Plasma levels of artial natriuretic polypeptide (ANP), cyclic GMP (cGMP), renin activity (PRA), angiotensin II (AT II) and arginine vasopressin (AVP) were measured by radioimmunoassay in 30 patients with acute heart failure (AHF), 30 chronic heart failure (CHF), 30 chronic atrial fibrillation (CAF) and 27 paraxysmal atrial fibrillation and supraventricular tachycardia (SVT). The results showed that plasma ANP and cGMP levels in all these four groups were significantly higher than those in the normal group (P

OBJECTIVETo investigate the influence of Rehmannia glutinosa oligosaccharides (ROS) on the proliferation of 3T3-L1 adipocytes and insulin resistance.

METHOD3T3-L1 preadipocytes were cultured, the proliferation of 3T3-L1 preadipocytes was detected by MTT method. Insulin resistant 3T3-L1 adipocytes cell model was induced by dexamethasone and the change of glucose concentration in cell culture was determined after ROS treatment.

RESULTIn the high glucose DMEM culture media, MTT method showed that the absorbance at 570nm of 3T3-L1 preadipocytes was increased and that of 3T3-L1 adipocytes was decreased. ROS significantly increased glucose consumption in 3T3-L1 preadipocytes and adipocytes culture in a concentration-dependent manner. ROS improved the sensitivity of 3T3-L1 adipocytes to insulin.

CONCLUSIONROS can promote the proliferation of 3T3-L1 preadipocytes, inhibite the proliferation of 3T3-L1 adipocytes, and also, significantly improve insulin resistance induced by dexamethasone.

3T3-L1 Cells ; Adipocytes ; cytology ; Animals ; Cell Proliferation ; drug effects ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Insulin Resistance ; Mice ; Oligosaccharides ; administration & dosage ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rehmannia ; chemistry

AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-SMA in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated siRNA targeting CTGF.METHODS: HLECs were transfected with small interfering RNA (siRNA) targeting CTGF,labeled with 5`-fluorescein isothiocyanate (5`-FITC) and coated with lipofectamine.The transfection ratio was evaluated via fluorescence intensity.Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs.Quantitative RT-PCR,cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine.The transfection ratio reached 95% at 24h.The proliferation of HLECs was inhibited by siRNA after 72h transfection.The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h.This effect was not found in negative control siRNA.CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs,which is a potential therapeutic strategy for posterior capsular opacification.

Objective Pemetrexed and β-elemene can inhibit the growth of tumor cells .This study was to investigate the effect of pemetrexed combined with β-elemene on the proliferation and apoptosis of cervical cancer HeLa cells. Methods Cervical cancer HeLa cells were treated with pemetrexed at the concentrations of 38, 76, 152, 228, and 304μg/mL, and at 24 and 48 hours of treatment subjected to MTT for detection of their proliferation .The experiment included four groups , with the cells treated with β-elemene ( 125μg/mL) , pemetrexed ( 76 μg/mL ) , β-elemene ( 125 μg/mL ) +pemetrexed (76μg/mL), and nothing (blank control) for 24 hours, followed by determination of their proliferation and apoptosis by MTT and flow cytometry, respectively. Results Pemetrexed at 38, 76, 152 and 228μg/mL inhibited the proliferation of the HeLa cells in a concentration-dependent manner, with the inhibition rates of (7.24 ±3.78), (7.94 ±4.37), (11.10 ±2.86) and (15.88 ± 3.38)%at 24 hours, and (16.69 ±0.95), (22.54 ±1.53), (24.48 ±0.92) and (25.54 ±3.61)%at 48 hours, both with statis-tically significant differences between any two groups (P<0.05).Significant differences were also found in the proliferation rate of the same concentration of pemetrexed at the two time points (P<0.05).The combination of pemetrexed and β-elemene showed an inhibi-tion rate of (49.95 ±5.76)%at 24 hours, remarkably higher than (24.36 ±5.59)%in theβ-elemene group and (10.69 ±1.37)%in the pemetrexed group (P<0.01). Conclusion Pemetrexed combined with β-elemene can significantly inhibit the proliferation and synergistically accelerate the apoptosis of HeLa cells .

Objective To investigate the initiation pathway of corneal endothelial cell apoptosis induced by high-pression.Methods Primary rabbit corneal endothelial cells were identified by immunohistochemistry and cultured under high pressure 50 mmHg (1 kPa =7.5 mmHg) for 1 h,2 h,24 h,respectively,while cells cultured under the normal pressure 15 mmHg served as the normal pression group.In addition,the first generation of rabbits corneal endothelial ceils with 70％ to 80％ fusion were pretreated with 10-6 mol · L-1 anti-Caspase 8 and anti-Caspase 9 for lh,followed by 50 mmHg pression for the treatment of the cells;while cells cultured with no inhibitor in the same pression served as the control group.Then the expression of P53 and Bcl-2 protein was detected by Western blot,and cytochrome C in rabbit corneal endothelial cells was determined by immunofluorescence staining in all groups.Results The expression levels of P53 in the 50 mmHg group were 0.651 +0.007,0.805 ±0.006 and 0.839 ±0.011 after 1 h,2 h,24 h high-pression respectively,which were significantly higher than those in the normal pressure group (0.033 ± 0.004),and the difference approached statistical significance (all P ＜ 0.01).The expression of P53 protein in corneal endothelial cells gradually increased as time went on,and the difference was statistically significant between each two time-points (all P ＜ 0.01).Moreover,the expression of Bcl-2 in the 50 mmHg pressure group was 0.590 ± 0.009,0.724 ± 0.005 and 0.34 ± 0.016,respectively,which was higher than that in the normal pressure group (0.081 ±0.013),with signifi cant difference (all P ＜ 0.01),and the difference approached statistical significance between each two time points in this group (all P ＜ 0.01).The expression level of P53 in anti-Caspase 9 and anti-Caspase 8 group was 0.535 ± 0.007 and 0.703 ± 0.010,respectively,which was significantly lower than that in the control group (0.727 ± 0.021),and the difference was statistically significant (all P ＜ 0.01).The expression of Bcl2 was 0.312 ± 0.003 and 0.442 ± 0.011,respectively,which were significantly lower than that in the control group (0.501 ± 0.011),with statistical difference (P ＜ 0.01).Finally,the expression of P53 and Bcl-2 in anti-Caspase 9 group was lower than that of anti-Caspase 8 group (P ＜ 0.01),indicating that anti-Caspase 9 had more enhanced inhibitory effect on the apoptosis of corneal endothelial cells than anti-Caspase 8.Conclusion AntiCaspase 9 inhibitor could effectively block the corneal endothelial cell apoptosis induced by high pressure.And the damage from high pressure on corneal endothelial cells mainly triggers the release of cytochrome C from chondriosome to activate the endogenous enzyme linked apoptotic pathway in which Caspase 9 involves.

OBJECTIVETo investigate the effects of Rehmannia glutinosa oligosaccharides (ROS) on the proliferation of HepG2 and insulin resistance.

METHODThe HepG2 cells were divided into control group, rosiglitazone (3.4 mg x L(-1)) treated group and ROS (0.1-30 mg x L(-1)) treated group. The proliferation of HepG2 was detected by MTT method. Insulin resistant HepG2 cells model was induced by high concentration of insulin, then the effects of ROS on glucose consumption in insulin resistant HepG2 cells were investigated.

RESULTIn the middle glucose culture medium, the absorbance at 570 nm of HepG2 was increased by high concentration of ROS, and decreased by low concentration of ROS by using MTT method, a concentration-dependent manner. ROS increased glucose consumption in HepG2 cells, and showed a better effect at the dose of 10 mg x L(-1). ROS promoted the glucose consumption in insulin resistance of HepG2 cells, improved the sensitivity of insulin resistance of HepG2 cells to insulin.

CONCLUSIONHigh concentration of ROS can promote the proliferation of HepG2, and however low concentration of ROS inhibits the proliferation of HepG2. ROS can significantly improve insulin resistance of HepG2 cells induced by high insulin.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Glucose ; metabolism ; pharmacology ; Humans ; Hypoglycemic Agents ; isolation & purification ; pharmacology ; Insulin ; pharmacology ; Insulin Resistance ; Liver Neoplasms ; metabolism ; pathology ; Oligosaccharides ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rehmannia ; chemistry

OBJECTIVETo evaluate the feasibility and safety of nickel-titanium compression anastomosis ring (CAR27) in colorectal anastomosis after low anterior rectal resection in animal models.

METHODSEnd-to-end colorectal anastomosis was performed using CAR27 in 6 experimental pigs after resection of the middle and lower third of the rectum. The animals were observed postoperatively for up to 56 days. Five pigs were sacrificed at day 14 and the other at day 56. Distance from anal verge to anastomosis and anastomotic circumference were measured. Histopathologic examination was performed.

RESULTSThe median distance from anal verge was 5.3(4-6) cm. No anastomotic leak or other complications were observed. All the pigs recovered and gained weight. In 5 animals sacrificed at day 14, the mean circumference of the anastomosis was 6.8(6.5-7.0) cm, and histopathological examination showed mild inflammatory reaction and fibrosis. In the one sacrificed at day 56, the circumference expanded to 9.3 cm, and no inflammation and fibrosis were observed. Minor adhesion was noticed in only one pig, while smooth and intact serosa in the anastomosis was seen in the rest of the animals.

CONCLUSIONCAR27 is a promising device for mid and low colorectal anastomosis.

Anastomosis, Surgical ; instrumentation ; Animals ; Female ; Male ; Models, Animal ; Nickel ; Rectal Neoplasms ; surgery ; Rectum ; surgery ; Swine ; Swine, Miniature ; Titanium

OBJECTIVETo investigate the distribution of pathogenic C.albican genotype and Candida species in association with the severity of vulvovaginal candidiasis (VVC).

METHODSPolymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) of the internal transcribed spacer analysis was employed to identify the Candida species isolated from the vaginal secretions of 198 patients with acute VVC. SSCP and GeneScan analyses of microsatellite locus I polymorphism were used to determine the genotypes of the clinical isolates of C. albican associated with VVC. All the patients were scored for clinical signs and symptoms to evaluate the severity of VVC.

RESULTSA total of 198 Candida strains were isolated from VVC patients, including 140 (70.7%) C. albicans strains and 58 (29.3%) non-albicans strains. In the 95 patients with severe VVC and 103 with mild-moderate VVC, C.albican was detected in 62.1% and 76.6% of the patients, respectively (P=0.011). Thirty-eight microsatellite locus I genotypes were detected in 140 unrelated C. albican strains, among which the dominant genotypes 30-45 (44 strians, 31.43%) and 32-46 (23 strains, 16.43%) were the most common, followed by genotypes 30-46 (4 strains, 2.86%) and 32-47 (9 strains, 6.42%). The overall frequencies of the 4 genotypes were significantly higher in severe VVC than in mild-moderate VVC cases (77.9% vs 42.0%, P<0.001).

CONCLUSIONC. albicans remains the most common pathogenic Candia species in patients with VVC, but the non-alibcans species seem more likely to cause severe VVC. The dominant genotypes of C. albicans with a tropism for the vagina are correlated to the severity of VVC.

Adolescent ; Adult ; Candida ; classification ; isolation & purification ; Candida albicans ; genetics ; isolation & purification ; Candidiasis, Vulvovaginal ; microbiology ; Female ; Genotype ; Humans ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Single-Stranded Conformational ; Severity of Illness Index ; Young Adult

OBJECTIVETo investigate the effects of total flavonoid extract of Epimedium sagittatum (TFE) on the proliferation and differentiation of newborn rat calvarial osteoblasts (ROB).

METHODTFE was supplemented into the culture medium of ROB at 0. 1, 1, 10 and 100 microg x mL(-1) respectively. The serum of rats administered TFES (SRAT) was also added into the medium in a parallel treatment at 2.5%, 5% and 10% respectively. Their effects on cell proliferation and differentiation was studied by MTT and the analysis of osteogenic differentiation marks.

RESULTTFE had no appreciable and on cell proliferation and differentiation at any concentration. However, 2.5% and 5% SRAT stimulated cell proliferation strongly and, 5% SRAT significantly promoted the maturation and function of osteoblast by improving the alkaline phosphatase activity, osteocalcin secretion, calcium deposition and the number of mineralized nodular structures.

CONCLUSIONThe metabolites of TFE should be the anti-osteoporosis constitutes of Epimedium sagittatum.

Animals ; Calcification, Physiologic ; drug effects ; Calcium ; metabolism ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Flavonoids ; blood ; pharmacology ; Osteoblasts ; cytology ; drug effects ; metabolism ; Rats ; Rats, Wistar

Objective:To evaluate the effect of high-concentration hydrogen inhalation on sepsis-associated encephalopathy (SAE) in mice.Methods:Healthy male ICR mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=50 each) using the random number table method: sham operation group (Sham group), SAE group, sham operation plus high-concentration hydrogen group (Sham+ H 2 group), and SAE plus high-concentration hydrogen group (SAE+ H 2 group). SAE model was prepared by cecal ligation and puncture (CLP) in anesthetized animals.At 1 and 6 h after operation, Sham+ H 2 and SAE+ H 2 groups inhaled the mixture of hydrogen and oxygen (67% hydrogen-33% oxygen) for 1 h, and Sham and SAE groups inhaled the mixture of nitrogen and oxygen (67% nitrogen-33% oxygen) for 1 h. The postoperative 7-day survival rate was recorded.Cognitive function was assessed by Y maze test at days 3, 5 and 7 after operation.The mice were sacrificed at 24 h after operation, and hippocampal tissues were obtained for microscopic examination of the pathological changes of neurons in hippocampal CA1 region (with a light microscope) and for determination of normal neuron count, contents of tumor necrosis factor-ɑ (TNF-α) and high mobility group box-1 (HMGB1) (by enzyme-linked immunosorbent assay), mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry) and content of mitochondrial ATP (by fluorescein-fluorescent enzyme luminescence method). Results:Compared with Sham group, the 7-day survival rate after operation, percentage of spontaneous alternation at each time point after operation, and the number of normal neurons were significantly decreased, the contents of TNF-ɑ and HMGB1 were increased, and the contents of ATP and MMP were decreased in SAE and SAE+ H 2 groups ( P<0.05), and no significant change was found in Sham+ H 2 group ( P>0.05). Compared with SAE group, the 7-day survival rate after operation, percentage of spontaneous alternation at each time point after operation, and the number of normal neurons were significantly increased, the contents of TNF-ɑ and HMGB1 were decreased, and the contents of ATP and MMP were increased in SAE+ H 2 group ( P<0.05). Conclusions:High-concentration hydrogen inhalation can reduce SAE, and the mechanism may be related to reduction of hippocampal inflammatory responses and improvement in mitochondrial function in mice.