To study the effect of HBx gene on the apoptosis of the cell lines (L02, HepG2) and the interaction between HBx and X-linked inhibitor of apoptosis protein (XIAP), the apoptosis of pcDNA3.1-HBx transiently transfected cell lines (L02, HepG2) was detected by flow cytometry and the mRNA expression of XIAP was assayed by real-time RT-PCR. Our study showed (1) the morphology of L02/pcDNA3.1-HBx was changed and the appearance of the cells mimicked that of HepG2 cells; (2) HBx gene could be detected in L02/pcDNA3.1-HBx and HepG2/ pcDNA3.1-HBx; (3) the apoptosis rate of L02/pcDNA 3.1-HBx was higher than that of L02 cells (P<0.01) and the apoptosis rate of HepG2/pcDNA3.1-HBx was lower than that of HepG2 cells (P<0.05); (4) the XIAP expression in L02 was about 3 times that in L02/pcDNA3.1-HBx cells (P<0.01), and the expression of XIAP in HepG2/pcDNA3.1-HBx was about 4 times that in HepG2 (P<0.01). It is concluded that HBx gene may promote the apoptosis of normal hepatocytes and inhibit the apoptosis of cells of hepatic carcinoma by regulating the expression of XIAP.

This study is to investigate the effect of Vam3, a dimeric derivative of resveratrol, on SNP-induced apoptosis and its potential mechanism in rat articular chondrocytes. Isolated rat articular chondrocytes were treated with sodium nitroprusside (SNP), a NO donor, to induce apoptosis. Apoptosis percentage was evaluated by Annexin V-PI and nucleus fracture was examined by DAPI staining. Level of intracellular reactive oxygen species (ROS) was detected using 2, 7'-dichlorofluorescin diacetate (DCFH-DA) as a fluorescence probe by fluorescence microplate reader. The change in mitochondrial membrane potential was detected by TMRE staining. Expressions of SIRT1, acetylated p53 (ac-p53), cleaved caspase 9 and cleaved caspase 3 were determined by Western blotting. It showed that Vam3 up to 10 micromol x L(-1) could significantly reduce SNP-induced rat articular chondrocytes apoptosis (P < 0.01) and nucleus fracture, inhibit the increase of intracellular ROS level (P < 0.01) and reverse the decrease in mitochondrial membrane potential (P < 0.01). Simultaneously, Vam3 could upregulate the expression of SIRT1, deacetylate p53, and inhibit the cleavage of caspase 9 and caspase 3 (P < 0.01) of rat articular chondrocytes exposed to SNP. This study indicates Vam3 could protect rat articular chondrocytes against SNP-induced apoptosis, perhaps through the upregulation of SIRT1 and deacetylation of p53.
Animals ; Apoptosis ; drug effects ; Arabidopsis Proteins ; pharmacology ; Cartilage, Articular ; cytology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cells, Cultured ; Chondrocytes ; cytology ; metabolism ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Nitric Oxide Donors ; antagonists & inhibitors ; pharmacology ; Nitroprusside ; pharmacology ; Qa-SNARE Proteins ; pharmacology ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Sirtuin 1 ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective:To observe the efficacy and safety of intrapleural injection with urokinase ( UK) in the treatment of tubercu-lous pyothorax. Methods:Totally 120 patients with tuberculous pyothorax were randomly divided into two groups. The vomicae in the control group were rinsed with 5% sodium bicarbonate, and those in the observation group were treated with UK. The total effective rate, symptom relief time, closure time of vomicae, pleural thickness changes and adverse reactions were observed in the two groups. Results:The total effective rate of the control group and the observation group respectively was 81. 7% and 98. 3%, which showed sta-tistically significant difference (P<0. 01). The symptom remission time and the pus cavity closure time of the observation group were shorter than those of the control group (P<0. 05). The pleural thickness of the observation group was thinner than that of the control group at the end of the treatment (P<0. 05). The incidence of adverse drug reactions in the observation group was significantly higher than that in the control group (P<0. 01). Conclusion:The curative effect of UK in the treatment of tuberculous pyothorax is better than that of sodium bicarbonate, however, it is necessary to pay attention to monitoring such adverse reactions as bleeding, low heat and the others.

Objective To analyze the CT findings of melamine induced urinary calculi.Methods Nineteen children with a history of ingestion of melamine contaminated infant formula milk were studied, including 12 males and 7 females, age ranged from 50 days to 5 years.Results CT demonstrated renal pelvic and ureteral stones in 13 cases, with urinary obstruction in 9 of them.The size of the stones ranged from 0.3 cm × 0.3 cm to stag-horn calculus.The density of the stones measured from a low of 40-70 HU up to a high of 410 HU with an average density of 160 HU.Conclusion CT scan is an excellent modality in demonstrating urinary tract calculi caused by melamine. It is the method of choice when ultrasound examinations are equivocal.

Objective To investigate the cellular immune response to the recombinant DNA vaccine CRT120/HPV6bE7 in mice. Methods The recombinant encoding HPV6bE7, linked with CRT120, was constructed in pcDNA3.1 eukaryotic vector with the report gene GFP. The pcDNA3.1-GFP -HPV6bE7 was transfected into B16 cells by a lipofectamine kit. The C57BL/6 mice were vaccinated with recombinant DNA plamids. The T-cell phenotype in the peripheral blood lymphocytes of the immunized mice was measured by flow cytometry. The CTL activity of the lymphocytes from the spleens and lymph nodes, and the levels of IL-2 and IFN-? were analyzed. Results The constructed recombinant plasmid CRT120/HPV6bE7 was analyzed. Positive transfected cell clones were established and could stably express the target gene HPV6bE7. Compared with HPV6bE7, CRT120/HPV6bE7 plasmid enhanced to a greater extent CD8+ T-lymphocyte differentiation, the number of TCR?? T-lymphocytes and the levels of IL-2 and IFN-? (all P

BACKGROUND:Titanium and titanium aloy are used mostly in artificial joints, fracture fixation, and oral transplantation, while there are complex cases of insufficient bone mass in these areas. The deepened research of stem cels offers a solution for bone injury to promote new bone formation. The biocompatibility of titanium and stem cels and optimization of titanium surface modification have aroused people's attention. OBJECTIVE:To investigate whether the biocompatibility of titanium and human adipose-derived mesenchymal stem cels can be improved by type I colagen modification of titanium sheets. METHODS:The experiment was divided into two groups. Modification group: titanium sheet was modified with type I colagen; control group: titanium sheet was not modified with type I colagen. Human adipose-derived mesenchymal stem cels at passage 6 were implanted into titanium sheet in two groups. Then we calculated the number of adherent cels in two groups at 1, 2 and 4 hours after implantation, and compared the celladhesion rate. MTT assay was used to observe the proliferation of cels on titanium sheet at 2, 4, 6 and 8 days after implantation. DNA and protein content of cels were detected at 3, 6, 9 days after implantation. The growth of human adipose-derived mesenchymal stem cels seeded upon the titanium sheets was observed under scanning electron microscope at 6 days. RESULTS AND CONCLUSION:When the cels were cultured for 1 hour and 2 hours, the number of adherent cels in the modification group was higher than in the control group (P < 0.05). The absorbance of cels in two groups was increased as the culture time, as detected by MTT assay. The modification group had a significantly higher absorbance value than the control group at 4, 6, 8 days (P < 0.05). DNA and protein contents of the cels in the modification group were higher than that in control group at 6 and 9 days (P < 0.05). At 6 days, the number of adherent cels and secretion of adherent stromal cellmatrix in the modification group were significantly better than that in control group, observed by scanning electron microscopy. Type I colagen modified titanium sheets have good surface activity and biocompatibility, and can promote the proliferation of human adipose-derived mesenchymal stem cels.

BACKGROUND:The safety and efficacy of simultaneous bilateral total knee replacement or selective unilateral total knee arthroplasty in patients with severe osteoarthritis of the knees are stil controversial. OBJECTIVE:To compare safety and clinical efficacy of patients with osteoarthritis knees after simultaneous bilateral total knee replacement or selective unilateral total knee replacement. METHODS:Total y 60 cases with severe osteoarthritis of the knees (90 knees) undergoing total knee replacement were divided into unilateral total knee replacement group (n=30, 30 knees), and the simultaneous bilateral total knee replacement group (n=30, 60 knees). RESULTS AND CONCLUSION:There was no significant difference in the incidence of other complications such as infection, mortality, pulmonary embolism in patients of both groups (P>0.05). The incidence of cardiovascular complications, postoperative blood loss and blood transfusion were higher in the bilateral knee group than in the unilateral knee group (P<0.05). During fol ow-up at 1 year after replacement, no significant differences in range of motion, muscle strength of quadriceps and hospital for special surgery knee score were detected in patients of both groups (P>0.05). However, Visual Analogue Scale scores were significantly lower in the bilateral knee group than in the unilateral group (P<0.05). These data indicated that the risk of cardiovascular complications was high in patients receiving bilateral total knee replacement. Patients with severe cardiovascular disease should avoid simultaneous bilateral total knee arthroplasty.

Objective: To study the nutritional status and energy metabolic characteristics of patients with nonalcoholic fatty liver disease (NAFLD), and to provide evidence for clinical evaluation and intervention. Methods: A total of 359 NAFLD patients diagnosed on ultrasound from June 2015 to March 2017 were selected as study subjects and divided into mild, moderate to severe fatty liver disease group and 50 healthy subjects as control group. The changes of ICW, ECW, body fat, skeletal muscle, protein and visceral fat area (VFA) of patients and controls were analyzed by using body composition analyzer. The energy metabolism index was measured by the oxidation rate of resting energy expenditure(REE), respiratory quotient (RQ), and the oxidation rates of the three nutrients (CHO %, FAT %, and PRO %). According to different types of data, non-parametric tests like Kruskal-Wallis or χ² were used for this analysis. Results: Compared with the mild fatty liver group and the control group, the moderate and severe fatty liver group the BMI, waist circumference, waist-hip ratio were significantly elevated (P-value < 0.001), and their serum alanine aminotransferase, triglyceride, total cholesterol, high-density lipoprotein, low-density lipoprotein, FBS levels were significantly increased (P value < 0.05). The Body composition analysis showed that there was no significant difference in skeletal muscle content between the three groups (P = 0.067). The ICW, ECW, protein, body fat content of moderate and severe fatty liver group were significantly higher than those of mild fatty liver group and control group (P < 0.01), but there was no significant difference between the mild fatty liver group and the control group. There was significant difference in the VFA between the three groups, while VFA in the moderate and severe fatty liver group was significantly increased. Metabolic results showed that the RQ of patients with moderate-severe fatty liver and mild fatty liver were 0.72 ± 0.08 and 0.78 ± 0.06, respectively, which were lower than those of the control group (0.80 ± 0.02), P = 0.004. Resting energy expenditure (REE) was not significantly different between moderate and severe fatty liver group and mild fatty liver group (P = 0.207), but both were significantly higher than those of the control group (P < 0.001). The percentages of CHO, FAT and PRO in moderate and severe fatty liver group were 19.49% ± 9.71%, 66.23% ± 12.54% and 14.22% ± 6.11% respectively. Compared with the control group, CHO % decreased, and FAT % increased. Conclusion NAFLD patients have different extent of nutritional imbalance and energy metabolism disorders, the use of Body Composition analyzer and metabolic cart can comprehensively assess and monitor NAFLD patient’s nutrition and energy metabolism status, to provide a basis for clinical intervention.

OBJECTIVETo study the chemical constituents from rhizome of Pulsatilla dahurica.

METHODThe constituents were isolated and purified by various chromatographic methods. AR compounds were identified on the basis of spectral analysis and physico-chemical characters.

RESULTSix compounds were isolated from the 70% alcohol extract of the rhizome identified as hederagenin ( I ), hederagenin 3-O-alpha-L-arabinopyranoside (II), hederagenin 3-O-beta-D-glucopyranosyl(1-->2)-alpha-L-arabinopyranoside (III), hederagenin 3-O-beta-D-glucopyranosyl(1 -->2) [beta-D-glucopyranosyl(1-->4)]-alpha-L-arabinopyranoside (IV), beta-sitosterol (V) and daucosterol (VI), respectively.

CONCLUSIONCompounds I approximately VI were isolated from this plant for the first time.

Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Pulsatilla ; chemistry ; Rhizome ; chemistry ; Saponins ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification

The aim of this study is to investigate the anti-inflammatory effect of the adenosine derivative N6-(3-hydroxylaniline) adenosine (WS070117M1) on cigarette smoke plus LPS (lipopolysaccharide)-induced chronic obstructive pulmonary disease (COPD) in mice and its mechanism. COPD model was established by exposing male BALB/c mice to cigarette smoke and challenged with LPS inhalation. Supernatants of bronchoalveolar lavage fluid (BALF) were harvested and IL-1β, IL-6, IL-8 and TGF-β1 levels were measured by ELISA (enzyme-linked immunesorbent assay). The number of total white blood cells and neutrophils in bronchoalveolar lavage fluid was counted separately. Lung tissue was stained with Mayer 's hematoxylin and eosin for histopathologic examination. pAMPKa protein expression and distribution of lung tissue were analyzed by immunohistochemistry method. In vitro, levels of AMPKα phosphorylation in phorbol-12- myristate-13-acetate (PMA) differentiated THP-1 cells was detected by immunohistochemistry, IL-8 level in supernatants of cigarette smoke condensate stimulating PMA differentiated THP-1 cells was measured by ELISA. The results showed that WS070117M1 treatment significantly activated AMPKa in the lung tissue. It also resulted in down regulation of IL-1β, IL-6, IL-8 and TGF-β1 levels in bronchoalveolar lavage fluid and IL-8 level in cigarette smoke condensate stimulating PMA differentiated THP-1 cells. In addition, WS070117M1 could inhibit the recruitment of total white blood cells and neutrophils. These results suggest that WS070117M1 may alleviate the airway inflammation by activating AMPK in the lung tissue.
AMP-Activated Protein Kinases ; metabolism ; Adenosine ; analogs & derivatives ; Animals ; Bronchoalveolar Lavage Fluid ; Cell Line, Tumor ; Disease Models, Animal ; Humans ; Inflammation ; drug therapy ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Leukocyte Count ; Lipopolysaccharides ; Male ; Mice ; Mice, Inbred BALB C ; Neutrophils ; cytology ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Smoke ; adverse effects ; Tobacco ; Transforming Growth Factor beta1 ; metabolism