Objective To observe how the ACC transmits nociceptive information and how it regulates spinal noceciption.Methods A total of 32 male SD rats were randomly divided into four groups:Sham group,CCI group,ACC group,and AP-5 group.After light-dark transition test, forced swimming test (FST),paw withdrawal mechanical threshold (PWMT),paw withdrawal ther-mal latency (PWTL)had been measured,the rats were finally anesthetized,then ACC and the spinal cord was rapidly removed,the expression of cAMP-response element binding protein (pCREB)and extracellular regulated protein kinases (pERK)were measured by Western blot.Results Compared with the sham group,the PWMT of the CCI rats were significantly decreased,rats spent less time in the light compartment and number of transition were decreased (P <0.01 ).The immobility time in FST were also significantly prolonged (P <0.01).After AP-5 injected in bilateral ACC 13 days after CCI operation,the PWMT of the CCI rats were significantly increased,rats spent more time in the light compartment and number of transition were increased (P <0.01).The immobility time in FST were also significantly shortened (P <0.01).Compared with sham group,the expression of pCREB, pERK increased significantly in ACC in CCI group (P <0.01).The expression of pCREB,pERK in spinal cord was increased in CCI group and decreased in AP-5 group (P <0.01 ).Conclusion ACC facilitate the spinal nociception in a descending mode.

Objective To investigate the effects of methylthioadenosine phosphorylase (MTAP) on invasion and migration in breast cancer cells.Methods Human breast cancer cell line MCF-7 cells were treated with MTAPtargeted siRNA to diminish MTAP mRNA.MCF-7 cells proliferation was evaluated by cell counting kit-8,the analysis of cells invasion and migration was performed using Transwell chamber.The expressions of MTAP and matrix metalloproteinase 1 (MMP1) in cell extracts were detected by Western blotting.The experimental divided into blank contrd group,negative control group,MTAP-siRNA experimental group.Results The MCF-7 cells growth was promoted after knockdown the MTAP mRNA.MATP-siRNA experimental group 450 nm absorbance values at 24h,48 hand72 h of the control group were (112.3±11.9)％,(144.4±8.4)％,(169.3±9.4)％ respectively.Cell invasion analysis by Transwell chamber showed 570 nm absorbance values were 0.49 ± 0.06 (control),0.45 ± 0.07 (negative control) and 0.87 ± 0.07 (MTAP-siRNA) respectively.Cell migration analysis by Transwell chamber showed 570 nm absorbance values were 0.46 ± 0.06 (control),0.49 ± 0.08 (negative control)and 0.75 ± 0.07 (MTAP-siRNA) respectively.The expression of MMP1 in MCF-7 cells was upregulated after knockdown the MTAP mRNA.Conclusion The knockdown of MTAP in MCF-7 cell can increase the cells invasion and migration,and this may involve the the MMP1.

Objective To observe pain behavior and the expression of Fos in the related brain re? gions,including Anterior Cingulate Cortex ( ACC) ,Periaqueductal Gray ( PAG) ,Rostral ventromedial nucle?us ( RVM) in chronic constrictive injury ( CCI) rats and to explore whether ACC modulate spinal nociceptive transmission through endogenous descending facilitatory system. Methods A total of 48 male SD rats were randomly divided into six groups:Naive group,Sham group ( just separated the left sciatic nerves without liga?tion) ,CCI group ( the left sciatic nerve was ligated) ,CCI+P group ( on the 14 th day after surgery,intraper?itoneal injection of 10 mg/kg paroxetine 45 min before behavior test) ,ACC?Sham group ( bilateral microin?jection of 0.9% NaCl in ACC,1μl/each side) and ACC?AP5 group ( bilateral microinjection of AP5 25 mM in ACC,1μl/each side on the 13th day after surgery) . On the 14th day after light?dark transition test,forced swimming test,paw?withdrawal mechanical threshold( PWMT) and paw?withdrawal thermal patency( PWTL) were performed,the rats were terminally anesthetized and ACC,PAG,RVM and the spinal cord was rapidly removed,and then the expression of c?fos was measured by immunohistochemistry. Results ( 1) Rats in CCI group demonstrated nociceptive hypersensitivity and depressive?like behaviors compared with Naive rats, while rats in CCI+P group and in ACC?AP5 group showed less nociceptive hypersensitivity and less depres?sive?like behaviors compared with CCI group. (2)Compared with Naive group,the number of c?fos positive neurons in the bilateral ACC (left,surgery side,4920.6±1053.7;right,2059.3±409.1),VIPAG (9074.8± 2320.3),RVM (6195.4±895.0) and bilateral spinal cord (left,15148.8±3080.2;right,6400.2±1558.4) was significantly enhanced in CCI group, especially in the left side. In contrast, the amount of fos labeled neurons declined in the bilateral ACC (left,2776.4±820.1;right,1120.5±141.4),VIPAG (4002.2± 1171.8),RVM (2938.9±910.3) and bilateral spinal cord (left,8742.0±1131.0;right,3933.1±858.9) in CCI+P group and also declined in the bilateral ACC (left,3623.1±667.4;right,696.5±164.8),VIPAG (5668.8±1403.3),RVM (3972.3±851.7) and bilateral spinal cord (left,10675.4±1725.3;right,3818.3± 1085.1) in ACC?AP5 group. Conclusion Endogenous descending facilitatory system may contribute to ACC modulating spinal nociceptive transmission.

Objective To explore the diagnostic value of MRI special use of breast and uhrasonography in axillary lymph node metastasis of early breast cancer.Methods Clinical data of 136 Ⅰ-Ⅲ A breast cancer patients accepted MRI examination before surgery had been retrospectively studied,analysing diagnostic value of MRI and ultrasonography in axillary lymph node metastasis of early breast cancer.Results The sensitivity,specificity,and accuracy obtained by MRI were 83.3％,88.6％ and 86.3％.And these data of ultrasonography were 73.1％,76.7％ and 75.0％.The sensitivity,specificity,and accuracy obtained MRI were better than that of ultrasonography.The sensitivity,specificity,and accuracy of ≥ 50 years old patients were 70.0％,77.8％ and 75.0％.And ＜ 50 years old patients were 85.7％,92.3％ and 88.9％.The sensitivity,specificity,and accuracy of ＜ 50 years old patients were better than ≥ 50 years old patients.Conclusions The MRI special use of breast have an important value in axillary lymph node metastasis of early breast cancer,especially to gounger than 50 years old patients.It can provide a scientific basis of the clinical accurate treatment for early breast cancer patients.

AIM: To investigate the effects of simvastation on homocysteine-induced endothelial dysfunction and inflammatory response and the underlying mechanisms of such effects. METHODS: MTT assay was used to detect cell viability, and DCFH-DA assay was used to examine the levels of reactive oxygen species (ROS). Furthermore, Western blotting was performed to detect protein expression and electrophoretic mobility shift assay (EMSA) was used to detect NF-?B DNA binding activity. RESULTS: Homocysteine (0.1-1 mmol/L) decreased the human umbilical vein endothelial cell (HUVEC) viability and increased the levels of ROS. Western blotting and ELISA showed that homocysteine significantly increased the expression of TNF-?, IL-6, MCP-1 and ICAM-1. However, pretreatment with simvastation (1-20 ?mol/L) reversed the decreased cell viability and markedly suppressed an increase in the ROS level and the expression of TNF-?, IL-6, MCP-1 and ICAM-1 induced by homocysteine. Homocysteine induced p38 phosphorylation and such phosphorylation was also inhibited by simvastation and antioxidant NAC. EMSA and Western blotting showed that homocysteine induced NF-?B activation due to the increased phosphorylation of the inhibitory protein (I?B?) as well as the degradation of I?B?, while simvastation pretreatment almost completely blocked the NF-?B activation as well as the phosphorylation and degradation of I?B?. CONCLUSION: Simvastation inhibits homocysteine-induced endothelial dysfunction and inflammatory response through interfering with ROS-p38-NF-?B pathway.

OBJECTIVETo investigate the alteration of inflammasome and receptor during IgG promoter transfected to HepG2 cells.

METHODBy assay of Elisa to evaluate the secretion of IL-1 beta, IL-8, TNF-alpha and MCP-1 after puerarine and LPS administration, and by assay of real time PCR to evaluate the expression of mRNA of IL-1 beta, IL-8,TNF-alpha and MCP-1, as well as the receptors of TLR2, 4 and NOD2, MyD88.

RESULTIgG promoter did not active innate immunity and enhance the expression and secretion of inflammasome in HepG2. Puerarine did not active the inflammasome either. LPS activated the innate immunity and increased the secretion of IL-8, TNF-alpha and MCP-1.

CONCLUSIONIgG-HepG2 cells could be used specifically as the model of allergy type II for ingredients screening. It is suggested that puerarine was suite for the activator for this type of allergy as positive control.

Allergens ; analysis ; immunology ; Drugs, Chinese Herbal ; chemistry ; Gene Expression Regulation ; immunology ; Hep G2 Cells ; Humans ; Immunity, Innate ; immunology ; Immunoglobulin G ; genetics ; Inflammasomes ; immunology ; Promoter Regions, Genetic ; genetics ; Transfection

There have been many reports on berberine (BBR) effect of the inhibition on gut bacteria,but more from the protein level.In view of the preference of BBR for DNA binding,we here investigated the expression of BBR from the transcriptional expression level of the gene.The results showed that BBR had a higher affinity for UP element of Escherichia coli (E.coli) gene,and the transcription initiation region of this element contained TATA base sequence.The expression of genes sulA,recA and 16S which contain the genes of the UP element regulatory elements in the upstream of the promoter could be suppressed by BBR,and the expression of lpxC,secG and mutT which did not contain the genes of the UP element regulatory elements in the upstream of the promoter could not be inhibited by BBR.It is shown that the TATA sequence is the target of BBR.This result provides a new perspective for exploring the effect of BBR's inhibition of microbiota from gene transcription.

Objective To compare the differences and clinical value of prognostic evaluation between American Joint Committee on Cancer (AJCC) TNM staging system 7th edition and 8th edition for gastric cancer (GC).Methods The retrospective case-control study was conducted.The clinicopathological data of 1 383 GC patients who were admitted to the First People's Hospital of Changzhou between January 2008 and August 2012 were collected.Distal gastrectomy,proximal gastrectomy + pyloroplasty or total gastrectomy were performed according to preoperative evaluation and intraoperative exploration.Observation indicators:(1) surgical and postoperative situations;(2) follow-up and survival situations;(3) T staging comparison between AJCC TNM staging system 7th edition and 8th edition;(4) N staging comparison of AJCC TNM staging system 8th edition;(5) prognostic analysis in N staging of AJCC TNM staging system 8th edition;(6) TNM staging comparison between AJCC TNM staging system 7th edition and 8th edition;(7) prognostic analysis in different TNM staging between AJCC TNM staging system 7th edition and 8th edition.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to October 2017.Measurement data with normal distribution were represented as x ± s.Measurement data with skewed distribution were described as M (range).The survival curve and survival rate were respectively drawn and calculated by the Kaplan-Meier method,and the Log-rank test was used for survival analysis.Results (1) Surgical and postoperative situations:1 383 GC patients underwent successful radical gastrectomy,including 923 with distal gastrectomy,165 with proximal gastrectomy and 295 with total gastrectomy.Of 1 383 patients,115 with postoperative complications were improved by symptomatic treatment,including 87 with surgical complications and 28 with non-surgical complications.Postoperative pathological examinations:total number of intraoperative lymph node dissection and number of lymph node metastasis were 25± 12 and 7±4;577 didn't have lymph node metastasis and 806 had regional lymph node metastasis;308 were in early GC and 1 075 in advanced GC.(2) Follow-up and survival situations:1 383 patients were followed up for 1-117 months,with a median time of 34 months.The 1-,3-and 5-year survival rates of 1 383 patients were respectively 90.5％,71.9％ and 61.1％.(3) T staging comparison between AJCC TNM staging system 7th edition and 8th edition:T staging definition between AJCC TNM staging system 7th edition and 8th edition was identical.T staging of 1 383 patients:308,192,65,628 and 190 were respectively detected in T1,T2,T3,T4a and T4b stagings.(4) N staging comparison between AJCC TNM staging system 7th edition and 8th edition:N staging definition between AJCC TNM staging system 7th edition and 8th edition was identical.N staging of 1 383 patients:577,255,207,230 and 114 were respectively detected in N0,N1,N2,N3a and N3b stagings.N3a and N3b were classified as N3 staging of AJCC TNM staging system 7thedition,but they were classified as independent staging of AJCC TNM staging system 8th edition.(5) Prognostic analysis in N staging of AJCC TNM staging system 8th edition:5-year survival rate of patients in N0,N1,N2,N3a and N3b stagings was respectively 85.6％,76.5％,59.4％,45.2％ and 32.5％ based on AJCC TNM staging system 8th edition,with a statistically significant difference in survival (x2 =394.400,P＜0.05).There was a statistically significant difference between N0 and N 1 stagings (x2 =45.630,P＜0.05),between N 1 and N2 stagings (x2 =19.470,P＜0.05),between N2 and N3a stagings (x2 =7.602,P＜0.05) and between N3a and N3b stagings (x2=13.020,P＜0.05).(6) TNM staging comparison between AJCC TNM staging system 7th edition and 8th edition:TNM staging of 366 patients had changes,including 2 in T1N3b staging,2 in T2N3b staging,18 in T3N3b staging,120 in T4aN2 staging,149 in T4aN3a staging,34 in T4bN0 staging and 41 in T4bN2 staging;364 were detected in staging Ⅲ in 7th edition and 8th edition,and sub-staging of staging Ⅲ had a change;2 in T1N3b of ⅡB staging were redistricted into Ⅲ B staging based on AJCC TNM staging system 8th edition.(7) Prognostic analysis in different TNM staging between AJCC TNM staging system 7th edition and 8th edition:according to 7th edition,cases and 5-year survival rate were respectively 247,89.5％ in Ⅰ A staging and 147,83.7％ in Ⅰ B staging and 77,75.9％ in ⅡA staging and 207,70.5％ in ⅡB staging and 136,61.0％ in ⅢA staging and 236,37.5％ in Ⅲ B staging and 333,35.4％ in Ⅲ C staging,with a statistically significant difference in survival among sub-stagings (x2 =228.800,P＜0.05).There was a statistically significant difference in survival among Ⅰ,Ⅱ and Ⅲ stagings (x2=189.000,P＜0.05) and between ⅢA and ⅢB or ⅢC stagings (x2=22.710,18.010,P＜0.05).There was no statistically significant difference in survival between Ⅰ A and Ⅰ B stagings (x2=0.179,P＞0.05),between Ⅱ A and Ⅱ B stagings (x2 =0.265,P＞0.05),and between Ⅲ B and Ⅲ C stagings (x2 =1.550,P＞0.05).According to 8th edition,cases and 5-year survival rate were respectively 247,89.5％ in Ⅰ A staging and 147,83.7％ in Ⅰ B staging and 77,75.9％ in Ⅱ A staging and 205,70.7％ in Ⅱ B staging and 288,53.8％ in ⅢA staging and 258,37.3％ in ⅢB staging and 161,28.5％ in ⅢC staging,with a statistically significant difference in survival among sub-stagings (x2=234.900,P ＜ 0.05).There was no statistically significant difference in survival between Ⅰ A and Ⅰ B stagings (x2 =0.179,P＞0.05) and between Ⅱ A and ⅡB stagings (x2 =0.564,P＞0.05).There was statistically significant differences in survival between Ⅲ A and Ⅲ B or ⅢC stagings (x2 =29.790,43.060,P＜0.05) and between Ⅲ B and Ⅲ C stagings (x2 =7.494,P＜0.05).Further analysis showed that changes of TNM staging system between 7th edition and 8th edition were in T3N3b,T4aN2,T4aN3a,T4bN0 and T4bN2 stagings,5-year survival rate in above stagings was respectively 16.7％,35.8％,30.2％,47.1％ and 26.8％,with statistically significant differences in survival between T3N3b and T4aN2,T4aN3a,T4bN0 and T4bN2 stagings (x2 =19.590,8.039,12.070,3.853,P＜0.05),between T4aN2 and T4aN3a,T4bN2 stagings (x2 =6.529,3.859,P ＜ 0.05),between T4aN3a and T4bN0 stagings (x2 =10.400,P＜0.05) and between T4bN0 and T4bN2 stagings (x2=4.636,P＜0.05).There was no statistically significant difference in survival between T4aN2 and T4bN0 stagings (x2 =3.607,P＞0.05) and between T4aN3a and T4bN2 stagings (x2 =0.029,P＞0.05).Conclusions Compared with AJCC TNM staging system 7th edition,N3a and N3b stagings are classified as independent staging in AJCC TNM staging system 8th edition,and 8th edition is more accurate in prognostic evaluation of GC patients in stage Ⅲ.

OBJECTIVETo study the allergen in key processes during the production of Fufang Kushen injection by IgG promoter-HepG2 cells in vitro.

METHODBy transfecting a IgG promoter-regulating the expression of green fluorescent protein(GFP) plasmid into HepG2 cells, this transferred cells were incubated with common allergens (like puerarin, ovalbumin, LPS or Sal typhoid vi polysaccharide vaccine), excipients using in Fufang Kushen injection (NaOH, acetic acid, Tween-80 and ethanol) and samples from the key production processes of the injection for 30 minutes . Fluorescent photographs were analyzed the fluorescence intensity of the cells by using an image analysis software.

RESULTAll of common allergens significantly increased the IgG expression. Two of the excipicents, acetic acids and Tween-80 were shown to increased the IgG expression, while others had no effect on IgG expression. In the 8 samples from the key processes in the production of Fufang Kushen injection, two of them stimulated IgG expression.

CONCLUSIONIgG promoter-HepG2 cells are highly sensitive and specific to allergens, and thus can be applied to rapid screening of allergens in components and injections in transcriptional level. It is possible to use the IgG-promoter HepG2 cells in a real-time monitoring of allergens in the production processes of Chinese medicine injections.

Allergens ; analysis ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Gene Expression Regulation ; immunology ; Hep G2 Cells ; Humans ; Immunoglobulin G ; genetics ; Injections ; Medicine, Chinese Traditional ; standards ; Promoter Regions, Genetic ; genetics ; Quality Control

Objective To explore the role of clinical pharmacists in the treatment of drug poisoning by analyzing the clinical pharmacist's participation in the treatment of a patient with sodium valproate poisoning. Methods Clinical pharmacists measured the plasma concentration of sodium valproate to inform the doctor to diagnose illnesses. At the initial stage when the concentration is high, to eliminate the free drug by continuous venous-venous hemodialysis-filtration (CVVHDF). Then, the combined drug was cleared by hemoperfusion (HP). Results The blood concentration dropped by half at the first CVVHDF and decreased obviously after two HPs. After stable observation in five days’ course of disease, the blood concentration was maintained at a low level and the patient was cured and discharged. Conclusion The implementation of the blood purification program under the monitoring of the blood drug concentration with the participation of pharmacists is helpful for the rescue of drug overdose and is worthy of promotion.