[Rapid detection of zearalenone in Coicis Semen based on ELISA].

Zearalenone(ZEN) is a toxic metabolite produced by Fusarium culmorum, F. graminearum, F. tricinctum, and other fungi, with estrogenic characteristics. Exposure to or ingestion of ZEN during pregnancy can cause reproductive dysfunction, miscarriage, stillbirth, and malformation, and seriously endanger human life and health. The detection methods for ZEN in the Chinese Pharmacopoeia(2020 edition) are liquid chromatography(LC) and liquid chromatography-mass spectrometry(LC-MS), and it is stipulated that ZEN should not exceed 500 µg in 1 000 g of Coicis Semen. Although these detection methods by instruments can achieve the qualitative and quantitative analysis of ZEN in Coicis Semen, their high detection cost and long periods hinder the rapid screening of a large number of samples in the field. In this study, the synthesized ZEN hapten was conjugated with bovine serum albumin(BSA) and ovalbumin(OVA) to obtain the complete ZEN antigen. By virtue of antibody preparation techniques, ZEN monoclonal antibody 4F6 was prepared, which showed 177.5%, 137.1%, and 109.7% cross-reactivity with ZEN structural analogs zearalanol, zearalenone, and α-zearalenol, respectively, and no cross-reactivity with other fungal toxins such as aflatoxin. Direct competitive enzyme-linked immunosorbent assay(dcELISA) based on ZEN monoclonal antibody 4F6 was developed for the determination of ZEN in Coicis Semen with an IC_(50) of 1.3 µg·L~(-1) and a detection range of 0.22-21.92 µg·L~(-1). The recoveries were 83.91%-105.3% and the RSD was 4.4%-8.0%. The established dcELISA method was used to determine the ZEN residuals in nine batches of Coicis Semen samples, and the results were validated by LC-MS. The correlation between the two detection methods was found to be 0.993 9, indicating that the established dcELISA could be used for the rapid qualitative and quantitative detection of ZEN residuals in Coicis Semen.

[Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for determination of vomitoxin (DON) in Coicis Semen].

Coicis Semen is a common bulk medicinal material used for both medicine and food, which has the effect of promoting diuresis, draining dampness, invigorating the spleen and checking diarrhea. It is derived from Coix lacryma-jobi var. ma-yuen of the family Poaceae, and is easily contaminated by fungi such as Fusarium graminearum and F. flavum due to climate reasons to produce vomitoxin. The guiding principles for determination of vomitoxin in Chinese medicinal materials in Chinese Pharmacopoeia are mainly HPLC and LC-MS, which have long detection period and are time-consuming and laborious, and thus cannot meet the requirements of on-site quality inspection of drugs. The complete antigen of vomitoxin-protein was obtained by chemical derivatization of vomitoxin. The monoclonal antibody against vomitoxin was prepared by classic monoclonal antibody preparation technology, and enzyme-linked immunosorbent assay(ELISA) method for detection of vomitoxin in Coicis Semen was established through methodological investigation. The IC_(50) based on the ELISA for vomitoxin in Coicis Semen was 3.88 µg·L~(-1), and the average recoveries and the RSD were 77.32%-93.73% and 4.6%-9.7%, respectively. With the established ELISA method, the vomitoxin residue in 14 batches of Coicis Semen samples were determined and validated by LC-MS, and the correlation between the two assays was found to be 0.997 8, indicating that the established ELISA method could be used for quantitative determination of vomitoxin residue in Coicis Semen and could achieve the rapid quantitative determination of the vomitoxin residue.

[Effect of Gansui Banxia Tang plus-minus Gansui and Gancao anti-drug combination that preferred dose close to clinical application on diuretic effect in malignant ascites rats].

OBJECTIVE: To observe the effect of Gansui Banxia Tang plus-minus Gansui and Gancao anti-drug combination on hepatic and renal functions in malignant ascites rats to explore whether the efficacy or toxicity associated with the anti-drug combination. METHOD: The male wistar rats were randomly divided into a blank group, model group, furosemide group, Gansui Banxia Tang group, Gansui Banxia Tang removed Zhigancao group, Gansui Banxia Tang removed Cugansui group, Gansui Banxia Tang removed Zhigancao and Cugansui group. In addition to normal feeding, every morning except for the blank group and model group, the rest of the group was given drugs, the control group and the model group was given distilled water, the volume is 10 mL x kg(-1). Administered five days, all rats were fasted but except water for 24 hours to collect urine. Administered nine days all rats were fasted but except water for 12 hours, we need to weigh weight of rats. When we remove the ascites, we also need to weigh weight of rats. We use the weight before removing ascites minus weight after removing ascites to indirectly measure the amount of ascites. When we remove the ascites, we need to abdominal aortic blood, centrifuge testing renin, angiotensin II, aldosterone, antidiuretic hormone and other indicators. RESULT: The effect of Gansui Banixa Tang on increasing the net weight, lowering abdominal circumference and body weight ratio, lowering renin, angiotensin, aldosterone, antidiuretic hormone is better than the other treatment group. CONCLUSION: In diuresis party, the group of Gansui Banxia Tang is better than the group of Gansui Banxia Tang remove Zhigancao or Cugansui or Zhigancao and Cugansui, renin-angiotensin-aldosterone system may play a diuretic effect of its one way.

Fuji Intelligent Chromo Endoscopy and staining technique for the diagnosis of colon tumor.

BACKGROUND: Colon cancer is a common malignant tumor in the clinic with an incidence rate that is increasing in recent years. The key point for improving the survival rate is the diagnosis and treatment at an early stage. The purpose of this study was to compare the difference of the Fuji Intelligent Chromo Endoscopy (FICE) and staining technique for the diagnosis of colon tumors and non-tumor lesions. METHODS: From March to November 2007, 654 patients were examined with ordinary colonoscopy. Among them 223 patients with colon neoplasm or polypoid lesion were included. The patients were examined with a magnifying ordinary colonoscopy, a magnifying FICE technique and magnifying staining technique. The pit pattern and blood capillary form of the lesion were examined, an endoscopic diagnosis was made and it was compared with the pathologic diagnosis. RESULTS: Four hundred and fifty-one neoplasms were detected in the 223 patients, among those 91.1% (411/451) were detected with the magnifying ordinary endoscopy while 99.1% (447/451) were detected with the FICE technique; there was a significant difference between the two methods. FICE could clearly show the structure and form of mucosal blood capillaries (P < 0.01) but there was no significant difference between the two methods for showing the pit pattern. The coincident rate of FICE for the diagnosis of tumor and non-tumor lesions was 91.6% (413/451), that of the magnifying staining technique was 82.0% (370/451) (P < 0.05). CONCLUSIONS: Magnifying FICE could show the mucosal microstructure and blood capillary form and it had a superiority of high coincident rate, high sensitivity and specificity when compared with ordinary magnifying colonoscopy and magnifying staining endoscopy. In addition, it was easy to operate and a biopsy could be taken from the target, so it has a satisfactory clinical practical value.

Embolization combined with endoscopic variceal ligation for the treatment of esophagogastric variceal bleeding in patients with cirrhosis.

BACKGROUND: Esophagogastric variceal bleeding caused by cirrhosis is a kind of emergent condition in the clinic. This study was conducted to explore the therapeutic effect and superiority of embolization of gastric fundus varices combined with endoscopic variceal ligation (EVL) of esophageal varices for the treatment of patients with esophagogastric variceal bleeding caused by cirrhosis. METHODS: Totally 172 patients were diagnosed by endoscopic examination within 24 hours of hospitalization with active gastric fundus variceal bleeding and grade II above esophageal varices caused by cirrhosis. Other causes leading to upper digestive tract bleeding were excluded. Patients were randomly divided into a control group (n = 82) and a therapy group (n = 90) following a random number table method. For the former, embolization for gastric fundus varices was performed, then an EVL for esophageal varices was performed 2 months later. For the therapy group, embolization for gastric fundus varices and EVL for esophageal varices were performed at the same time. RESULTS: The rate of emergency hemostasis in the therapy group was 100.0%, higher than that in the control group (87.8%, P < 0.05). The rate of early rebleeding in the therapy group was 6.7% while the rate in the control group was 23.6% (P < 0.05). No complications related to treatment occurred in both groups. CONCLUSION: Embolization for gastric fundus varices combined with EVL for esophageal varices is a safe and effective method for the treatment of patients with esophagogastric variceal bleeding caused by cirrhosis.

Endoscopic therapy for gastric stromal tumors originating from the muscularis propria.

AIM: To explore endoscopic therapy methods for gastric stromal tumors originating from the muscularis propria. METHODS: For 69 cases diagnosed as gastric stromal tumors originating from the muscularis propria, three types of endoscopic therapy were selected, based on the size of the tumor. These methods included endoscopic ligation and resection (ELR), endoscopic submucosal excavation (ESE) and endoscopic full-thickness resection (EFR). The wound surface and the perforation of the gastric wall were closed with metal clips. Immunohistostaining for CD34, CD117, Dog-1, S-100 and smooth muscle actin (SMA) was performed on the resected tumors. RESULTS: A total of 38 cases in which the tumor size was less than 1.2 cm were treated with ELR; three cases were complicated by perforation, and the perforations were closed with metal clips. Additionally, 18 cases in which the tumor size was more than 1.5 cm were treated with ESE, and no perforation occurred. Finally, 13 cases in which the tumor size was more than 2.0 cm were treated with EFR; all of the cases were complicated by artificial perforation, and all of the perforations were closed with metal clips. All of the 69 cases recovered with medical treatment, and none required surgical operation. Immunohistostaining demonstrated that among all of the 69 gastric stromal tumors diagnosed by gastroscopy, 12 cases were gastric leiomyomas (SMA-positive), and the other 57 cases were gastric stromal tumors. CONCLUSION: Gastric stromal tumors originating from the muscularis propria can be treated successfully with endoscopic techniques, which could replace certain surgical operations and should be considered for further application.

MicroRNA expression profile analysis reveals diagnostic biomarker for human prostate cancer.

Prostate cancer is a highly prevalent disease in older men of the western world. MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression via posttranscriptional inhibition of protein synthesis. To identify the diagnostic potential of miRNAs in prostate cancer, we downloaded the miRNA expression profile of prostate cancer from the GEO database and analysed the differentially expressed miRNAs (DE-miRNAs) in prostate cancerous tissue compared to non-cancerous tissue. Then, the targets of these DE-miRNAs were extracted from the database and mapped to the STRING and KEGG databases for network construction and pathway enrichment analysis. We identified a total of 16 miRNAs that showed a significant differential expression in cancer samples. A total of 9 target genes corresponding to 3 DE-miRNAs were obtained. After network and pathway enrichment analysis, we finally demonstrated that miR-20 appears to play an important role in the regulation of prostate cancer onset. MiR-20 as single biomarker or in combination could be useful in the diagnosis of prostate cancer. We anticipate our study could provide the groundwork for further experiments.

Efficacy of endoscopic therapy for gastrointestinal bleeding from Dieulafoy's lesion.

AIM: To investigate the endoscopic hemostasis for gastrointestinal bleeding due to Dieulafoy's lesion. METHODS: One hundred and seven patients with gastrointestinal bleeding due to Dieulafoy's lesion were treated with three endoscopic hemostasis methods: aethoxysklerol injection (46 cases), endoscopic hemoclip hemostasis (31 cases), and a combination of hemoclip hemostasis with aethoxysklerol injection (30 cases). RESULTS: The rates of successful hemostasis using the three methods were 71.7% (33/46), 77.4% (24/31) and 96.7% (29/30), respectively, with significant differences between the methods (P < 0.05). Among those who had unsuccessful treatment with aethoxysklerol injection, 13 were treated with hemoclip hemostasis and 4 underwent surgical operation; 9 cases were successful in the injection therapy. Among the cases with unsuccessful treatment with hemoclip hemostasis, 7 were treated with injection of aethoxysklerol and 3 cases underwent surgical operation; 4 cases were successful in the treatment with hemoclip hemostasis. Only 1 case had unsuccessful treatment with a combined therapy of hemoclip hemostasis and aethoxysklerol injection, and surgery was then performed. No serious complications of perforation occurred in the patients whose bleeding was treated with the endoscopic hemostasis, and no re-bleeding was found during a 1-year follow-up. CONCLUSION: The combined therapy of hemoclip hemostasis with aethoxysklerol injection is the most effective method for gastrointestinal bleeding due to Dieulafoy's lesion.

Application of endoscopic retrograde cholangiopancreatography in biliary-pancreatic diseases.

BACKGROUND: Endoscopic retrograde cholangiopancreatography (ERCP) has been used increasingly for the treatment of choledocholithiasis, gallstone pancreatitis, and benign or malignant bile duct or pancreatic duct stenosis. The purpose of this study was to evaluate ERCP for the diagnosis and therapy of biliary-pancreatic diseases. METHODS: A total of 2075 patients who underwent diagnostic and therapeutic ERCP from June 2001 to March 2009 were analyzed retrospectively. Achievement and complication rates were calculated, and the therapeutic effect was observed. RESULTS: In all the 64 cases who underwent diagnostic ERCP, the procedure was successful, in 2011 cases therapeutic ERCP was performed, and the success rate was 94.6%. In the therapeutic ERCP cases, 1434 (93.0%) were successful among the 1542 cases of choledocholithiasis, and 422 (90.0%) of the 469 cases with benign or malignant bile duct or pancreatic duct stenosis, or acute obstructive suppurative cholangitis with stent placement or endoscopic nasobiliary drainage were successful. Fifty-nine (90.8%) cases of the 65 who underwent a pre-cut for pancreatic sphincterotomy were successful. Complication rate was 5.1% and the most frequent complication was acute pancreatitis (4.4%). CONCLUSIONS: ERCP is one of the major diagnostic and therapeutic methods for biliary-pancreatic disease. Therapeutic ERCP is a minimally invasive, safe and effective treatment method for various biliary-pancreatic diseases.

Narrow-band imaging in the diagnosis of early esophageal cancer and precancerous lesions.

BACKGROUND: In the recent years, the incidence of esophageal cancer in China has increased. The key point for raising the survival rate is the diagnosis and treatment at an early stage. Narrow-band imaging (NBI) can enhance the contrast of the mucous membrane of the esophagus without staining. This study aimed to explore the value of NBI in the diagnosis of early esophageal cancer and precancerous lesions. METHODS: The esophagus was examined with ordinary endoscopy and NBI endoscopy. Pit patterns and blood capillary forms were examined with routine magnifying endoscopy and NBI endoscopy. Finally, a 1.2% Lugoul's iodine solution was used to stain the esophageal mucosal surface and a biopsy was taken at all the sites where NBI or iodine staining was positive. NBI and iodine staining scales were compared with pathologic diagnosis, which was considered as the gold standard. RESULTS: A total of 90 cases (138 lesions in total) were diagnosed as early esophageal cancer or precancerous lesions; 104 lesions (75.4%) were detected with ordinary endoscopy, 120 lesions (87.0%) were detected with NBI endoscopy, and 138 lesions (100%) were detected with iodine staining. The lesion detection rate of NBI was significantly lower than that of iodine staining (chi(2) = 17.176, P < 0.01). However, there was no significant difference between NBI and iodine staining for the diagnosis of high grade intraepithelial neoplasia (chi(2) = 1.362, P > 0.05), while the detection rate of NBI was significantly lower than that of iodine staining for the diagnosis of low grade intraepithelial neoplasia (chi(2) = 13.388, P < 0.01). The pit pattern and blood capillary form of early esophageal cancer and precancerous lesions could be demonstrated clearer with NBI than with ordinary endoscopy. CONCLUSIONS: NBI can enhance the contrast of the mucous membrane of the esophagus without staining. The combination of NBI and iodine staining can raise the diagnostic rate of early esophageal cancer and precancerous lesions.