For wild natural medicine, unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials, which affects the efficacy and safety of clinical medication. DNA barcoding as an effective species identification tool is limited by its low sample throughput nature. In this study, combining DNA mini-barcode, DNA metabarcoding and species delimitation method, a novel biological sources consistency evaluation strategy was proposed, and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as "Guang Dilong" and 25 batches of proprietary Chinese medicines. Besides Amynthas aspergillum as the authentic source, 8 other Molecular Operational Taxonomic Units (MOTUs) were elucidated. Significantly, even the subgroups within A. aspergillum revealed here differ significantly on chemical compositions and biological activity. Fortunately, this biodiversity could be controlled when the collection was limited to designated areas, as proved by 2796 "decoction pieces" samples. This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control, and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.

Objective:To explore the long-term effect of endoscopic radiofrequency ablation for the treatment of unresectable ampullary carcinoma.Methods:Clinical data of patients with ampullary carcinoma who received endoscopic radiofrequency ablation in the Third Affiliated Hospital of Naval Medical University from January 2012 to May 2019 were retrospectively collected, including basic information, frequency of radiofrequency ablation, the type of biliary stent, postoperative complications, and follow-up. Kaplan-Meier method was used to analyze the survival of patients after endoscopic radiofrequency ablation. Relationship between frequency of radiofrequency ablation, type of biliary stent and overall survival time was analyzed.Results:A total of 50 patients were enrolled, including 31 males and 19 females, aged 73.0±9.7 years. Twenty-five patients (50.0%) underwent 1 radiofrequency ablation treatment, while 25 patients (50.0%) underwent radiofrequency ablation treatments more than twice. Postoperative complications occurred in 6 patients (12.0%), all of which were mild symptoms. The average follow-up was 22.3 months, with a total of 39 (78.0%) deaths, 5 (10.0%) lost to follow-up, and 6 (12.0%) surviving. The median overall survival time was 16.9 (95% CI: 9.1-24.8) months, with cumulative survival rates of 62.0%, 38.5%, 27.0%, and 12.6% at 1, 2, 3, and 5 years, respectively. The median overall survival time of those treated with radiofrequency ablation ≥2 times showed a trend of prolongation compared to patients treated once, but the difference was not statistically significant [26.7 (95% CI: 9.7-43.7) months VS 12.6 (95% CI: 4.9-20.3) months, χ2=3.049, P=0.081]. Plastic stents were used in 32 patients (64.0%) and metal stents in 18 patients (36.0%). There was no significant difference in median overall survival time between patients using metal and plastic stents [17.1 (95% CI: 6.1-28.0) months VS 15.9 (95% CI: 6.9-24.9) months， χ2=0.029， P=0.865]. Conclusion:Endoscopic radiofrequency ablation is a safe treatment for unresectable ampullary carcinoma, and multiple consecutive treatments may increase the survival benefit.

Objective:To investigate the efficacy of endoscopic stent placement for patients with Bismuth type Ⅳ hilar cholangiocarcinoma.Methods:Data of 229 patients with unresectable Bismuth type Ⅳ hilar cholangiocarcinoma who successfully underwent endoscopic stent placement at the Department of Endoscopy, the Third Affiliated Hospital of Naval Medical University from January 2002 to January 2019 were retrospectively analyzed. Outcomes included clinical success rate, complication incidence, stent patency period and overall survival time. The patency of stents and overall survival time of patients were estimated by using the Kaplan-Meier method. The independent predictors for stent patency and overall survival of patients were analyzed by a multivariate Cox proportional regression model.Results:The overall clinical success rate was 78.2% (179/229). The incidence of early cholangitis after endoscopic retrograde cholangiopancreatography was 20.5% (47/229). The median stent patency and overall survival time were 5.7 (95% CI: 4.8-6.7) months and 5.1 (95% CI: 4.2-6.0) months, respectively. Further multivariate Cox regression analysis showed that metal stent ( P<0.001, HR=0.452, 95% CI: 0.307-0.666) and bilateral stents with bilateral angiography ( P=0.036, HR=0.644, 95% CI: 0.427-0.971) were independent predictors of stent patency; total bilirubin>200 μmol/L ( P=0.001, HR=1.627, 95% CI: 1.208-2.192), metal stent ( P=0.004, HR=0.636, 95% CI: 0.467-0.866) and antitumor therapy ( P<0.001, HR=0.439, 95% CI:0.308-0.626) were independent predictors of overall survival. Conclusion:There is high incidence of cholangitis in patients with unresectable Bismuth type Ⅳ hilar cholangiocarcinoma treated with endoscopic stenting. Longer stent patency can be achieved with metal stent placement and bilateral drainage. In addition, metal stent for drainage and antitumor therapy can also help increase the survival benefit.

Objective:To analyze the expression of E-cadherin in the epithelial-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1) in neuroblastoma.Methods:TGF-β1(1 μg/L, 5 μg/L, 10 μg/L), was applied to SK-N-SH cells in vitro compared with the blank control group.EMT-related genes mRNA and protein expression were detected by carrying out real-time PCR assays and Western blot.A scratch test and migration assay were performed to verify the alteration of SK-N-SH cell migration capacity.Data collected from 18 cases of neuroblastoma patients were selected from the Department of Hematology Oncology, Shanghai Children′s Hospital from January 2008 to December 2012.The expression of E-cadherin in the tumor tissue of the neuroblastoma patients after operation was detected by immunohistochemistry.The clinical features and survival prognosis of these patients were analyzed. Results:Compared with the control group, after SK-N-SH cells were treated with TGF-β1(1 μg/L, 5 μg/L, 10 μg/L), real-time PCR assays and Western blot revealed that the mRNA(0.603±0.081, 0.606±0.008, 0.716±0.166 vs.1.000) and protein expression levels(0.855±0.026, 0.600±0.017, 0.495±0.011 vs.1.000) of E-cadherin were significantly decreased ( F=8.144, P=0.040; F=74.810, P<0.001), while the mRNA(2.132±0.167, 3.494±0.017, 4.184±0.021 vs.1.000) and protein expression levels (1.175±0.053, 1.189±0.058, 1.225±0.106 vs.1.000)of α - smooth muscle actin were significantly increased ( F=547.300, P<0.001; F=68.810, P=0.007), suggesting that EMT changes occur in cells.Scratch test and Transwell migration assay revealed that the number of migrating cells increased obvious with the treatment of TGF-β1 (5 μg/L) ( t=16.070, P=0.040). The 10-year overall survival(OS) rates of neuroblastoma patients with E-cadherin strong positive expression, positive expression, weak positive expression and negative expression in the pathology were (77.78±13.86)%, (75.00±21.66)%, (25.00±21.65)% and 0, respectively ( F=8.160, P=0.040). Conclusions:TGF-β1 can induce the EMT in SK-N-SH cells and increase cell migration.The decrease expression of E-cadherin in neuroblastoma patients is closely associated with clinical progress and recurrence or metastasis of the disease.

Objective To evaluate the technical feasibility and safety of endoscopic retrograde catheterization of gallbladder (ERCG) and endoscopic transpapillary gallbladder stenting (ETGS) for gallbladder diseases.Methods Patients who underwent ERCG and ETGS in Eastern Hepatobiliary Hospital from January 2010 to June 2016 were enrolled to this retrospective study.The superselection time of cystic duct,the catheterization time of gallbladder,postoperative symptoms and complications were analyzed.Results A total of 10 patients were enrolled to this study,including 2 cases of acute calculous cholecystitis,4 cases of percutaneous transhepatic gallbladder drainage (PTGBD) and 4 cases of cholecystocholedocholithiasis.The success rates of ERCG and ETGS were 100％.Symptoms were relieved in all patients and PTGBD catheter was removed after ETGS.The mean times of ERCG and ETGS were 10.2 ± 6.9 min and 17.0 ± 8.0 min respectively.The mean times of ERCG were 18.5±4.9 min,13.0±3.6 min and 3.3± 1.3 min,respectively (F=18.86,P =0.002).The mean times of ETGS were 25.5±4.9 min,21.0± 4.7 min and 8.8 ± 1.0 min,respectively (F =18.04,P =0.002).Complications included 1 case of cholangitis and 1 case of hyperamylasemia.Cholangitis was relieved after anti-inflammatory treatment.No acute pancreatitis,bleeding,perforation or procedure-related death occurred.Conclusion ERCG and ETGS are safe and feasible,which can play important roles in the treatment of specific gallbladder diseases or gallbladder with bile duct diseases.

This study aims to evaluate the ability of C-arm cone-beam CT to detect intracranial hematomas in canine models. Twenty one healthy canines were divided into seven groups and each group had three animals. Autologous blood and contrast agent (3 mL) were slowly injected into the left/right frontal lobes of each animal. Canines in the first group, the control group, were only injected with autologous blood without contrast agent. Each animal in all the 7 groups was scanned with C-arm cone-beam CT and multislice computed tomography (MSCT) after 5 minutes. The attenuation values and their standard deviations of the hematoma and uniformed brain tissues were measured to calculate the image noise, signal to noise ratio (SNR) and contrast to noise ratio (CNR). A scale with scores 1-3 was used to rate the quality of the reconstructed image of different hematoma as a subjective evaluation, and all the experimental data were processed with statistical treatment. The results revealed that when the density of hematoma was less than 65 HU, hematomata were not very clear on C-arm CT images, and when the density of hematoma was more than 65 HU, hematomata showed clearly on both C-arm CT and MSCT images and the scores of them were close. The coherence between the two physicians was very reliable. The same results were obtained with C-arm cone-beam CT and MSCT grades in measuring SD value, SNR, and CNR. The reasonable choice of density detection range of intracranial hematoma with C-arm cone-beam CT could be effectively applied to monitoring the intracranial hemorrhage during interventional diagnosis and treatment.
Animals ; Cone-Beam Computed Tomography ; Disease Models, Animal ; Dogs ; Hematoma ; diagnosis ; Image Processing, Computer-Assisted ; Intracranial Hemorrhages ; diagnosis ; Multidetector Computed Tomography ; Signal-To-Noise Ratio

Objective To investigate the image quality difference of the virtual monochromatic spectral(VMS) images synthesized from fast kilovoltage switching and dual-source dual-energy CT for a given radiation dose. Methods A plurality of disposable syringes containing 15 mg/ml iodine contrast agent and saline were placed on the surface of the male anthropomorphic phantom consisting ofhead neck and torso, GE HD750 gemstone spectral imaging and Siemens Somatom Definition Flash dual energy CT protocols were performed on the phantom for a same dose(volume CT dose index 6.52 mGy), and VMS images (40, 60, 70, 80, 100, 120, 140 keV) were reconstructed and obtained(VMSkV switching and VMSdual-source image), respectively. The objective image noise, iodine signal noise ratio(SNR), iodine contrast noise ratio (CNR) and CT values were measured. The results were analyzed using the paired t test and ANOVA. Results All VMS iodine CT values were gradually decreased with the increasing of keV , and iodine CT values on VMSdual-source images were greater than on VMSkV switching images(P<0.05), VMSkV switching iodine CT values in descending order from the head , chest, abdomen at less than 100 keV,while most VMSdual-source iodine CT values were highest on chest(P<0.05). VMSdual-source and VMSkV switching image noise were highest at 40 keV and successively raised from the head, chest , abdomen. VMSkV switching image noise gradually decreased with the increase of keV in the range of 40 to 70 keV , gradually increased up to the trough after 70 keV from 80 keV gradually decreased, while VMSdual-source image noise was lowest at 70 keV or 80 keV , and then with the keV increased gradually increased. Image noise on VMSkV switching images in the range of 40 to 100 keV were higher than that on VMSdual-source images and lower in the range of 100 to 140 keV at most part(P<0.05). VMSdual-source and VMSkV switching iodine SNR were highest at 40 keV or 60 keV,SNR on VMSdual-source images are greater than on VMSkV switching images except 40 keV images on head and abdomen(P<0.05) . VMSdual-source and VMSkV switching iodine CNR were highest at 60 keV,CNR on VMSdual-source images are greater than on VMSkV switching images except 40 keV images on head 100 keV and 120 keV on abdomen(P<0.05).SNR and CNR decreased from the head, chest, abdomen (P<0.05). Conclusion VMS images synthesized from fast kilovoltage switching and dual-source dual-energy CT have different image quality performance at most keV and body parts, but VMS imaging at approximately 70 keV yielded lower image noise, and at approximately 60 keV yielded highest SNR and CNR.

Objective To evaluate the safety and feasibility of shorterning fasting time after gastros-copy in patients with hepato-biliary disease.Methods A total of 300 inpatient cases of hepato-biliary disea-ses who underwent gastroscopy were divided into the treatment group (n =150)and the control group (n =150)using randomized block design.Both groups included three blocks:patients with esophageal and /or gastric varices (n =50),patients undergoing biopsy (n =50)and non-esophageal gastric varices patients without biopsy (n =50)according to the results of gastroscopy.Patients of both groups were asked to eat 2 hours later and drink 30 minutes later after gastroscopy respectively.The symptoms of thirst,hunger,hypo-glycemia,choking cough after eating,aspiration,nausea and vomiting,hematemesis and melena were recor-ded and compared between the two groups.Results There were no significant differences in the incidences between the two groups in thirst and hunger.However the incidence of hypoglycemia was significantly lower in the treatment group than in the control group [(2.0%(3 /150)VS 9.3%(14 /150),P =0.006].And there were also no significant differences in the incidences of choking cough,aspiration,nausea and vomiting 5-8 hours after the procedure.Neither hematemesis nor melena was found in either groups 24 hours after the procedure.Conclusion Shortening the fasting time after gastroscopy in patients with hepato-biliary diseases is safe and feasible.

Objective To evaluate the effectiveness of mouth care combined with intestinal and endoscopic working channel flushing for ERCP related cholangitis and to investigate the mechanisms.Methods A total of 284 patients who underwent ERCP were divided into the amikacin group (n =96),the saline group (n =95) and the control group (n =93).Specimens were collected before cannulation (Sample 1),the first access to the bile duct (Sample 2) and at the end of ERCP (Sample 3).All the samples underwent bacterial culture and colony counting.Incidence of ERCP related cholangitis and bacterial results were recorded.Results The incidences of ERCP related cholangitis were 7.3％ (7/96),8.4％ (8/95)and 18.3％ (17/93)in amikacin group,saline group and control group respectively,with significant differences among three groups (x2 =6.861,P =0.032).The positive rate of bacterial culture of Sample 1 in the amikacin group,saline group and control group were 6.3％ (6/96),25.3％ (24/95) and 28.0％ (26/93) (x2 =16.833,P =0.000),respectively.The positive rate of bacterial culture of Sample 2 in amikacin group,saline group and control group were 6.3％ (6/96),16.8％ (16/95) and 24.7％ (23/93)(x2 =12.206,P =0.002),respectively.The positive rate of bacterial culture of Sample 3 in amikacin group,saline group and control group were 7.3％ (7/96),17.9％ (17/95) and 33.3％ (31/93) (x2 =20.713,P =0.000),respectively.Conclusion The procedure of ERCP can bring the intestinal bacteria into the bile duct,which could be reduced by mouth care combined with intestinal and endoscopic working channel flushing.And the incidence of ERCP related cholangitis can be effectively reduced.

Objective To clarify the association between rs1050450 polymorphism in Glutathione peroxidase-1 (GPx-1) and the risk of cardi-ovascular diseases (CVD) by performing a meta-analysis of published studies. There is growing evidence from different study types for an association of the GPx-1 polymorphism and cardiovascular outcomes, but observational studies have so far shown inconsistent results. Me-thods Relevant publications were searched through PubMed, Embase database databases and the Cochrane Library. We used odds ratios (ORs) with 95%confidence intervals (CIs) to assess the strength of association under the best genetic model. Both Q statistic and the I2 were used to check heterogeneity. Meta-regression analysis was performed to explore heterogeneity source. Sensitivity analysis, cumulative me-ta-analysis analysis and publication bias were used to test the reliability of the results. Results Data were available from two cohort studies and 8 case-control studies involving 1,430 cases and 3,767 controls. The pooled ORs for overall CVD risk was 1.36 with 95%CI:1.08-1.70 under a co-dominant model, and that for East Asian subgroup was 1.84 (95%CI:1.39-2.43). Substantial heterogeneity for ORs were de-tected among all the included studies, mainly caused by ethnic differences between East Asian and non-East Asian populations. Although Egger’s regression test suggested no statistical significant publication bias, Begg’s funnel plot exhibited obvious asymmetry. The statistical significance disappeared after adjusting for potential publication bias in the overall studies. However, no substantial publication bias was found in the East Asian subgroup. Conclusions GPx-1 gene Pro198Leu and Pro197Leu polymorphisms considerably increased the risk of CVD in the East Asian population. Large-scale investigations are needed to confirm the results in different ethnicities.