A method based on gas chromatography-mass spectrometry (GC-MS) was established to analyze the changes of intracellular metabolites and study the toxic mechanisms of different concentrations of particulate matter (PM2.5) effecting the lung tissues in mice.Nasal drip experiments of PM2.5 suspensions (0, 7.5, 20.0, 37.5 g/L) for mice were carried out, and the intracellular metabolites in lung tissues were extracted, pretreated and analyzed.Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were employed for pattern recognition, and an obvious distinction among different conditions was found.According to the PLS-DA loading diagram and variable important factor (VIP) value, 7 kinds of potential biomarkers, alanine, valine, leucine, ornithine, fumaric acid, citric acid and purine (p<0.01), were determined with significant differences between four different concentrations of PM2.5.Metabolic pathway analysis indicated that the oxidative stress reactions were enhanced, and the TCA cycle and the purine metabolism in lung cells were restrained after dripping PM2.5 to the lung tissues in mice.This study could provide a new perspective and theoretical basis for the further analysis on toxic mechanisms by PM2.5.

[Summary] Type 2 diabetic rat model accompanied by insulin resistance was established by high fat/high glucose diet and streptozotocin.Following metformin treatment for 4 weeks,realtime PCR and Western blot were used to detect the expressions of SIRT3 mRNA and protein in skeletal muscle tissue,respectively.The results showed that insulin sensitivity,superoxide dismutase (SOD),and glutathione (GSH) levels were significantly reduced in diabetic group compared with the control group (P＜0.05),while methane dicarboxylic aldehyde (MDA) level was increased (P＜0.05),along with the decreased expressions of SIRT3 mRNA and protein in skeletal muscles (P＜0.01).After metformin treatment,the insulin sensitivity,SOD,GSH,and SIRT3 mRNA and protein levels were significantly increased (P＜ 0.05 or P＜ 0.01),while MDA level was decreased (P ＜ 0.05),suggesting that metformin may ameliorate insulin resistance via upregulating SIRT3 expression in skeletal muscles of diabetic rats.

Objective To observe the physique and anatomy changes in patients with nasopharyngeal carcinoma (NPC) during intensity-modulated radiotherapy (IMRT), using repeated CT images and deformable registration technique, and analyze their impact on delivery dose distribution.Methods Ten NPC patients were randomly selected from those who had received IMRT treatment.Gross tumor volume of nasopharyn (GTVnx), GTV of metastastatic lymph node (GTVnd), clinical target volume (CTV) and normal tissue or organ (OAR) were re-contoured on the in-course repeated CT images using a kind of deformable registration and auto-segmentation software according to the original planning contouring.Changes in volume of treatment targets and organs at risk were evaluated and the trends were then analyzed.Dose distributions were recalculated with repeated CT images and compared to the original plans.Results The volume of GTVnx were decreased by 6.44%,10.23% and9.72%(F=1.34,P=0.278) in the 2-,4-and 6-week after IM RT comparing with before IM RT, with 6.59%, 30.98 % and 35.13 % (F = 9.22, P =0.000) in GTVnd, 0.73%, 1.86% and 1.41% (F=0.33,P=0.722) in CTV1, -1.78%, -6.47%and -9.34% (F =16.89 ,P =0.000) in CTV2, 13.96%, 32.97% and 37.77%(F=17.17,P=0.000)in the left parotid , and 3.56% , 29.57% and 35.63% (F = 13.49 , P = 0.000) in the right parotid.The mean dose change rate of GTVnx were -0.39%, 0.08% and 0.32% (F =0.15 ,P =0.860) in the 2-,4- and 6-week after IMRT comparing with planning faction dose, with 0.53%, 1.19% and 0.69% (F=0.81,P=0.455) in GTVnd, 1.95%, 2.70% and 3.78% (F=0.61,P=0.552) in the spinal cord,0.32%, 0.81% and 0.62% (F=0.03,P=0.975) in the brain stem, 4.50%, 4.66% and 7.20% (F=0.33,P=0.725) in the left parotid, 2.20%, 7.17% and 7.12% (F= 1.24,P=0.306) in the right parotid.Conclusions The GTVnd, CTV2 and parotids shrinks obviously along with the treatment times for NPC patients during IMRT.Although changes in fraction dose of GTV, CTV, spinal cord, stem and parotids are not significant, further study with larger samples is needed.

To explore the expression potential of heterogeneous genes using the backbone of infectious bronchitis virus (IBV) Beaudette strain, the ectodomain region of the Spike gene (1,302 bp) of IBV H120 strain was amplified by RT-PCR and replaced into the corresponding location of the IBV Beaudette strain full-length cDNA. This recombinant was designated as BeauR-H120(S1). BeauR-H120(S1) was directly used as the DNA template for the transcription of viral genomic RNA in vitro. Then, the transcription product was transfected into Vero cells by electroporation. At 48 h post-transfection, the transfected Vero cells were harvested, and passaging continued. A syncytium was not observed until the recombinant virus had passed through four passages. The presence of rBeau-H120(S1) was verified by the detection of the replaced ectodomain region of the H120 Spike gene using RT-PCR. Western blot analysis of rBeau-H120 (S1)-infected Vero cell lysates demonstrated that the nucleocapsid (N) protein was expressed, which implied that rBeau-H120(S1) could propagate in Vero cells. The TCIDs0 and EIDs0 data demonstrated that the titer levels of rBeau-H120(S1) reached 10(590+/-0.22)TCID50/mL and 10(6.13+/-0.23)EID50/mL in Vero cells and 9-day-old SPF chicken embryos, respectively. Protection studies showed that the percentage of antibody-positive chickens, which were vaccinated with rBeau-H120(S1) at 7 days after hatching, rose to 90% at 21 days post-inoculation. Inoculation provided an 85% rate of immune protection against a challenge of the virulent IBV M41 strain (103EID50/chicken). This recombinant virus constructed using reverse genetic techniques could be further developed as a novel genetic engineering vaccine against infectious bronchitis.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Coronavirus Infections ; veterinary ; virology ; Infectious bronchitis virus ; chemistry ; genetics ; growth & development ; metabolism ; Poultry Diseases ; virology ; Protein Structure, Tertiary ; Spike Glycoprotein, Coronavirus ; chemistry ; genetics ; metabolism ; Transfection ; Vero Cells

OBJECTIVETo investigate the effects of the furin inhibitor α1-PDX on the growth, invasion, and tumorigenicity of cervical cancer cells and explore the mechanisms.

METHODSThe changes in the growth, migration and invasion of α1-PDX-transfected HeLa cells were observed using MTT assay, Boyden migration and invasion assay. The protein levels of furin and MT1-MMP were measured using Western blotting and furin activity was detected by enzyme activity assay in the transfected cells. HeLa cells were seeded subcutaneously in nude mice and the tumor volume changes were recorded.

RESULTSCompared with the control cells, α1-PDX-treated cells showed a significant growth inhibition by 18.4% at 24 h (P<0.01) with obviously lowered migration ability and cell invasiveness (P<0.01). Treatment with α1-PDX significantly reduced furin enzyme activity and MTI-MMP protein levels in HeLa cells. In nude mice, α1-PDX-treated HeLa cells exhibited a delayed and lowered tumorigenicity with reduced size of the tumors.

CONCLUSIONα1-PDX can inhibit the growth, metastasis and tumorigenicity of HeLa cells, the mechanism of which may involve a decreased furin activity and MTI-MMP expression.

Animals ; Female ; Furin ; antagonists & inhibitors ; HeLa Cells ; drug effects ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Transfection ; Uterine Cervical Neoplasms ; pathology ; alpha 1-Antitrypsin ; pharmacology

Objective To evaluate the relationship between the failed mechanism of sevoflurane postconditioning-induced myocardial protection and the activity of dynamin-related protein 1(Drp1)in dia-betic rats. Methods Pathogen-free healthy adult male Sprague-Dawley rats, weighing 220-280 g, in which diabetes mellitus was induced by combination of high-fat and high-sucrose diet and intraperitoneal injection of streptozotoein 30 mg∕kg, were studied.Sixty rats with diabetes mellitus were divided into 5 groups(n=12 each)using a random number table: sham operation group(group Sham), myocardial ischemia∕reperfusion (I∕R)group(group I∕R), sevoflurane postconditioning group(group SP), Drp1 inhibitor mitochondrial division inhibitor-1(Mdivi-1)group(group M)and Mdivi-1 plus sevoflurane postconditioning group(group M-SP). Myocardial I∕R was induced by occluding the left anterior descending branch of the coronary artery for 30 min followed by 120 min reperfusion except for group Sham. Mdivi-1 1.2 mg∕kg was intraperito-neally injected at 15 min before ischemia in M and M-SP groups, and 2.5% sevoflurane was inhaled starting from 5 min of reperfusion in SP and M-SP groups. Blood samples were collected from the right internal jugular vein at 120 min of reperfusion for measurement of serum cardiac troponin I(cTnI)concentrations(by en-zyme-linked immunosorbent assay). Rats were then sacrificed and myocardial specimens were obtained for de-termination of the myocardial infarct size(by TTC), cell apoptosis(by TUNEL), expression of Bax, Bcl-2 and activated caspase-3(by Western blot)and nicotinamide adenine dinucleotide(NAD+)content(by spectrophotometry). Apoptosis index(AI)and Bax∕Bcl-2 ratio were calculated. Results Compared with group Sham, the percentage of myocardial infarct size, serum concentration of cTnI, AI and Bax∕Bcl-2 ratio were significantly increased, the expression of activated caspase-3 was up-regulated, and the NAD+content was decreased in the other four groups(P<0.05). Compared with group I∕R, the percentage of myocardial infarct size, serum concentration of cTnI, AI and Bax∕Bcl-2 ratio were significantly decreased, the expres-sion of activated caspase-3 was down-regulated, and the NAD+content was increased in group M-SP(P<0.05), and no significant change was found in the parameters mentioned above in SP and M groups(P>0.05). Compared with group SP, the percentage of myocardial infarct size, serum concentration of cTnI, AI and Bax∕Bcl-2 ratio were significantly decreased, the expression of activated caspase-3 was down-regulated, and the NAD+content was decreased in group M-SP(P<0.05), and no significant change was found in the parameters mentioned above in group M(P>0.05). ConclusionThe failed mechanism of sevoflurane postconditioning-induced myocardial protection may be related to the activity of Drp1 in diabetic rats.

Objective To investigate the effects of the furin inhibitor α1-PDX on the growth, invasion, and tumorigenicity of cervical cancer cells and explore the mechanisms. Methods The changes in the growth, migration and invasion of α1-PDX-transfected HeLa cells were observed using MTT assay, Boyden migration and invasion assay. The protein levels of furin and MT1-MMP were measured using Western blotting and furin activity was detected by enzyme activity assay in the transfected cells. HeLa cells were seeded subcutaneously in nude mice and the tumor volume changes were recorded. Results Compared with the control cells, α1-PDX-treated cells showed a significant growth inhibition by 18.4% at 24 h (P<0.01) with obviously lowered migration ability and cell invasiveness (P<0.01). Treatment withα1-PDX significantly reduced furin enzyme activity and MTI-MMP protein levels in HeLa cells. In nude mice, α1-PDX-treated HeLa cells exhibited a delayed and lowered tumorigenicity with reduced size of the tumors. Conclusionα1-PDX can inhibit the growth, metastasis and tumorigenicity of HeLa cells, the mechanism of which may involve a decreased furin activity and MTI-MMP expression.

Objective To investigate the effects of the furin inhibitor α1-PDX on the growth, invasion, and tumorigenicity of cervical cancer cells and explore the mechanisms. Methods The changes in the growth, migration and invasion of α1-PDX-transfected HeLa cells were observed using MTT assay, Boyden migration and invasion assay. The protein levels of furin and MT1-MMP were measured using Western blotting and furin activity was detected by enzyme activity assay in the transfected cells. HeLa cells were seeded subcutaneously in nude mice and the tumor volume changes were recorded. Results Compared with the control cells, α1-PDX-treated cells showed a significant growth inhibition by 18.4% at 24 h (P<0.01) with obviously lowered migration ability and cell invasiveness (P<0.01). Treatment withα1-PDX significantly reduced furin enzyme activity and MTI-MMP protein levels in HeLa cells. In nude mice, α1-PDX-treated HeLa cells exhibited a delayed and lowered tumorigenicity with reduced size of the tumors. Conclusionα1-PDX can inhibit the growth, metastasis and tumorigenicity of HeLa cells, the mechanism of which may involve a decreased furin activity and MTI-MMP expression.

BACKGROUND: Complex regional pain syndrome (CRPS) is a painful, disabling disorder for which no proven treatment has been established. The purpose of this investigation was to assess the evidence of the efficacy of spinal cord stimulation (SCS) in the management of pain in CRPS patients. METHODS: Between March 2004 and June 2006, 11 patients with CRPS were treated with SCS. The visual analog scale (VAS) score for pain (0-10) and pain disability index (PDI) were obtained in all patients prior to treatment, and 1, 3 and 6 months post-implantation. RESULTS: All 11 patients, 5 men and 6 women, with a median age and duration of CRPS of 44 years and 48.8 months, respectively, successfully received a lead implantation for SCS. The mean VAS pain score prior to the treatment was 85.5 out of 100 mm. After SCS implantation, the mean VAS pain scores were 49.5, 57.0 and 56.0 at 1, 3 and 6 months after the procedure, respectively. The mean pain score for allodynia was decreased by 50%, with a significant reduction of the PDI also observed after the treatment. CONCLUSIONS: Our current study suggests that SCS implantation is a safe and effective method in the management of CRPS patients.
Female ; Humans ; Hyperalgesia ; Male ; Spinal Cord Stimulation* ; Spinal Cord* ; Visual Analog Scale

Objective: To explore the difference of liver inflammation and fibrosis in patients with chronic hepatitis B virus (HBV) infection and chronic hepatitis C virus (HCV) infection, and to investigate the relationship between hepatic pathology and alanine aminotransferase (ALT). Methods: 57 patients with chronic HCV infection and 346 patients with chronic HBV infection who were hospitalized at Shengjing Hospital of China Medical University from January 2012 to September 2016 were enrolled. In chronic HBV infection, including 88 cases whose ALT were more than two times of upper limited of normal (ALT≥2×ULN) and 258 cases whose ALT were less than two times of upper limited of normal (ALT < 2×ULN).All the patients were underwent liver biopsy. Chronic HBV infection (ALT≥2×ULN and ALT < 2×ULN) and chronic HCV infection were compared respectively. Statistical analyses were performed using a Univariate χ²-test and Mann–Whitney U test for comparison. Correlations between variables were analyzed using Spearman's rank correlation. Results: In chronic HBV infection group, 169 cases (48.8%) had inflammation grade≥2 (G≥2), 98 cases (28.3%) had fibrosis stage≥2 (S≥2), 81 cases (23.4%) with G≥2 and S≥2.In the ALT < 2×ULN group, there were 109 cases (42.2%) with G≥2, 62 cases (24%) with S≥2, 49 cases (19%) with G≥2 and S≥2. In the ALT≥2×ULN group, 60 cases (68.2%) with G≥2, 35 cases (39.8%) with S≥2, 31 cases (35.2%) with G≥2 and S≥2. The grade of inflammation and fibrosis have significantly different between ALT≥2×ULN group and ALT < 2×ULN group (χ² = 17.66, χ² = 8.06, P < 0.01). In chronic HCV infection group, 47 cases (82.5%) with G≥2, 20 cases (35.1%) with S≥2, 20 cases (35.1%) with G≥2 and S≥2. ALT had no correlation with inflammation and fibrosis (P > 0.05). The grade of inflammation was significantly different between chronic HCV infection and chronic HBV infection whose ALT < 2×ULN (χ² = 30.19, P < 0.01) but the fibrosis have no difference (χ² = 2.96, P > 0.05). Compared with chronic HBV infection whose ALT≥2×ULN, both inflammation and fibrosis had no significantly different (χ² = 3.65, χ² = 0.32, P > 0.05 respectively). Conclusion In chronic HBV infection whose ALT < 2×ULN, about 30%-40% liver tissue with significant necroinflammation and /or fibrosis. About 80% chronic HCV infection with significant necroinflammation, and the grade of inflammation has no correlation with ALT. The grade of inflammation has significantly different between chronic HCV infection group and chronic HBV infection group whose ALT < 2×ULN.