Dual-energy X-ray absorptiometry(DEXA),scanning electron microscopy,and bone biomechanical test were used to assess comprehensively bone quantity and quality of ovariectomized rats. In OVX rats,not only bone mineral density (BMD) of lumbar vertebrae in vivo and vitro,but also BMD of femora (except for R3 region) and proximal metaphysis(R1 region) in vitro decreased obviously (P<0.01)，whose bone loss rates of L5 and L6 were the highest and achieved 13%; the trabeculae of OVX rats were few，fine, and discontinued and there were lacunae on the surface;in OVX rats,both compressive strength of vertebral bodies and the mechanical properties of femora decreased;the falling degree of the former was greater;the maximal compressive power of lumbar vertebrae decreased with 33.32%. We conclude that after 4-month post-ovariectomy, the bone quantity and quality of six-month-old rats decreased, especially in the area of abundant cancellous bone such as vertebral bodies,distal femora; the assessment of the efficiency of new drugs to prevent and treat postmenopausal osteoporosis using rat models should include these drugs' effects to the bone mass, the bone structure, and the bone strength.

Serum osteoprotegerin (OPG) concentration was measured in 647 healthy female adults (aged 20-81 years), and was compared with that of other races. The serum OPG was positively correlated with age (r = 0.276, P <0.01). The geometric mean value of serum OPG in premenopausal women was significantly lower than those in perimenopansal and postmenopausal women. The serum OPG in middle-aged Chinese women was signifieandy higher than those in middle-aged Austrian and Icelandic, but this was quite contrary to the results obtained in old-aged women.

Objective To explore the molecular mechanism of glucocorticoid (GC)-induced osteoporosis (GIOP). Methods Thirty-two female SD rats after matching body weight were divided randomly into three groups: baseline group (n = 10), control group (n = 11) and GC-treated group (n = 11). The administration time was 9 weeks. Bone mineral density (BMD) was measured with dual energy X-ray absorptiometry. A high resolution micro-CT was used to quantify the densitometric and microarchitectural properties of trabeculae in the proximal metaphysis of right tibia. In situ hybridization histochemistry and immunohistochemistry were used to detect the expression of cannabinoid type 1 receptor (CBI R) in the proximal metaphysis of left tibia. Results At the end of the experiment, whole-body BMD in vivo in the control group [(0. 156±0. 008) g/cm2]was higher than that in the baseline group [(0.147±0.006)g/cm2], while the whole-body BMD in vivo [(0.147±0.006) g/cm2]and total BMD in vitro at femurs in the GC-treated group [(0.220±0.011) g/cm2]was lower than those in the control group [(0. 240±0. 024)g/cm2]. Compared with the baseline group and control group, there was a remarkable decrease in the volumetric BMD, tissue BMD, trabecular number and trabecular connectivity (P<0.05) in the GC-treated group, while there was a significant increase in trabecular separation (P < 0. 05) and trabecular thickness also increased in the proximal metaphysis of tibiae in the GC-treated group. The expression level of CB1R mRNA and protein in osteoclasts in the GC-treated group was markedly higher than that in the baseline group and control group (P < 0. 05). There was a close correlation between the expression level of CB1R mRNA, protein in osteoclasts and some microarchitectural parameters in the proximal metaphysis in the GC-treated group (P<0.05). Conclusions The administration of GC is associated with a decrease in BMD and deterioration in microarchitecture of trabecular bone in rats tibiae. Glucocorticoid may up-regulate the CB1R expression level in osteoclasts and this may be a kind of molecular mechanism of GIOP.

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.

Atropa belladonna L. is the officially medicinal plant species and the main commercial source of scopolamine and hyoscyamine in China. In this study, we reported the simultaneous overexpression of two functional genes involved in biosynthesis of scopolamine, which respectively encoded the upstream key enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53) and the downstream key enzyme hyoscyamine 6beta-hydroxylase (H6H; EC 1.14.11.11) in transgenic hair root cultures of Atropa belladonna L. HPLC results suggested that four transgenic hair root lines produced higher content of scopolamine at different levels compared with nontransgenic hair root cultures. And scopolamine content increased to 8.2 fold in transgenic line PH2 compared with that of control line; and the other four transgenic lines showed an increase of scopolamine compared with the control. Two of the transgenic hair root lines produced higher levels of tropane alkaloids, and the content increased to 2.7 fold in transgenic line PH2 compared with the control. The gene expression profile indicated that both PMT and H6H expressed at a different levels in different transgenic hair root lines, which would be helpful for biosynthesis of scopolamine. Our studies suggested that overexpression of A. belladonna endogenous genes PMT and H6H could enhance tropane alkaloid biosynthesis.

OBJECTIVETo study the effects of Chaiqiyigan Granula (CG) and Taxol on the growth of hepatocellular carcinoma (HCC) xenografts and expression of Bax, p53 and VEGF in nude mice.

METHODSWhole-body fluorescence imaging was used to visualize the growth of HCC in nude mice bearing hepG2/EGFP cell xenograft. Immunohistochemistry was used to determine the content of Bax, p53 and vascular endothelial growth factor (VEGF) in the tumor tissues.

RESULTSCompared with normal saline, Taxol alone and in combination with CG significantly inhibited the growth of HCC xenografts in the nude mice. The combined treatment with CG and Taxol produced a stronger inhibitory effect on the tumor growth than Taxol alone in the third and fourth weeks. The volume and weight of the xenografts were decreased in the combined treatment group compared with those in saline treatment group. CG combined with Taxol increased the expression of Bax and reduced the expression of p53 and VEGF in the tumor xenografts.

CONCLUSIONCG can enhance the inhibitory effects of Taxol on the growth of HCC xenografts, and this effect is related to the up-regulation of Bax and down-regulation of p53 and VEGF expression in the tumor.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Mice ; Mice, Nude ; Optical Imaging ; Paclitaxel ; pharmacology ; Tumor Suppressor Protein p53 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism

Objective To explore the effect of Ginkgo biloba extract (EGb761) on apoptosis of K-ras mutational human colon cancer cells DLD1(DLD1/G13D)and its mechanism. Methods Human colon cancer cell lines DLD1/G13D and DLD1 with K-ras wild type(DLD1/WT)were cultured in vitro,the cell proliferation and apoptosis after 24 h of EGb761 were measured. Proteins involved in related signal pathway were detected by Western blot or ELISA. Results EGb761 reduced cell proliferation and induced cell apoptosis in a concentration-dependent manner in DLD1/WT and DLD1/G13D cells. EGb761 downregulated the expression of RIP1, impaired the phosphorylation of IκB and decreased the level of NF-κB in DLD1/WT and DLD1/G13D cells[DLD1/G13D: (24±4)%, DLD1/WT: (29±9)%(P<0.05). Conclusion EGb761 restrains the proliferation and induces the apoptosis of DLD1/WT and DLD1/G13D cells. The mechanism may be related to the degradation of RIP-1 and inhibition of activation of NF-κB signaling pathway.

Objective To analyze the clinical characteristics and explore the risk predictors on mortality in elderly patients with acute cholecystitis and cholangitis.Methods We conducted a retrospective analysis of elderly patients hospitalized in the Second Medical Center of General Liberation Army Hospital for acute cholecystitis and cholangitis during 2000 to 2018.Clinical data and risk predictors on mortality were assessed.The patients were stratified into three groups based on age:Ⅰ (65-74 years old),Ⅱ (75-84 years old),and Ⅲ (≥85 years old).Logistic regression analysis was used to identify the predictors of mortality.Results A total of 574 patients were finally enrolled with the mean age 87.6 years including 191 in group Ⅰ,167 in group Ⅱ,and 216 in group Ⅲ.The main cause of acute cholecystitis and cholangitis was gallstone (76.3％),and the main symptom was abdominal pain (62.9％),followed by chills(62.5％),fever(59.8％),jaundice (47.2％) and septic shock(26.3％).Cholecystitis was the most common diagnosis in groups Ⅰ and Ⅱ,whereas it was cholangitis in group Ⅲ.Percutaneous transhepatic biliary/gallbladder drainage (PTBD/PTGD) and endoscopic retrograde cholangiopancreatography (ERCP) were administrated more frequently in groups Ⅲ.A total of 35 patients (6.1％) died during follow-up.Senior in age (OR=11.1),the Charlson comorbidity index (OR=19.5),cancers (OR=9.6),blood stream infections (OR=7.4),severity of cholecystitis and cholangitis (OR=4.2)were risk factors associated with mortality.Conclusions Even in the elderly patients with acute cholecystitis and cholangitis,comorbidity is one of the main factors affecting clinical outcomes.Due to the poor performance,this group of population presents more severe disease and undergoes conservative treatment strategies.

It was reported that pancreatic arteries constricted during the early phase of bile salt-induced acute pancreatitis (AP), leading to pancreatic microcirculatory disturbance. We conducted this experiment to verify whether the above-mentioned finding was true. AP was induced with intraductal injection of taurodeoxyholate. Small pancreatic artery pressure in dogs was recorded. Functional capillaries were counted and calibrated by multiplying wet weight of pancreas. Pancreatic perfusion was measured with Laser Doppler flowmeter. Pancreatic arterioles of rats dilated during the initial 20 min of AP, and pancreatic arterial pressure declined during the early phase of AP in dogs (from 104.5 +/- 4.8 mmHg to 54.6 +/- 5.6 mmHg). The hematocrit of blood from inferior vena cava was significantly lower than that of portal vein at 5 min after pancreatitis induction. The "true" pancreatic functional capillary density increased. The early pancreatic microcirculatory disturbance coincided with a marked increase of portal vein pressure (PVP) as high as 9.18 +/- 0.78 mmHg. Reduction of PVP to baseline level was followed by a marked increase of pancreatic perfusion (by 1.4-fold). Arterial dilatation, but not constriction, occurred during the early phase of bile salt-induced AP. The pancreatic microcirculatory disturbance was due to a marked rise in PVP that greatly reduced the pressure difference in the pancreatic blood vessels and increased plasma extravasation which led. to local hemoconcentration.
Animals ; Bile Acids and Salts ; adverse effects ; Hypertension, Portal ; complications ; Male ; Microcirculation ; drug effects ; physiology ; Pancreas ; blood supply ; Pancreatitis ; etiology ; physiopathology ; Portal Pressure ; Portal Vein ; physiopathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the mechanism of lumen loss of the left circumflex ostium after main vessel stent implantation.

METHODSTwenty-eight patients undergoing provisional T technique were enrolled in this study. Intravascular ultrasound (IVUS) examination was performed before and after main vessel stenting and kissing balloon post-dilatation to evaluate the geometrical changes of the vessels.

RESULTSThe CSA of LCX ostium lumen decreased significantly from 5.9∓2 mm(2) to 4.9∓1.9 mm(2) (P<0.01) after the procedure, and the CSA of LCX ostium P and M increased from 5.4∓2.9 mmmm(2) to 5.7∓2.9 mm(2) (P=0.21) after the main vessel stenting. The changes in LCX ostium lumen CSA was correlated with the changes of LCX ostium EEM CSA but not the LCX ostium P and M CSA. After kissing balloon post-dilatation, the CSA of LCX ostium lumen increased from 4.9∓1.9 mm(2) to 5.5∓1.9 mm(2) (P<0.01) , and the CSA of LCX ostium P and M showed no obvious changes (5.7∓2.9 mmmm(2) vs 5.7∓2.6 mmmm(2), P=0.89). The changes of LCX ostium lumen CSA were correlated with the those of the LCX ostium EEM CSA (R=0.432, P=0.02).

CONCLUSIONAfter stent implantation from the LMCA to the LAD, most of lumen losses of the LCX are due to carina shift, and in occasional cases, plaque shift occurs from the distal LMCA to the ostium of the LCX. Kissing balloon technique can adjust carina shift but can not improve plaque shift.

Aged ; Angioplasty, Balloon, Coronary ; Coronary Artery Disease ; diagnostic imaging ; therapy ; Coronary Stenosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Stents ; Treatment Outcome ; Ultrasonography, Interventional