OBJECTIVETo establish the best hydrolytic conditions from phorbol esters.

METHODThe orthogonal experiment was used to optimize 4 factors, which were reaction time, ratio of solid-to-liquid, hydrolytic times, and temperature. Diamonsil C18 column (4. 6 mm x 250 mm, 5 microm) was used and the mobile phase was consisted of acetonitrile and water for HPLC detection. The detection wavelength was set at 234 nm, the flow rate was 1 mL x min(-1), and the column temperature was 25 degrees C.

RESULTThe optimum conditions were 10 h of reaction time, 1:6 of solid-to-liquid (BaOH/MeOH) ratio, 25 degrees C of temperature, and one time of hydrolysis. There was a good linear relationship of phorbol in the range of 4.28-107 mg x L(-1) (r = 0.999 9), and the average recovery was 97.89%, with RSD 0.78%.

CONCLUSIONThe method is steady, reliable and reproducible, and it provides a mean for future study.

Chromatography, High Pressure Liquid ; Hydrolysis ; Phorbol Esters ; analysis ; chemistry

Peripheral blood CD_5~+B cell was detected by flow cytometry in patients with Graves′ disease (GD) before and after treatment. As compared with normal controls, peripheral blood CD_5~+B cells in a group of 43 patients with GD showed a significant increase in number [(17.0+5.1)% vs (39.5+12.4)%, P

OBJECTIVETo generate hepatitis C virus pseudo-particles (HCVpp) containing the complete E1-E2 envelope glycoprotein, in order to establish a HCVpp database covering the six major genotypes of HCV (1b, 2a, 3b, 4, 5, and 6) and to develop a simple and effective method for detection of neutralizing antibodies in HCV patients.

METHODSHCVpp were generated for the six genotypes by co-transfecting 293T cells with a plasmid expressing the respective E1-E2 (p HR, CMVA 8.2 construct) and a MLV-GFP plasmid. Titration of each HCVpp was carried out by p24 ELISA. Infectivity of each HCVpp was assessed by mixing the harvested supernatant of producer cells with sera from HCV patients, adding the mixture to Huh-7 cells, and detecting the subsequent titers of neutralizing antibodies against HCVpp.

RESULTSAll six types of HCVpp were able to infect Huh-7 cells in vitro. For healthy HCV carriers, only two genotypes of HCVpp (1b and 2a) produced neutralizing antibody titers more than 1:40. For cured HCV patients, only the 1b genotype produced neutralizing antibody titers more than 1:40. One patient showed titer of 1:200 for genotype 4. A healthy spouse of a chronic hepatitis C patient showed titers more than 1:40 for four genotypes of HCVpp (3a, 4, 5, 6).

CONCLUSIONWe generated six different genotypes of HCVpp successfully, established the in vitro neutralizing antibody detection method, and provided an effective model for screening antiviral drugs.

Adolescent ; Adult ; Antibodies, Neutralizing ; blood ; Antibodies, Viral ; blood ; Female ; Genotype ; Hepacivirus ; classification ; Hepatitis C ; blood ; immunology ; Humans ; Male ; Middle Aged ; RNA, Viral ; blood ; Viral Envelope Proteins ; immunology ; Young Adult

OBJECTIVETo investigate the ideal digestive tract reconstruction methods among three different surgical methods after radical gastrectomy of gastric cancer patients.

METHODSA total of 123 patients who received elective radical gastrectomy for gastric cancer from February 2010 to August 2011 were prospectively enrolled and randomly divided into radical proximal gastrectomy and jejunal interposition group, radical proximal gastrectomy and esophageal with the posterior of residual-stomach group, and radical total gastrectomy and Roux-en-Y esophagojejunostomy group. Patients were followed up for 12 months. Symptoms of reflux esophagitis were observed, gastric emptying tests were done, liver and kidney function was also monitored. The quality of life was documented before operation, and one and twelve months after operation.

RESULTSNo significant differences were found among these three groups in the pH value of lower part of esophagus, the blood regular test results and the functional parameters of kidney and liver before and after operation(all P>0.05). Symptoms of reflux esophagitis was reported in 1(2.4%) patients in the jejunal interposition group, 10(24.4%) in esophageal with the posterial of residual-stomach group, and 7(17.1%) in the Roux-en-Y esophagojejunostomy group(P=0.017). There was 1(2.4%), 10(17.1%), and 8(19.5%) patients presented reflux of barium meal in these three groups, respectively (P=0.046). There were no statistically significant difference in PH at the distal esophagus(6.9±0.2 vs. 6.8±0.1 vs. 6.9±0.1, P=0.196). The quality of life was significantly improved one year after surgery in terms of general status, physical function, emotional function, fatigue, nausea/vomiting, pain, constipation, and diarrhea (all P<0.05), with the jejunal interposition superior than the other two methods.

CONCLUSIONThree methods of digestive tract reconstruction in radical gastrectomy of gastric cancer patients can improve the health status and the quality of life in gastric cancer patients. Radical proximal gastrectomy and jejunal interposition is the preferred method.

Anastomosis, Roux-en-Y ; Anastomosis, Surgical ; Digestive System Surgical Procedures ; Esophagus ; Gastrectomy ; Gastric Emptying ; Gastric Stump ; Humans ; Jejunum ; Quality of Life ; Reconstructive Surgical Procedures ; methods ; Stomach Neoplasms ; surgery

Abdominal Wall ; pathology ; Humans ; Male ; Mesentery ; pathology ; Middle Aged ; Ossification, Heterotopic ; etiology ; Postoperative Complications ; etiology ; Tomography, X-Ray Computed

OBJECTIVETo establish an animal model of lung injury in SD rats using intratracheal instillation of unknown polymer and to provide the base for exploring the molecular mechanism of lung tissue injury induced by occupational exposure.

METHODSOne hundred forty SD rats were randomly divided into seven groups, including the control group 1 which was exposed to normal solution, the control group 2 which was not exposed to any one and five treatment groups which were exposed to 1 ml unknown polymer (0.5 ml for each lung) at the doses of 40, 30, 20, 10 and 5 mg/ml, respectively by intratracheal instillation. The rats were sacrificed on the 1st, 3rd, 7th, 10th, 14th, 21th and 28th day after exposure, then the lung tissues were examined pathologically and the blood bio-chemical analysis was conducted.

RESULTSThe results of blood biochemical analysis indicated that ALT and AST levels in rats exposed to 30 and 40 mg/ml unknown polymer were significantly higher than those in control groups. Intratracheal instillation of unknown polymer can causes PLF in experimental animals on the 14th days after exposure. The results of pathological examination exhibited that the lung tissue injury in rats exposed to unknown polymer for 14 days or more was found and the dose-effect relationship was observed.

CONCLUSIONAn animal model of lung injury in SD rats induced by unknown polymer with intratracheal instillation was established successfully. The results of pathological examination showed that the types of rat lung injury were similar to the clinical lung injury after exposure to unknown polymer, which provided a base for studying the mechanism of lung injury caused by occupational exposure to unknown polymer.

Animals ; Disease Models, Animal ; Female ; Lung ; drug effects ; pathology ; Lung Injury ; chemically induced ; Male ; Polymers ; toxicity ; Rats ; Rats, Sprague-Dawley

Objective To investigate the diagnostic value of pleural effusion cell block (PECB) immunohistochemical examination in patients with suspected lung cancer.Methods Eighty-six patients with suspected lung cancer and pleural effusion were selected from December 2014 to December 2016 in the First People's Hospital of Zaoyang City.Liquid based cytology (LBC) smear was used for the diagnosis of cell morphology.Immunohistochemical detection of cell blocks was carried out,including CK7,CK5/6,p53,thyroid transcription factor-1 (TTF-1),calretinin,CD56,estrogen receptor(ER),progesterone receptor(PR),human epidermal growth factor receptor-2 (HER-2),carcinoembryonic antigen (CEA),cancer antigen 125 (CA 125)and calretinin.Immunologic marker staining was further carried out according to the cell morphology and clinical data.Results LBC smear test showed that there were 61 cases of metastatic adenocarcinoma,11 cases of metastatic poorly differentiated carcinoma,3 cases of metastatic small cell carcinoma,2 cases of metastatic large cell carcinoma,3 cases of metastatic squamous cell carcinoma,2 cases of malignant tumor and 4 cases of suspicious malignant tumor in the 86 cases.Immunohistochemical results showed that there were 69 cases of primary lung metastases (including 61 cases of pulmonary adenocarcinoma,2 cases of lung squamous cell carcinoma,one case of lung adenosquamous carcinoma and 5 cases of small cell lung cancer),5 cases of metastatic adenocarcinoma of extrapulmonary origin (including 2 cases of breast cancer,one case of prostatic cancer,one case of ovarian cancer and one case of pancreatic cancer),3 cases of metastatic squamous cell carcinoma(including 2 cases of esophageal carcinoma and one case of cervical cancer),2 cases of metastatic adenosquamous carcinoma(including one case of adenosquamous carcinoma of cervix and one case of adenosquamous carcinoma of rectum),2 cases of metastatic large cell carcinoma,3 cases of large cell neuroendocrine carcinoma,1 case of mesothelioma,and 1 case without definite diagnosis in the 86 cases.In the 69 cases of primary lung tumor metastasis,there were 63 cases with positive expression of TTF-1 and CK7 protein,2 cases with positive expression of CK20 protein and 2 cases with positive expression of p53 protein.The expression of CK5/6,CD56 and calretinin protein was negative in the 69 cases.In the 5 cases of metastatic adenocarcinoma of extrapulmonary origin,there was 1 case of breast carcinoma with positive expression of ER,PR and HER-2 protein,1 cases of breast carcinoma with positive expression of ER and PR protein,1 cases of prostatic cancer with positive expression of CK5/6 protein,1 cases of ovarian cancer with positive expression of ER protein and 1 cases of pancreatic cancer with positive expression of CEA and CA125 protein.In the 3 cases of metastatic squamous cell carcinoma,there were 2 cases of esophageal cancer with positive expression of CEA and p53 protein and 1 case of cervical cancer with positive expression of ER protein.In the 2 metastatic adenosquamous carcinoma,there was 1 case of adenosquamous carcinoma of cervix with positive expression of PR and TTF-1 protein and 1 case of adenosquamous carcinoma of rectum with positive expression of CK7 and CEA protein.Conclusion PECB immunohistochemical examination can accurately diagnose primary focus and subtype lung cancer,and further help to determine the location of the exogenous tumor.

Objective:To investigate the application value of molecular detection in the differential diagnosis of ovarian adult granulosa cell tumors (AGCT) by analyzing FOXL2, AKT1 and DICER1 mutations in these tumors.Methods:A total of 48 cases of ovarian sex cord-stromal tumor (SCST) were selected from July 2012 to June 2019 in Beijing Obstetrics and Gynecology Hospital, including 21 adult granulosa cell tumors (AGCT), 15 fibromas/fibrothecomas, 8 Sertoli-Leydig cell tumors (SLCT) and 4 other types of ovarian SCST. Genomic DNA was extracted from the formalin-fixed paraffin-embedded tissue sections. Polymerase chain reaction amplification for FOXL2, AKT1 and DICER1 genes was performed, followed by sequencing using capillary electrophoresis. Fisher exact test was used to compare the prevalence difference of FOXL2, AKT1 and DICER1 mutations among the groups. P<0.05 was considered significant. Results:Eighteen of the 21 (85.7%) AGCT harbored FOXL2 mutation. Compared with other SCST (13.0%, 3 of 23; including fibromas/fibrothecomas and SLCT), FOXL2 mutation was significantly higher in AGCT ( P<0.001). In addition, FOXL2 mutation was also detected in one fibrothecoma, two SLCT and two gynandroblastomas. DICER1 mutation was identified in four of eight SLCT, and these cases were moderately to poorly differentiated. FOXL2 mutation was found in one SLCT with DICER1 mutation. There was no DICER1 mutation in other ovarian SCST. No AKT1 mutation was detected in all the patients. Conclusions:FOXL2 mutation is a highly specific biomarker for adult AGCT and may be helpful to resolve problematic cases. Diagnosis should also be taken into consideration of the clinical and histological features as FOXL2 mutation is also found in other SCST. The detection of DICER1 mutation is helpful for the differential diagnosis of ovarian SLCT. Synchronous DICER1 and FOXL2 mutation in the SLCT has been observed, and its significance needs to be further studied.

OBJECTIVETo study the effects of enhanced external counterpulsation (EECP) on the vascular morphology, and endothelial function using experimentally induced hypercholesterolemic pigs.

METHODSThirty five male pigs were randomly divided into three groups: 7 normal control animals, 11 hypercholesterolemic animals, and 17 hypercholesterolemic animals receiving EECP. Serum cholesterol was measured. The coronary arteries and aortas were sampled for histopathologic and ultrastructural examination. The NF-kappaB protein expression of porcine coronary arteries was investigated by immunofluorescence.

RESULTSCompared with the normal controls, serum cholesterol levels were significantly higher in the hypercholesterolemic animals with or without EECP. The plaque/intimal area ratio of the aorta decreased significantly in animals receiving EECP [(3.33 +/- 2.40)%, versus (12.03 +/- 7.12)% in those without EECP, P < 0.05]. Lipid deposition, endothelial damage and proliferation of smooth muscle cells were less severe in animals receiving EECP than those not. Moreover, activation and expression of NF-kappaB also decreased significantly (P < 0.05) in animals receiving EECP.

CONCLUSIONSEECP improves the morphology and function of vascular endothelium, and retards the development and progression of atherosclerosis, likely through the inhibition of NF-kappaB signaling pathway.

Animals ; Aorta, Abdominal ; metabolism ; pathology ; ultrastructure ; Atherosclerosis ; blood ; metabolism ; pathology ; Cholesterol ; blood ; Coronary Vessels ; metabolism ; pathology ; ultrastructure ; Counterpulsation ; methods ; Endothelial Cells ; metabolism ; pathology ; Hypercholesterolemia ; blood ; metabolism ; pathology ; Lipoproteins, LDL ; blood ; Male ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Muscle, Smooth, Vascular ; metabolism ; pathology ; NF-kappa B ; metabolism ; Random Allocation ; Swine

Purpose To investigate the clinical manifesta-tions and morphologic features of placental site nodules (PSNs), and its clinical significance. Methods Twenty patients diag-nosed as PSNs were collected, then a retrospective analysis was conducted, and the characteristics of clinical data and follow-up results were analyzed,including of clinical manifestations, ultra-sonographic evaluation, morphologic and immunohistochemical features. Results The age of patients ranged from 25 to 41 years (32. 48 ± 4. 77 years in average). Three fifths of patients had pregnancy history for at least two times and the interval time to the last pregnancy ranged from 5 to 37 months (15. 33 ± 8. 05 months on average). 15 (75% ) patients went to the hospital because of abnormal vaginal bleeding. In our study, most of the samples showed a membrane-like structure without definite nod-ule. Microscopically, single or multiple, well-circumscribed and oval small nodules were found in endometrial tissue. In most ca- ses, the hyalinization was generally uniform in the center of the nodules, more or less intermediate trophoblasts appeared on the edge of the nodules. Immunohistochemically, the strong diffuse expressed CK (AE1/AE3), CAM5. 2, EMA, GATA-3, Cyclin E and p63 were detected in most of all cases, and PLAP showed strong focal expression, α-inhibin and hPL showed faint focal expression, Ki-67 staining for proliferative index was less than 4% . Conclusion PSN is a benign lesion of the intermediate trophoblast at the chorionic leave. Some diseases including hya-linized decidua, epithelioid trophoblastic tumor, and squamous cell carcinoma with hyalinization need to be identified. Some im-munohistochemical markers may be certain helpful in distinguis-hing as necessary.