% in the former and (31.7 +/- 17.0), (7.9 +/- 4.5) and (16.9 +/- 13.6)% in the latter (P<0.05).

CONCLUSIONBoth Yougui Capsules and Wuziyanzong Pills can improve sperm viability and motility in patients with oligoasthenospermia, and the former is even more efficacious than the latter.

Adult ; Asthenozoospermia ; therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Young Adult

OBJECTIVETo study the expression of S phase kinase associated protein 2 (Skp2), phosphatase and tensin homolog deleted on chromosome ten (PTEN) in human laryngeal carcinoma and precancerous lesions, to explore their relations and clinical significance.

METHODSFormalin-fixed and paraffin-embedded tissues from 79 cases of laryngeal carcinoma, 16 cases of atypical hyperplasia of vocal fold,14 cases of adult laryngeal papillomas and 27 cases of vocal cord polyps were evaluated for the expression of Skp2, PTEN by SP immunohistochemistry, the levels of these proteins in tissues with the different types of lesion and their correlation with clinicopathological parameters of laryngeal carcinoma were analyzed.

RESULTSThe expression rates of Skp2 in vocal cord polyps, adult laryngeal papillomas, atypical hyperplasia of vocal cord and laryngeal carcinoma were 11. 11% ,14. 29% ,37. 50% ,39. 24% respectively. There was significant difference among them( Hc = 11. 57, P <0. 01). The expression rates of PTEN protein in vocal cord polyps,adult laryngeal papillomas, atypical hyperplasia of vocal cord and laryngeal carcinoma were 100% ,92. 86% ,75. 00%, 56. 96% respectively . There was significant difference among them (Hc = 62. 86, P<0. 05). There was a negative correlation between the expression of Skp2 and PTEN,and their correlation coefficient was r = -0. 4512(P <0. 01). Patients with Skp2 expression in laryngeal carcinoma revealed poorer five yeas survival rate than patients with negative expression of Skp2 (x2 = 21. 46, P = 0. 000).

CONCLUSIONSThe expression of Skp2 and PTEN took important roles in the tumorigenesis, aggressiveness, metastases of laryngeal carcinoma. The high expression of Skp2 was negative correlation with the lower PTEN in laryngeal carcinoma, which suggested that PTEN may regulate the expression of Skp2.

Adult ; Aged ; Aged, 80 and over ; Carcinoma ; pathology ; Female ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; PTEN Phosphohydrolase ; metabolism ; S-Phase Kinase-Associated Proteins ; metabolism

OBJECTIVETo investigate the effects of nitric oxide (NO) on reperfusion injury following pancreaticoduodenal transplantation in rats.

METHODSThe homologous male Wistar rat model of heterotopic total pancreaticoduodenal transplantation was used. The L-arginine (L-Arg) group received intravenous injection of L-Arg 5 minutes before and after reperfusion at a dose of 200 mg/kg while the N-Nitro-L-Arginine methyl ester (L-NAME) group received intravenous injection of L-NAME at a dose of 10 mg/kg, and control group received saline. The amount of NO in the pancreas graft was measured. Serum concentration of cytokine-induced neutrophil chemoattractant (CINC) determined by enzyme-linked immunosorbant assay, expression of CINC mRNA detected by Northern blot assay, and myeloperoxidase (MPO) activity in the pancreas graft were measured. Histological observation was performed.

RESULTSThe amount of NO in the L-Arg group was higher than in the control group, while in the L-NAME group was lower than in the control group (P < 0.05). The peak of serum CINC concentration occurred 3 hours after reperfusion with significant difference among groups. Expression peak of CINC mRNA in the pancreas graft occurred 3 hours after reperfusion. The expression level in the L-Arg group was lower than in the control group, the L-NAME group was higher than control group (P < 0.05). MPO activity in the L-Arg group obviously decreasd compared with other groups. The pancreas inflammation was ameliorated in L-Arg group, and pancreas damage was aggravated in L-NAME group.

CONCLUSIONSL-Arg can increase the amount of NO and inhibit the elevation of CINC, CINC mRNA expression, and early neutrophil accumulation in the transplanted pancreas. NO has protective effects on the ischemia/reperfusion injury of pancreaticoduodenal transplantation.

Animals ; Arginine ; pharmacology ; Chemokines, CXC ; biosynthesis ; genetics ; Duodenum ; transplantation ; Male ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; metabolism ; Pancreas Transplantation ; Peroxidase ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism

OBJECTIVETo investigate the protective effect and mechanism of Shenqi Compound on diabetic angiopathy modeled rats.

METHODSTotally 18 SD rats were randomized into 3 groups, i.e., the normal control group, the diabetic mellitus (DM) group, and Shenqi Compound group, 6 in each group. The DM rat model was established by feeding high-fat diet (to induce hyperlipidemia) +intraperitoneal injection of small dose streptozotocin (STZ). Shenqi Compound was given to rats in the Shenqi Compound group at the daily dose of 2 g/kg. Equal volume of normal saline was given to rats in the model group and the normal control group by gastrogavage. All treatment was lasted for 12 weeks. Then 2-D and ultrasonic integrated backscatter technique were used to evaluate structural and functional changes of abdominal aorta in the progression of diabetic macroangiopathy. The fibrosis degree of the aorta vessel and myocardium capillaries were observed by using HE and Masson trichrome staining. The tension of the aortic vascular ring was determined. The transforming growth factor beta (TGF-beta) mRNA expression was detected by real time PCR (RT-PCR). The protein expression of TGF-beta, collagen I, collagen III, connective tissue growth factor (CTGF), and phosphorylation P38 MAPK were detected by Western blot.

RESULTSCompared with the normal control group, abdominal aortic systolic inner diameter, diastolic inner diameter, Peterson elastic modulus, stiffness index, and backscatter integral significantly increased; the rangeability of integral backscatter and the extension coefficient of cross section significantly decreased in the DM group (all P < 0.05). After 12 weeks aforesaid indices were obviously improved in the Shenqi Compound group (P < 0.05). Results of HE and Masson staining showed that the fibrosis degree of the aorta vessel and myocardium capillaries was obviously alleviated in rats of the Shenqi Compound group (P < 0.05). Results of the aortic vascular ring tension showed that acetylcholine induced vasodilatation and maximum diastolic percent were obviously elevated in the Shenqi Compound group (P < 0.05). Compared with the normal control group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all significantly increased in the DM group (P < 0.05). Compared with the DM group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all decreased (P < 0.05).

CONCLUSIONSShenqi Compound could effectively improve the arterial function in diabetic marcoangiopathy and microvascular dysfunction. The mechanism might be due to the down-regulating the expression of TGF-beta, and further suppressing the phosphorylation of P38 MAPK, reducing the synthesis of collagen I and collagen III, therefore, ameliorating arterial and myocardial interstitial fibrosis.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Angiopathies ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the expression of cyclooxygenase-2 (COX-2) in different prostate cancer (PCa) cell lines and its role in the acquisition of invasive and metastatic potentials of PCa.

METHODSWe detected the expressions of COX-2 in prostate cancer cell lines LNCaP, C4-2, IF11, IA8 and PC-3 with different metastatic potentials by Western blotting and RT-PCR, and analyzed their roles in the invasion and metastasis of different PCa cell lines.

RESULTSWestern blotting and RT-PCR showed that the expression of the COX-2 protein was high in PC-3, but absent in IF11, IA8, LNCaP and C4-2 (P < 0.05), and it was consistent with the expression of COX-2 mRNA.

CONCLUSIONCOX-2 expresses differently in PCa cell lines with different metastatic potentials. The overexpression of COX-2 may be associated with the high invasion and metastasis of PC-3, but not with the metastasis of other cell lines.

Cell Line, Tumor ; Cyclooxygenase 2 ; metabolism ; Humans ; Male ; Neoplasm Metastasis ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics

OBJECTIVETo explore the effective therapeutic method in the treatment of idiopathic sudden hearing loss (ISHL).

METHODSOne hundred and eighty-eight cases of ISHL were randomized into a warming-promoting needling group (74 cases), a conventional acupuncture group (56 cases) and a medication group (58 cases). In the conventional acupuncture group, the conventional needling technique was applied to Baihui (GV 20), Fengchi (GB 20), Yifeng (TE 17), Tinggong (SI 19), Touqiaoyin (GB 11) and Zhigou (TE 6) on the affected side. The treatment was given 5 times each week. Totally, the treatment of 6 weeks was required. In the warming-promoting needling group, on the basic treatment as the conventional acupuncture group, the warming-promoting needling technique was applied to Fengchi (GB 20). In the medication group, the intravenous drop with salvia injectio and mecobalamin was prescribed, once per day, for 10 days totally. Meanwhile, Erlong Zuoci Wan was prescribed for oral administration, 8 pills each time, three times a day for 30 days continuously.

RESULTSAll of the three therapeutic methods achieved the effect on ISHL. The total effective rate was 89.2% (66/74) in the warming-promoting needling group, which was better than 62.5% (35/56) in the conventional acupuncture group and 53.4% (31/58) in the medication group (both P<0.01).

CONCLUSIONThe warming-promoting needling techinque achieves the significant efficacy on ISHL. The hearing improvement is superior to that treated with either the conventional needling technique or medication.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adult ; Female ; Hearing Loss, Sudden ; therapy ; Humans ; Male ; Middle Aged ; Needles ; Treatment Outcome

To research the effect of Ginseng Radix et Rhizoma and Aconiti Lateralis Radix Praeparata compatibility on cardiac toxicity in rats by UPLC-Q-TOF/MS, and explore the endogenous markers and molecule mechanism. Different compatibility of Shenfu decoction were given to male Wistar rats at dosage of 20 g · kg(-1) for 7 days, collected the serum, and analyze the endogenous metabolites effected by Shenfu formulation by principal component analysis and partial least-squares analysis. Results showed that content of glutathione, phosphatidylcholine and citric acid decreased in mixed-decoction group, while ascorbic acid, uric acid, D-galactose, tryptophan, L-phenylalanine increased. The results showed cardiac toxicity of Aconiti Lateralis Radix Praeparata in Shenfu mixed-decoction. Shenfu co-decoction group showed a similar or weaker trend compared with control group, but most of them do not have a statistically significant. The results indicated the scientific basis of Shenfu compatibility by comparison of co-decoction group with mixed-decoction group. Shenfu compatibility can reduce cardiac toxicity induced by Aconiti Lateralis Radix Praeparata, and citric acid, glutathione, phosphatidyl choline, uric acid might be regarded as potential markers of cardiotoxicity.
Animals ; Biomarkers ; Cardiotoxicity ; Drugs, Chinese Herbal ; toxicity ; Glutathione ; blood ; Least-Squares Analysis ; Male ; Metabolomics ; methods ; Principal Component Analysis ; Rats ; Rats, Wistar