Objective To investigate the role of Toll-like receptor 4 ( TLR4 ) and NOD-like receptor 3 (NLRP3)inflammasome in the liver injury of acute necrotizing pancreatitis (ANP) rat with obesity. Methods Twenty-four SD rats were randomly divided into normal group, ANP group, obesity group and obesity ANP group. The obesity rat model was established by continuously feeding high fat diet and the ANP model was induced by retrograde infusion of 5％ sodium taurocholate into the biliopancreatic duct. Rats were killed at 12 h after model establishment, and automatic biochemical immune analyzer were used for detecting serum AMY, LIP, ALT, AST, TG and TC. Pathological changes of pancreas and liver tissue samples were observed by miscroscopy and pathological score was recorded. The levels of MPO, CD68 , TLR4, NLRP3 and IL-1βin liver tissue were detected by immunofluorescence, and NF-κB and caspase-3 in liver tissue were detected by immunohistochemistry. Results The serum ALT and AST in obesity ANP group were significantly increased than those in ANP group (233. 00 ± 34. 44 U/L vs 102. 83 ± 8. 90 U/L,388. 00 ± 41. 60 U/L vs 282. 00 ± 21. 06 U/L);and liver pathologic score was also significantly higher than ANP group (6. 66 ± 1. 21 vs 3. 33 ± 1. 03);and CD68 + /TLR4 +, CD68 + /NLRP3 +, TLR4 + /NLRP3 +, MPO, NF-κB, IL-1β and caspase-3 level were all greatly higher in obesity ANP than those in ANP group, respectively (24. 16 ± 1. 47 vs 6. 66 ± 1. 21, 25. 00 ± 2. 60 vs 7. 00 ± 1. 41, 14. 16 ± 1. 47 vs 5. 50 ± 1. 04, 35. 33 ± 6. 88 vs 20. 83 ± 2. 48, 58. 80 ± 6. 75 vs 37. 63 ± 2. 96, 50. 00 ± 2. 36 vs 35. 00 ± 2. 82, 66. 00 ± 4. 04 vs 55. 00 ± 2. 60); and all the differences were statistically significant (all P<0. 05). Conclusions Liver injury was more severe in ANP rats with obesity, which may be related to the fact that obesity may enhance the activation of TLR4/NLRP3 signal pathway and result in the release of more inflammatory factors.

OBJECTIVE： To establish a method for content determination of total polyphenols from Gastrodia elata， and to optimize the purification technology of macroporous resin. METHODS： The content of total polyphenols from G. elata was determined by Folin-ciocaileu colorimetry. Using the absorption and desorption performance as index， 4 kinds of macroporous adsorption resins were selected by static adsorption and desorption tests. The flow rate and mass concentration of the sampling solution， volume fraction of eluent， eluent flow rate and eluent volume were investigated by dynamic adsorption and desorption tests. The purification technology of macroporous resin was optimized. RESULTS： The linear range of gallic acid was 4-32 μg/mL （r＝0.999 9）. RSDs of precision， stability and repeatability tests were all less than 2％. The recovery rate of the sample was 95.51％-102.94％（RSD＝2.54％，n＝6）. D301 macroporous resin had strong static adsorption and desorption ability from G. elata polyphenols. The optimal purification technology included that the sample solution flow rate 2 BV/h； the sample solution mass concentration 4 mg/mL； the elution solvent 70％ ethanol； the elution flow rate was 3 BV/h， and the eluent volume 5 BV. The content of total polyphenols from G. elata optimized by the optimal purification technology was 0.381 mg/g. CONCLUSIONS： Established method is sensitive and stable. The optimized purification technology is stable and feasible.

Objective: To study the mRNA expressions of various CD97 isoforms in colorectal carcinoma tissues and their clinical significances. Methods: A total of 50 colon cancer patients in the First Affiliated Hospital of Wenzhou Medical University from December 2013 to May 2014 and human colon cancer cell lines SW480 and SW620 were enrolled. The real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expressions of CD97 human epidermal growth factor (EGF) (1, 2, 5), CD97EGF (1, 2, 3, 5) and CD97EGF (1, 2, 3, 4, 5) in colon cancer tissues, adjacent tissues, normal colon tissues, SW480 cells and SW620 cells. The relationship between the mRNA expression of CD97EGF (1, 2, 5) and the clinicopathological factors was analyzed. Results: Compared with those low expressions in adjacent tissues and normal tissues, the mRNA expressions of CD97 isoforms CD97EGF (1, 2, 5), CD97EGF (1, 2, 3, 5) and CD97EGF (1, 2, 3, 4, 5) in cancer tissues were highest, and the differences were statistically significant (0.71±0.20 vs. 0.40±0.09 vs. 0.35±0.07, F = 107.642, P < 0.01; 0.45±0.11 vs. 0.26±0.05 vs. 0.27±0.06, F = 94.231, P < 0.01; 0.41±0.10 vs. 0.21±0.05 vs. 0.19±0.03, F = 165.672, P < 0.01). In addition, the mRNA expression of CD97EGF (1, 2, 5) in colon cancer patients was associated with tumor infiltration depth (T₁-T₂ and T₃-T₄), clinical stages (Ⅰ-Ⅱ and Ⅲ-Ⅳ), and the differences were statistically significant (t = -2.582, P = 0.013; t = -5.062, P < 0.01). The mRNA expression of CD97EGF (1, 2, 5) in SW620 cells was higher than that in SW480 cells. Conclusions CD97 isoforms are highly expressed in colon cancer tissues, and CD97EGF (1, 2, 5) may play an important role in the development and invasion of colon cancer. The CD97 isoforms may be new markers in the treatment of colon cancer.

Objective To compare the diagnostic accuracies of deep convolutional neural network (CNN) and dermatologists for pigmented nevus and seborrheic keratosis.Methods CNN network ResNet-50 was trained with 5 094 dermoscopic images of pigmented nevus and seborrheic keratosis using transfer learning,so as to establish a CNN two-classification model.Then,this model was applied to the automatic classification of 30 dermoscopic images of pigmented nevus and 30 dermoscopic images of seborrheic keratosis.Meanwhile,in combination with clinical photos of skin lesions,95 experienced dermatologists who had received dermoscopy training gave their diagnosis for the above 60 dermoscopic images.The diagnostic accuracies were compared between the two methods,and misclassified images were further analyzed.Results The CNN automatic classification model had the diagnostic accuracies of 100％ (30/30)and 76.67％ (23/30) for pigmented nevus and seborrheic keratosis respectively,and the total accuracy was 88.33％ (53/60).The average diagnostic accuracies of 95 dermatologists were 82.98％ (25.8/30) and 85.96％ (24.9/30) for pigmented nevus and seborrheic keratosis respectively,and the total accuracy was 84.47％ (50.7/60).There were no significant differences in the diagnostic accuracies for pigmented nevus or seborrheic keratosis between the CNN automatic classification model and 95 dermatologists (x2 =0.38,P ＞ 0.05).The dermoscopic images misclassified by CNN were divided into 3 categories:special-type lesions with high pigment content and marked keratosis,typical skin lesions with interference factors,and typical skin lesions without definite reasons for misclassification.Conclusions The performance of CNN automatic classification model is similar to that of experienced dermatologists in the two classification of pigmented nevus and seborrheic keratosis.The reasons for misclassification by CNN still need to be explored by dermatologists and professionals in artificial intelligence.

Objective To report a pedigree with X?linked dominant protoporphyria(XLDPP), and to detect 5?aminolevulinic acid synthetase 2(ALAS2)gene mutations in this pedigree. Methods A clinical investigation was performed in a pedigree with XLDPP, and relevant data were collected from family members. A next?generation sequencing method was applied to screen possible mutation sites, and Sanger sequencing was performed to determine pathogenic gene mutations. Dermoscopy was conducted to observe skin lesions in the patients with XLDPP, and the Fotofinder system and very high frequency (VHF) ultrasound system were utilized to assess the severity of photodamage. Liver and gallbladder ultrasonography as well as blood examination were performed for all the family members. Results A deletion mutation, c.1706?1709ΔAGTG, was detected in the ALAS2 gene on the X chromosomes of all the patients in this family, which led to replacement or loss of 19-20 C?terminal residues through transcriptional frameshifting, and eventually caused an increase in ALAS2 activity. In the patients with XLDPP, skin photodamage was relatively severe;protoporphyrin?induced hepatobiliary damage was observed and aggravated with age;anemia and iron deficiency occurred sometimes. Conclusion The deletion mutation c.1706?1709ΔAGTG of the ALAS2 gene may be the underlying cause of XLDPP in this pedigree.

AIM:To investigate the role of heat shock protein 70(HSP70)in hepatitis B virus (HBV) replica-tion.METHODS:The effect of HBV replication on the expression of HSP 70 was analyzed by RT-qPCR.The overexpres-sion efficiency of HSP70 was confirmed by Western blot .The effect of HSP70 overexpression on HBV DNA replicative in-termediates was analyzed by RT-qPCR and Southern blot .The effects of HSP70 overexpression on the expression level of HBV 3.5 kb mRNA and HBV core protein were measured by RT-qPCR and Western blot, respectively.The Effect of HSP70 overexpression on HBV promoter activity was detected by dual luciferase reporter system .RESULTS: The mRNA levels of HSP70 were inhibited by HBV replication .Overexpression of HSP70 repressed the expression of HBV DNA repli-cative intermediates, 3.5 kb mRNA and core protein, as well as HBV core promoter activity .CONCLUSION:HBV rep-lication inhibits the expression of HSP70.Overexpression of HSP70 represses HBV replication.These data suggest that HSP70 repressed HBV replication by inhibiting HBV core promoter activity .

OBJECTIVETo analyze the clinical efficacy and toxicity of vitamin support in lung adenocarcinoma patients treated with pemetrexed second-line chemotherapy.

METHODSTwo hundred and eighty-three patients with stage 3/4 lung adenocarcinoma treated at our hospital from August 2010 to August 2013 were included in this study. The lung adenocarcinomas in all the 283 patients were confirmed by pathology or cytology, all were EGFR-negative, and all patients received pemetrexed second line chemotherapy. The 283 patients were randomly divided into two groups: the improved treatment group (142 cases) and the conventional treatment group (141 cases). The patients of conventional treatment group received 400 µg folic acid per os daily for 7 days before the first dose of pemetrexed, and continued until 21 days after the last dose of pemetrexed. Besides, they received 1000 µg vitamin B12 injection at 7 days before the first dose of pemetrexed, and once per cycle of pemetrexed for 3 cycles after the last dose of pemetrexed. The patients of the improved treatment group took 400 µg folic acid daily per os from the day before the first dose to 21 days after the last dose of pemetrexed. They also received 500 µg vitamin B12 by injection one day before the first dose, and one day before each therapy cycle of pemetrexed therapy.

RESULTSThe mean number of cycles of pemetrexed chemotherapy was 4 in both groups. In the 142 patients of improved treatment group, complete response (CR) was observed in two cases, partial remission (PR) in 28, stable disease (SD) in 21, and progressive disease (PD) in 91 cases, with a total effective rate of 21.1%. While in the conventional treatment group, CR was observed in one case, PR in 27 cases, SD in 23 cases, and PD in 90 cases, with a total effective rate of 19.9%. The median progression-free survival (PFS) was 3.8 months in the improved treatment group and 4.2 months in the conventional treatment group (P=0.143). The toxicity of chemotherapy was mild in both groups, with no significant difference between the two groups (P>0.05). The most common side effects of hematological system were leukopenia and neutropenia, and the most common side effects of non-blood system were nausea and vomiting. The most common grade 3-4 toxic reaction in both groups was leukopenia and neutropenia, with no significant difference between the two groups (P>0.05). Multivariate analysis showed that the age of patients was an independent factor of grade 3-4 chemotherapy toxic reaction (P<0.05), while gender, the baseline level of PS score or blood system had no significant effect on the grade 3-4 chemotherapy toxic reaction (P>0.05).

CONCLUSIONSCompared with the conventional treatment scheme, the improved treatment scheme has similar therapeutic effects and could be used more conveniently, while the toxic effects of chemotherapy are not increased at the same time. Our results indicate that pemetrexed-based chemotherapy does not need to delay the chemotherapy because of vitamin support treatment.

Adenocarcinoma ; drug therapy ; Antineoplastic Agents ; therapeutic use ; Disease-Free Survival ; Folic Acid ; therapeutic use ; Humans ; Lung Neoplasms ; drug therapy ; Pemetrexed ; therapeutic use ; Treatment Outcome ; Vitamin B 12 ; therapeutic use ; Vitamin B Complex ; therapeutic use

OBJECTIVETo research the effect of Jianpi Jiedu decoction (JPJDT) to PTEN/ERK1 of athymic mice with hepatocellular carcinoma.

METHODN2 male BALB/c athymic mice models were built by Bel-7402 with an indirect method. After 24 h of postoperation, the 90 athymic mice were distributed randomly into JPJDT groups: A, B, C, D, E, F, G, NS, FT each group had 10 athymic mice. Another 10 male BALB/c athymic mice without HCC was treated by NS as normal control (DZ). Group A to G were treated by intragastric administration with JPJDT that had been deliquated into 7 kinds of density for 8 wk. Group NS were were treated by intragastric administration with Sodium Chloride for 8 wk. Group FT were were treated by intragastric administration with FT207 (tegafur) for 8 wk . At last, athymic mice were sacrificed. PTEN/ERK1 was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision two-step histostaining reagent).

RESULTThe expression intensity of PTEN: The result showed that the intensity of PTEN in the normal hepatic tissue was the highest, and then latero-cancer tissue, the lowest was cancer tissue. In the normal hepatic tissue, the intensity of PTEN in Group B, D, E was higher than the Group NS, Group FT, Group DZ (P < 0.05). In the latero-cancer tissue, the intensity of PTEN in Group D was higher than the Group NS (P < 0.05). In the cancer tissue, the intensity of PTEN in Group JPJDT was higher than the Group NS and Group FT (P < 0.05). The expression intensity of ERK1: The result showed that the intensity of PTEN in the cancer tissue was the highest, and then latero-cancer tissue, the lowest was normal hepatic tissue. In the latero-cancer tissue, the intensity of ERK1 in Group FT was higher than the Group NS and Group JPJDT (P < 0.05). In the cancer tissue, the intensity of PTEN in Group NS and Group FT was higher than the Group C, D, E, G, F (P < 0.05). The correlation between PTEN and ERK1: The result showed that there was inverse correlation between the expression intensity of PTEN and ERK1 in the cancer tissue (P < 0.01).

CONCLUSIONOne of mechanism of antitumous effect of JPJDT maybe up-regulate anti-oncogene PTEN, restrain the signal way of ERK1, suppress the proliferation of hepatoma carcinoma cell. The carcinogenesis of primary hepatic carcinoma may exist the deletion of PTEN. Owing to low expression or deletion of PTEN in the cancer tissue, ERK1 signal transduction pathway cannot be actively suppressed which was activated by carcinogenic factor. So hepatoma carcinoma cell multiplicated.

Animals ; Carcinoma, Hepatocellular ; genetics ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Male ; Mice ; Mice, Nude ; Mitogen-Activated Protein Kinase 3 ; metabolism ; PTEN Phosphohydrolase ; metabolism

Objective To investigate the perioperative management for patients of thyroid crisis complicating acute abdomen.Methods The clinical data of 11 acute abdomen patients combined with hyperthyroidism were analyzed retrospectively.Results In this study,the surgical procedures were appendectomy in 5 patients,perforation repair for duodenal ulcer in 2 patients,distal gastrectomy in 2 patients,choledochotomy in one patient and splenectomy in one patient.Thyroid crisis occurred postoperatively in 7 patients,intraoperatively in 2 patients and preoperatively in 2 patients.Emergency treatment succeeded in 8 patients,and death occurred in 3 patients,with a death rate of 27.3%,including the preoperative thyroid crisis patient complicating duodenal ulcer perforation,who died of heart sufficiency on the first day postoperation.The second patient suffering from intraoperative thyroid crisis during exploration for perforation of the small bowel died of muhiple organ dysfunction syndrome caused by intestinal fistula,intra-abdominal abscess and lung infection,in spite of successful rescue treatment of thyroid crisis.The third patient was of acute appendicitis with perforation who died of coma and multiple organ dysfunction syndrome on the postoperative 5th day because the concomitant thyroid crisis was overlooked.Conclusions The early recognizing thyroid crisis in perioperative surgical abdomen and adequate treatment is the key to rescue succesS,and the diagnosis mainly depends on comprehensive judgment of clinical presentation and laboratory examination.

Objective To evaluate the effect of bronchoscopic argon plasma coagulation thera-py on bronchial carcinoma. Methods Thirty-one bronchial carcinoma patients were diagnosed by bronchoscope and pathological tests, with or without atelectasis or obstructive pneumonia on chest X-ray or chest CT. Argon plasma coagulation therapy was performed through bronchoscope. The location of the airway lesions, the degree of obstruction, dyspnea index, and complications were evaluated. Results The patients with bronchial carcinoma were treated 1～4 times by bronchoscopic argon plas-ma coagulation therapy. Full effectiveness was achieved in 15 patients (48.4 %), partial in 12 (38.7%), and mild in the other 4 ( 12.9 % ). The overall effective rate was 100 %. Conclu-sion Bronchoscopic argon plasma coagulation therapy for bronchial carcinoma can remarkably reduce the tumor size, relieve clinical symptoms, and alleviate the obstruction caused by bronchial neoplasm. Brouchoscopic argon plasma coagulation therapy is an effective and safe method for patients with bron-chial carcinoma.