AIM: To investigate the action of testosterone on atherosclerosis in a rabbit model. METHODS: 37 male cholesterol-fed rabbits were divided into five groups: castration group: castrated rabbits without exogenous testosterone administration; testosterone Ⅰ group: castrated rabbits with exogenous testosterone 0.25 mg?kg -1?d -1; testosterone Ⅱ group: castrated rabbits with exogenous testosterone 2.5 mg?kg -1?d -1; testosterone Ⅲ group: castrated rabbits with exogenous testosterone 12.5 mg?kg -1?d -1. The sham operation group was also set. Three months later, the levels of testosterone, blood lipids (including TG, HDL-C, LDL-C), PAI activity, nitric oxide (NO) content, endothelin (ET), angiotensin Ⅱ (AngⅡ) in blood were detected. RESULTS: It showed that testosterone in castration group was the lowest. There was no significant difference of TG or LDL-C between castration group and the other four groups. HDL-C in castration group was lower than that in other four groups. NO content of castration group was lower than that in others, but PAI activity, ET and AngⅡ concentration were higher than that in other groups. CONCLUSION: Testosterone is a protective factor against atherosclerosis in male rabbits.

Tetradrine-tashionone II(A)-PLGA composite microspheres were prepared by the SPG membrane emulsification method, and the characterization of tetradrine-tashionone II(A) -PLGA composite microspheres were studied in this experiment. The results of IR, DSC and XRD showed that teradrine and tashionone II(A) in composite microspheres were highly dispersed in the PLGA with amorphous form. The results of tetradrine-tashionone II(A) -PLGA composite microspheres in vitro release experiment showed that the cumulative release amounts of tetradrine and tashionone II(A) were 6.44% and 3.60% in 24 h, and the cumulative release amounts of tetradrine and tashionone II(A) were 89.02% and 21.24% in 17 d. The process of drug in vitro release accorded with the model of Riger-Peppas. Tetradrine-tashionone II(A) -PLGA composite microspheres had slow-release effect, and it could significantly reduce the burst release, prolong the therapeutic time, decrease the dosage of drugs and provide a new idea and method to prepare traditional Chinese medicine compound.
Benzofurans ; chemistry ; Benzylisoquinolines ; chemistry ; Drug Carriers ; chemistry ; Drug Compounding ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Kinetics ; Lactic Acid ; chemistry ; Microspheres ; Particle Size ; Polyglycolic Acid ; chemistry

Objective To investigate the effects of estrogen on mismatch repiar gene expression in colonic mucosa in vivo. Methods A total of 42 healthy individuals underwent colonoscopy were enrolled in the study. Half an hour before colonoscopy examination, blood sample was taken for determining the serum estradiol (E2) level. N ormal colonic mucosal tissues determined by naked eye under colonoscopy examination were taken in the right hemi colon to detect HMLH1 and hMSH2 gene expression by semi-quantitative RT-PCR and immunohistochemistry staining. Then the correlation of serum E2 levels with hMLH1 and hMSH2 expression in colonic mucosa was analyzed. Results A bimodal curve was presented for the correlation between serum E2 level in healthy individuals and hMLH1 expression in colonic mucosa. A strong positive correlation of E2 level with hMLH1 expression in normal colonic mucosa was observed when serum E2 level was more than 45 pg/ml (For mRNA, P=0. 003, r=0. 701; for immunohistochemistry positivity index, P=0. 000, r=0. 874).However there was no correlation between E2 level and hMSH2 expression. Conclusion High serum E2 level might increase the hMLH1 gene expression in colonic mucosa in vivo.

Objective To summarize the experience in diagnosis and therapy of familial adenomatous polyposis with desmoid tumour. Methods Clinical data of 6 patients with familial adenomatous polyposis and desmoid tumour from Jan 1989 to Jan 2005 were retrospectively analyzed. Results Five patients received proctocolectomy and 1 abdominoperineal resection. The most common symptom was progressive painless mass in abdomen. All patients were confirmed by image examination. Four received surgery, 2 were treated by medicine postoperatively and 1 got watchful therapy. Postoperative recurrence developed in two cases and one suffered from short bowel syndrome. All patients were alive at the follow-up. Conclusions FAP with desmoid tumour is not a rare condition and we should pay attention to diagnosis and manage this disease entity. A reasonable remedy was selected according to general state of health and location of desmoid tumour which can improve prognosis and quality of life.

The purpose of this study was to investigate how Zuogui pills from the Kidney-tonifying and Nourishing Yin formula, in combination with the gonadotrophin-releasing hormone antagonist cetrorelix, affected the ovarian local oxidative stress response in decreasing ovarian reserve (DOR) mice. All animal experiments were carried out in accordance with the guidelines and standards established by Jinan University's Experimental Animal Management Committee. Cyclophosphamide (CTX)-treated DOR mice were given Zuogui pills, cetrorelix, or a combination of the two drugs intragastrically. After treatment, there were changes in the estrous cycle, serum sex hormone levels, oxidative stress-related indexes, growth biochemical factor levels, and SIRT1/P53/P21 expression. In comparison to the model group, the Zuogui pills and the cetrorelix+Zuogui pills group had significantly prolonged estrous periods and shortened interestrous periods, and the cetrorelix+Zuogui pills group had a significantly shortened cycle length. Follicle-stimulating hormone (FSH) decreased and estradiol (E₂) increased in all treatment groups compared to the model group, oxidative stress indexes nitric oxide synthase (NOS), nitric oxide (NO), and reactive oxygen species (ROS) decreased, growth biochemical factors brain derived neurotrophic factor (BDNF) and growth differentiation factor 9 (GDF-9) concentrations increased significantly, and leukemia inhibitory factor (LIF) showed no significant change. SIRT1/P53/P21 immunohistochemical results revealed that, when compared to the model group, the expression of SIRT1 increased while the expression of P53 and P21 proteins decreased in all treatment groups, with the cetrorelix+Zuogui pills group having the largest decrease, with significant differences in all indicators. We conclude that cetrorelix combined with Zuogui pills for kidney nourishing and Yin recipe improved the oxidative stress response in the follicle by regulating the SIRT1/P53/P21 pathway, reducing peroxide product production, protecting ovarian function, and regulating ovarian hormone secretion, and its efficacy is superior to that of cetrorelix or Zuogui pills alone.

Seeds of Bupleurum chinense cultivar, Zhongchai No. 1, were sowed in plastic pots which used the arable layer soil as the nursery bed and putted in the artificial climate incubator at various temperatures (15, 20, 25, 15-25 degrees C) and light (8,12 h) to germinate, respectively. The lower constant temperature (15 degrees ) and the higher constant temperature (25 "C) were not conducive to the sprouting characteristics of B. chinese. While they were able to enhance root activity to some extent; The seeding growth of B. chinese was significantly better in the variable temperature than correspondence in the constant temperature, significantly. The emergence speed, emergence index, vigor index and root activity of Bupleurum were improved under the 12 h of light-time, but the germination rate was not improved. The sprouting of Bupleurum's seeds could be improved to some extent by soaking with hormone, such as gibberellin, cytokinin, salicylic acid. Gibberellin promoted seeds' sprouting and seedings's root activity of Bupleurum, while salicylic acid increased the root activity of seeding. There is a significant influence of light, temperatures and hormone treatment on the germination of Zhongchai No. 1 seeds, and all three are remarkably interacted; It is beneficial to promote seed germination by the temperature (20 + 5) degrees C, lighting (8 h) and gibberellin concentration (10 x 10(-6)).
Bupleurum ; drug effects ; growth & development ; radiation effects ; Germination ; drug effects ; radiation effects ; Gibberellins ; pharmacology ; Light ; Plant Growth Regulators ; pharmacology ; Seeds ; drug effects ; growth & development ; radiation effects ; Temperature

BACKGROUND: It has been believed mesenchymal stem cells (MSCs) play a role in treatment through paracrine mechanism. Various side effects such as embolism, tumorigenesis and immunological reaction caused by direct injection of MSCs can be avoided by extracting MSC lysate. However, there is a larger difference in current collection methods and standards of MSC lysate. OBJECTIVE: To compare repeated freeze-thaw and ultrasonication for the collection of lysate of MSCs. METHODS: Adipose-derived mesenchymal stem cells (ADMSCs) were isolated from the abdominal subcutaneous fat of healthy individuals, and purified with adherence screening method, followed by in vitro amplification using fetal bovine serum medium. The common surface makers of these cells were tested by flow cytometry (1×109, 2×109, 4×109/L). Repeated freeze-thaw and ultrasonication were employed for cell cytoclasis at three different densities respectively in saline and double distilled water, and a comprehensive comparison was performed on cytoclasis rate and the content of protein in cell lysate between the two methods. RESULTS AND CONCLUSION: (1) ADMSCs obtained from in vitro isolated human adipose tissue grew in a swirl or radial pattern with a homogenous size and neat arrangement. CD44, CD90, CD105 and other commonly used surface markers were highly expressed. (2) The study for optimization of lysate collection revealed that the higher cell density implicated a longer time for cell wall disruption and cytoclasis, as well as significantly increased cytoclasis rate. (3) BCA protein assay showed that the highest content of protein was obtained in saline solvent using ultrasonication method. Comprehensive analysis on the results leads to a conclusion that ultrasonication method with saline as the solvent is the optimized method for extraction of ADMSCs lysate, and the cell concentration of less than 4×109/L is recommended.

Objective To investigate some improvements in the surgical techniques of adult-to-adult living donor liver transplantation( A-A LDLT) without the middle hepatic vein(MHV) for hepat-ic vein reconstruction. Methods The retrospective analysis was made on the clinical data of 11 recipi-ents who underwent the operation in A-A LDLT including the hepatic vein reconstructed in right liver lobe without MHV from June 2007 to January 2008. The key techniques included reconstructing out-flow of graft on shaping the tips of vena cava and right hepatic veins, cadaveric vein allografts stored in 4℃ UW solution within 7d being used for significant-sized hepatic vein reconstruction such as tributa-ries of the middle hepatic vein from V5, V8 and right inferior hepatic vein. Results 10 cases success-fully underwent reconstruction of outflow of graft on shaping the tips of vena cava and right hepatic veins and the outflow reconstruction ratio of V5, V8 and right inferior hepatic vein was 81. 8% (9/11), 7 one-vein reconstruction, 1 two-vein reconstruction and 1 three-vein reconstruction. 1 recipient died of renal failure and pulmonary infection 14 days after operation without venous outflow obstruc-tion. Doppler ultrasonography showed no thrombosis and the blood flowed smoothly in the right he-patic vein of other 8 recipients during the 9th to 15th mouth of follow-up. The cumulative patency rates of these 8 survivals for interposition vein grafts were 100% (11/11), 72. 7 %(8/11), 54. 5%(6/11) and 36. 5%(4/11) in 1, 3, 6 and 9 mouths, respectively. The regeneration of paramedian sectors was equivalent. Conclusion Shaping the tips of vena cava and right hepatic veins and using cadaveric vein allografts in adult-to-adult right lobe living donor liver transplantation for hepatic vein reconstruc-tion are both safe,simple and effective methods.This approach can be recommended.

Objective To study the directional migration of bone rnarrow-derived stenl cells (BMSCs)towards the glioma and the distribution of tbe migrated BMSCs in the brain. Methods In vitro cultured BMSCs isolated from the bone marrow of male Wistar rats were identified using immunofluorescence technique. and their stem cell properties wcrc assessed by means of induced differentiation in vitro into adipocytes and osteoblasts.Wistar rat models bearing glioma were established by stereotactic injection of C6 glioma cells into the basal ganglia,and 7 days later.Brdu-labeled BMSCs werc injected stereotaetically into the contralateral hemisphere or the contralateral ventricle, or intravenously via the tail vein.The rats were sacrificed 14 days afterthe BMSC injection,and coronal paraffin sections(5 μm thick)of the brain tissue were prepared.HE staining was used to observe the extent of intracranial glioma growth,and the distribution of the BMSCs in the glioma tissue as well as in the brain tissue was identified With immunofluorescent staining. ResuIts The BMSCs implanted into either the contrahteral hemisphere or the contralateral ventricle were found to migrate towards the glioma mostly along the needle tract,and distinct green fluorescence emitted by the labeled BMSCs Was detected on the boundary between the glioma and the normal brain tissue.A few fluorescent BMSCs were also seen around the glioma tissue after intravenous injection of the cells. Conclusion BMSCs injected into the cerebral parenchyma,contralateral ventricle,or the tail vein are capable of directional migration into the glioma tissue,suggesting their potential as a new vehicle for delivering therapeutic genes into the glioma tissue.

To study the proliferation characteristics of PPV in differently infected way and the variance of concentrations in different cells. A strain of porcine parvovirus(PPV) was adapted to PK-15 cells, and a Real-time fluorescent quantitative PCR (FQ-PCR) assay was developed based on the specific region of the NS1 gene of PPV to quantify the PPV. The FQ-PCR was used to measure the viral concentration of virus-infected cells by simultaneous or step by step inoculation and plot one-step growth curves. The proliferation characteristics of PPV strain in different cells lines (HeLa, MDBK, PK-15 ,ST, F81, BHK-21 and Marc-145) was also compared. The results showed the PK-15 cell -adapted strain of PPV produced CPE after 12 passages, and maintained stable CPE at the following 10 messages. The one-step growth curve showed that the virus concentration of simultaneous inoculation was higher than that of the step-by-step inoculation, and the proliferation cycle of step-by-step inoculation was shorter. The proliferation ability of PPV strain in different cells showed that CPE appeared first inPK-15, followed by ST, HeLa and MDBK, and the virus concentration was highest in ST, followed byPK-15, MDBK and HeLa. NO proliferation was observed in F81, BHK-21 and Marc-145 cells. These findings lay a material foundation for the basic researches on PPV and the development of vaccine.
Animals ; Cell Line ; Cricetinae ; Cytopathogenic Effect, Viral ; DNA, Viral ; genetics ; Female ; Haplorhini ; Humans ; Male ; Parvoviridae Infections ; virology ; Parvovirus, Porcine ; genetics ; physiology ; Real-Time Polymerase Chain Reaction ; Reproducibility of Results ; Sensitivity and Specificity ; Swine ; Viral Proteins ; genetics ; Virus Replication