ObjectiveTo investigate the mechanism of protective effect of Xuebijing injection on vascular endothelial cells in rats with heat stress.Methods Ninety Sprague-Dawley(SD) rats were randomly divided into control, model and Xuebijing injection treatment groups, 30 rats in each group. Heat stress model was reproduced by placing rats in constant temperature box at 40℃, 60% relative humidity for 1 hour, Xuebijing injection group was treated by intraperitoneal injection of Xuebijing 2.5 g/kg, while the control and model groups were treated by intraperitoneal injection of normal saline 2 mL/kg, once a day only in 1 day for both groups. After model establishment, the rectum temperature, heart rate and the mean arterial pressure(MAP) were recorded at 2, 6, 12 hours in each group. At the same time, the rat abdominal aortic blood was collected and serum was separated, the enzyme-linked immunosorbent assay(ELISA) was used to determine the aortic serum levels of lipopolysaccharide(LPS), nuclear factor-κB(NF-κB) and p53, and the prothrombin time(PT), activated partial thromboplastin time(APTT) and D-dimer of venous blood were detected by automatic blood coagulation analyzer(ACLTOP).Results Compared with those in control group, the rectum temperature, heart rate, LPS, NF-κB, p53, PT, APTT, D-dimer were significantly increased, and MAP was obviously decreased in model group(P<0.05 orP<0.01). Compared with model group, the above indexes were improved significantly in Xuebijing injection treatment group at 2 hours〔rectum temperature(℃): 38.02±0.22 vs. 39.32±0.33, heart rate(bpm): 507±14 vs. 562±35, MAP(mmHg, 1 mmHg=0.133 kPa): 98±6 vs. 87±13, LPS(ng/L): 0.65±0.03 vs. 0.82±0.05, NF-κB(ng/L): 1.10±0.04 vs. 1.33±0.05, p53(ng/L): 1.33±0.03 vs. 1.73±0.02, PT(s):15.47±1.03 vs. 20.28±2.01, APTT(s): 40.26±2.46 vs. 47.46±3.51, D-dimer(μg/L): 238.54±8.32 vs. 323.12±8.14,P<0.05 orP<0.01〕.Conclusion Xuebijing injection can correct the disorders of blood PT, APTT, D-dimer via decreasing the secretion of the levels of NF-κB, p53 from vascular endothelial cells in rats with heat stress, thus the integrity of the vascular endothelium can be protected, and LPS entering into the blood stream can be inhibited.

Objective To explore the effectiveness of ω-3 fatty acids in intervening rat models of chronic obstructive pulmonery disease (COPD).Methods The rat COPD models were established by cigarette smoking and intratracheal lipopolysaccharide instillation.Totally 90 rats were randomly divided into 3 groups:normal control group (treated with normal saline),COPD group,and intervention group (the COPD rat models treated with ω-3 fatty acids).Lung tissues were obtained on the 7th,21st,and 28th day.The left lower lobes were analyzed to determine the expressions of nuclear factor-kappa beta (NF-κB) and interferon-γ (IFN-γ)and the right lung lobes were sliced for detecting the cell apoptosis.Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was used to detect the serum IFN-γ and interleukin-6 (IL-6).Results (1) The pathological changes of lung tissue:there were a large number of inflammatory exudation,alveolar wall thickening,hyperplasia of vascular smooth muscle and the alveolar structure destruction in the COPD model group,but the lung tissue were part of alveolar cavity and a little inflammatory exudate in ω-3 fatty fish acids treatment group,control rats were almost no alveolar inflammation on the 28th days.(2) On the 28th day,NF-κB protein expression of the lung tissue (18.91 ± 3.07) in rats of COPD model group was significantly higher than the control group and the intervention group (5.47 ±4.86 and 7.23 ±2.21) (P ＜0.01).On the 28th day,IFN-γ protein expression in lung tissue of the rats in the model group was 7.12 ±3.37,significantly lower than the intervention group (18.74 ± 2.65),the difference was statistically significant (P ＜ 0.01).(3) The IL-6 levels of the blood-serum of model group rats were (13.43 ± 2.47) ng/L,significantly higher than the control group and the intervention group [(4.78 ± 1.93) and (4.98 ± 1.89) ng/L],the differences were statistically significant (P ＜ 0.01) on the 28th day,,but the IFN-γ level [(2.23 ± 0.63) ng/L] in COPD group was more poorer than ω-3 fatty acids group and the intervention group [(4.51 ± 0.71) and (7.05 ± 0.52) ng/L] (P ＜ 0.01).Conclusions The ω-3 fatty acids can lower NF-κB protein expressions in lung tissues and serum and IL-6 levels in COPD rats; aslo,it can increase the IFN-γ protein expression in lung tissue and serum.Thus,it can prevent the lung inflammation in COPD rats.

Objective To investigate the age-associated changes of ultrastructure,mRNA and protein expressions of H+-K+-ATPase in elderly gastric parietal cell. Methods Fifty patients with relative normal stomach without gastroduodenal diseases were enrolled,including younger group (aged 20-59 years,n=19) and elderly group (aged≥60 years,n=31).Furthermore,the elderly group was divided into 3 subgroups:60-69 years old (n =11 ),70-79 years old (n=10 ),above 80 years old (n =10).The ultrastructure of gastric parietal cell was observed under electron microscope.The expression of H+-K+-ATPase α subunit mRNA and H+-K+-ATPase β subunit protein were assessed by quantitative real-time PCR and Western-blot,respectively.The ageing-associated changes of all these data were respectively compared. Results No significant difference was showed in the morphology of gastric parietal cell and acid-secretion-associated organelles among all the groups.The average ratio Am to Ac (Am means the area of mitochondria,Ac means the area of cytoplasm) of gastric parietal cell and the average At to Ac ratio (At means the area of secretory canaliculi and tubulovesicular system )between younger group and elderly group had no significant difference[(48.4±7.5) ％ vs.(50.6±7.6) ％,t=-0.775,P=0.444; (13.8±4.1) ％ vs.(12.2±4.7) ％,t=0.984,P=0.332].Meanwhile,there were no distinctions in the expression of H+-K+ -ATPase α subunit mRNA and H+-K+-ATPase protein among all elderly subgroups(F=1.522,2.32,P=0.24,0.114).However,the mRNA expression of H+-K+-ATPase a subunit was higher in the elderly group than in the younger group(t=-3.682,P=0.001).Furthermore,the expression of H+ -K+ -ATPase protein in the elderly group was increased as compared with younger group(t=-3.389,P=0.004). Conclusions Acidsecretion-associated organelles of human gastric parietal cell have no degeneration and the expression of H + -K+-ATPase is in trend of increase with aging,indicating that healthy elderly people have the basis of ultrastructure and molecular biology to maintain well function of acid secretion.

Animals ; Guinea Pigs ; Heart Ventricles ; drug effects ; physiopathology ; Male ; Stilbenes ; pharmacology

Objective The detection of malignant lymphoma with invasion in liver and spleen using PET/CT has not been well documented in the literature. This study aimed to investigate the usefulness of PET/CT in this regard and to compare it with plain CT. Methods Forty-one pathologically confirmed malignant lymphoma patients with liver and spleen invasion were recruited into this study. Among all patients, there were 38 non-Hodgkin's lymphoma (NHL), 2 Hodgkin's lymphoma (HL) and 1 gastric mucosa associated lymphoma. PET/CT imaging was recorded 1h after injection of 296～444 MBq 18F-fluorodeoxyglucose (FDG). Results (1) There were 30(30/41) patients with liver invasion, including hepatic nodules, mass and portal nodes. The mass was large to invade surrounding liver parenchyma. (2) There were 23(23/41) patients with spleen invasion. The spleen was enlarged and demonstrated diffused hyper-metabolism. (3) Other invasion included: lung (n=13), cortical bone and marrow (n=12), stomach (n=9), pleural (n=6), and subcutaneous soft tissue (n=5) and so on. Conclusion PET/CT could accurately diagnose the invasion in liver and spleen of malignant lymphoma, which was of potential role on the diagnosis and staging of lymphoma.

Objective To investigate the titer stability of the harvest solution of severe acute respiratory syndrome coronavirus2(SARS-CoV-2)at 2 ～ 8 ℃ and the inactivation effect of β-propiolactone inactivator on the virus.Methods Three batches of SARS-CoV-2 harvest solution(batch numbers:202111001,202111002 and 202111003)were stored at 2 ～ 8 ℃ for 12 d and sampled every 3 d(0,3,6,9 and 12 d)for detection of the titers by Karber method;Three batches of virus harvest solution equilibrated overnight at 2 ～ 8 ℃ were inactivated by adding β-propiolactone at a volume fraction of 1∶4 000 and detected for the titers at different inactivation time points(0,0.5,1,1.5,2,3,4,8,16 and 24 h),of which samples inactivated for 8,16 and 24 h were taken for inactivation verification,and samples inactivated for 24 h were observed by transmission electron microscope.Results The titers of SARS-CoV-2 decreased with the prolongation of storage time at 2 ～8 ℃,which showed no obvious decrease during 0 ～ 3 d,while decreased from the initial 7.75,6 and 7.5 lgCCID_(50)/mL to5.75,4.625 and 6.25 lgCCID_(50)/mL on day 12,indicating that the virus activity showed a gradual decrease trend at 2 ～8 ℃；With the inactivation time,the virus titer decreased continuously and could not be detected after inactivation for 3 h.Transmission electron microscope observation showed that the inactivated virus particles were intact and the spike protein was evenly distributed.Conclusion The virulence of SARS-CoV-2 stored at 2 ～ 8 ℃ was unstable,so the subsequent inactivation and purification process should be carried out as soon as possible;The titer of virus could not be detected after3 h of inactivation,which provided a reference for the determination of the inactivation process.

Objective To discuss the molecular mechanism of 18F-fluorodeoxyglucose (FDG) uptake in tumor and to assess its value to identify pathologic type and cancer staging in patients with earlystage nasopharyngeal carcinoma.Methods Forty patients with nasopharyngeal carcinoma of early-stage,including 12 cases with T1 stage and 28 cases with T2 stage, underwent FDG PET imaging.The maximum standardized uptake value ( SUVmax ) and mean standardized uptake value ( SUVmean ) of FDG uptake of each patient were measured and compared between T1 and T2 stage by t-test.The expression of glucose transport protein 1 ( Glut1 ) and hexokinase- Ⅱ ( HK- Ⅱ ) of each case was measured in paraffin sections by streptavidin-perosidase (SP) immunohistochemistry.The positive expression rate of Glut1 and HK- Ⅱ was calculated and compared between T1 and T2 by x2 test.Meanwhile, the correlation between the expression of Glut1 or HK-Ⅱ and the SUVmax was tested by Pearson analysis.Results The SUVmax and SUVmean in 40 patients were 9.45 ± 1.87 and 6.04 ± 1.09, respectively.The SUVmax of patients with T1 stage (8.95 ± 1.91 ) was significantly lower (t =4.46, P＜0.001 ) than that of patients with T2 stage (11.55 ± 1.70), and the SUVmean of patients with T1 stage (5.61 ± 1.08) was significantly lower ( t = 6.76, P ＜ 0.001 ) than that of patients with T2 stage (7.98 ± 1.10) too.Among 40 patients, all patients showed positive expression of Glut1 and HK-Ⅱ , and the positive expression rate of Glut1 and HK-Ⅱ was ( 45.2 ± 10.9 )% and ( 68.3 ±9.5)%, respectively.The positive expression rate of Glut1 was (38.4 ±8.1)% in T1 stage and (49.7 ±12.6)% in T2 stage, which displayed no difference (x2 =40.58, P＞0.05), but the HK-Ⅱ positive expression rate showed significant difference (x2 =58.71, P＜0.05) between T1 stage (60.1 ±11.1)% and T2 stage (77.9 ± 14.7 )%.The correlation analysis indicated that there was low-degree positive correlation (r =0.369, P=0.019) between the SUVmax and Glut1 expression, and there was medium-degree positive correlation (r = 0.549, P = 0.001 ) between the SUVmax and HK-Ⅱ expression.Conclusion Expression of Glut1 and HK-Ⅱ was positively correlated with FDG uptake in patients with early-stage nasopharyngeal carcinoma.

OBJECTIVETo explore the relation of the standard uptake values (SUV) of 18F-fluorodexygl-gucose (18F-FDG) PET/CT and the pathological classification and clinical staging of nasopharyngeal carcinoma (NPC).

METHODWhole body 18F-FDG PET imaging was performed in 32 patients with pathologically confirmed NPC, who received no previous treatment. The regions of interest (ROI) covering the pharyngeal and cervical lesions were defined along the margins of the lesion, and the lesion volume and the SUVs were calculated.

RESULTSThe SUVs in stage I, II, III, and IV patients were 4.50-/+0.42, 5.62-/+1.44, 7.33-/+1.50, and 8.24-/+2.16, respectively, showing significant differences between them (P < 0.05). In patients in stage T1, T2, T3, and T4, the SUVs increased significantly in advanced stageds (2.56-/+1.05, 3.72-/+0.60, 6.87-/+1.07, and 9.70-/+0.70, respectively, P < 0.05). The SUVs were not significantly different in patients in stages N1, N2, and N3, differed significantly between lymph nodes > 6 cm in size and those < or = 6 cm (5.92-/+1.51 vs 3.48-/+1.31, P < 0.05). The SUV in poorly differentiated squamous carcinoma was significantly lower than that in undifferentiated carcinoma (5.58-/+1.48 vs 8.41-/+1.71, F = 1.3323, P = 0.01).

CONCLUSIONSThe SUV is not associated with the clinical staging of NPC, but is correlated to the T staging of NPC. Though irrelevant to the N staging of NPC, the SUV is correlated to the size of the lymph nodes, and also related to the degree of differentiation of NPC.

Adolescent ; Adult ; Female ; Fluorodeoxyglucose F18 ; pharmacokinetics ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnostic imaging ; pathology ; Neoplasm Staging ; Positron-Emission Tomography ; ROC Curve ; Tomography, X-Ray Computed

OBJECTIVETo explore the effect of berberine on lipid metabolism disorder and lipid deposition in liver cells of non-alcoholic fatty liver disease (NAFLD) rats induced by high fat diet.

METHODSAfter one week adaptable feeding, 45 SPF level male SD rats were randomly divided into 3 groups, the normal control group, the model group, and the berberine group, 15 in each group. Except those in the normal control group, all rats were fed with high fat diet to prepare NAFLD model. As for rats in the berberine group, Berberine Hydrochloride was administered by gastrogavage. HE staining and oil red O staining were performed to identify the model after 8 weeks. Hepatocytes were isolated, and their activities and purities were tested by Typan blue staining and flow cytometry (FCM). Serum levels of TC, TG, HDL-C, and LDL-C were detected using automatic biochemical analyzer. mRNA expression levels of LXRα and FAS in liver cells were analyzed by Real-time quantitative polymerase chain reaction (PCR). Protein levels of LXRα and FAS in liver cells were examined by Western blot.

RESULTSThe NAFLD rat model was successfully established by high fat diet. The yields of purified liver cells in each rat were (6.0-7.5) x 10(8). The viability of isolated liver cells with purity over 90% (tested by FCM analysis) was higher than 95%. Compared with the normal control group,the expression of LXRα and FAS at mRNA and protein levels was higher in the model group (P < 0.01). Compared with the model group, the expression of LXRα and FAS at mRNA and protein levels was obviously down-regulated in the berberine group (P < 0.01).

CONCLUSIONSLXRα/FAS signaling pathway was one of important signaling pathways of NAFLD lipid metabolism disorders. Berberine could recover hepatocyte fatty deposits in NAFLD rats by adjusting the LXR/FAS signaling pathway of hepatocytes, which might be one of important mechanisms for fighting against NAFLD.

Animals ; Berberine ; therapeutic use ; Diet, High-Fat ; Down-Regulation ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Liver ; Hepatocytes ; Lipids ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism