Objective To establish an easy, rapid and sensitive method for determination of micro 2,4-dinitrotoluene in waste water. Methods Based on the fact that micro 2,4-dinitrotoluene could inhibit the discoloring oxidation reaction of methylene blue by KIO4 in the HC1 solution, micro 2,4-dinitrotoluene was determined using kinetic spectrophotometric method. Results A new kinetic spectrophotometric method for determination of micro 2,4-dinitrotoluene was established. The linear range of the method was 0.20?10~(-5)-1.8?10~(-5) mg/L, the detection limit was 1.7xlO'7 mg/L. When the method was applied to the determination of the 2,4-dinitrotoluene in waste water, its recovery rates in standard addition method were 95.6%-99.1% and the RSDs were 1.5%-2.6%. Conclusion The method was easier to operate and more rapid, which was suitable for the determination of micro 2,4-dinitrtoluene in wastewater.

Objective To investigate the expression of H19 imprinted genes in the villus tissues of women with spontaneous abortion.Methods Reverse transcription-polymerse chain reaction method(RT-PCR) was employed to detect H19 imprinted genes allele-specific expression levels of the villus tissues extracted from 45 cases of spontaneous abortion and 30 cases of normal pregnancy.Results Nineteen cases of 21 heterozygous cases in spontaneous abortion samples showed billelic expression of H19(19/21),whereas no billelic expression was found in the normal pregnancy samples(0/13)(P

Objective To reprogram the adipose-derived stem cells of hepatic cancer patients into induced pluripotent stem cells (iPS cells). Methods Retrovirue-based pMXs containing Oct4, Sox2, Klf4 and c-Myc were prepared and were used to infect adipose-derived stem cells; the infected cells were then cultured. Alkaline phosphatase staining, quantitative-PCR and immunofluorescence assay were used to identify the ES-cell-like colonies. Results The ES-cell-like colonies expressed Alkaline phosphatase. Quantitative-PCR showed that ES-cell-like cells expressed the pluripotency genes of ES cells. Furthermore, immunofluorescence confirmed the expression of Oct4 and Sox2 in ES-cell-like cells. Conclusion Adipose-derived stem cells of hepatic cancer patients can be efficiently reprogrammed into iPS cells, and they can serve as the feeder layers of their own iPS cells. These findings may pave a way for improving efficiency of reprogramming adult cells and establishing hepatic cancer model for inducing pluripotent stem cells.

OBJECTIVETo explore the clinical efficacy of acupoint catgut embedding therapy on chronic pelvic cavity pain syndrome differentiated as kidney deficiency and stagnation of damp heat, and explore the impacts on plasma P substance (SP), plasma beta-endorphin (β-EP).

METHODSOne hundred and eighty cases were randomly divided into a catgut embedding group (90 cases) and a western medication group (90 cases). In the western medication group, tamsulosin capsules 0. 2 mg were prescribed for oral administration, once a day; indometacin sustained release tablets, 25 mg, three times a day. Totally, the oral administration for 8 weeks was required. In the catgut embedding group, the acupoint catgut embedding therapy was applied to Qugu (CV 2), Shenshu (BL 23), Zhibian (BL 54), Huiyin (CV 1) and Sanyinjiao (SP 6), once every two weeks; the treatment of 4 weeks made one session, and two sessions were required. Before and after treatment, TCM symptom score, NIH-CPSI (the National Institute of Health Chronic Prostatitis Symptom Index) score, lecithin body numbers in prostatic fluid, score in SAS (self-rating anxiety scale), score in SDS (self-rating depression scale), the levels of SP and β-EP, etc. were observed in the two groups, and the clinical efficacy was assessed in the two groups.

RESULTS(1) Ten cases were dropped in either group. The total effective rate was 91. 25% (73/80) in the catgut embedding group, higher than 78. 75% (63/80) in the western medication group (P<0. 05). (2) After treatment, TCM symptom score, total score in NIH-CPSI, pain score and the scores in SAS and SDS were all reduced as compared with those before treatment in the two groups (all P<0. 05). After treatment, TCM symptom score, total score and pain score in NIH-CPSI, and the scores in SAS and SDS in the catgut embedding group were both lower than those in the western medication group (all P<0. 05). (3) After treatment, the lecithin body numbers were both increased as compared with those before treatment in the two groups (both P<0. 05), and the result in the catgut embedding group was higher than that in the western medication group (P<0. 05). (4)After treatment, the SP level was lower than that before treatment in the two groups (both P<0. 05); the level of p-EP was increased as compared with that before treatment (both P<0. 05). The SP level in the catgut embedding group was lower than that in the western medication group (P<0. 05); the level of β-EP was higher than that in the western medication group (P<0. 05).

CONCLUSIONThe acupoint catgut embedding therapy apparently relieves the clinical symptoms of chronic pelvic cavity pain syndrome differentiated as kidney deficiency and stagnation of damp heat as well as the condition of anxiety and depression, increases lecithin body numbers in prostatic fluid and β-EP level and reduces SP level.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Catgut ; utilization ; Chronic Disease ; therapy ; Humans ; Male ; Pelvic Pain ; etiology ; metabolism ; therapy ; Prostatic Diseases ; complications ; metabolism ; Young Adult ; beta-Endorphin ; metabolism

Objective: To explore the alternatives for T-tube placement in surgical treatment of bile duct stones.Methods: Fourty-nine patients with bile duct stones have been performed retrograde transhepatic biliary drainages(RTBD)since 1991.Among them,37 cases were placed the drainage tubes through the right hepatic lobes by retrograde punctures;the other 12 cases with intrahepatic stones were placed the drainage tubes through the intrahepatic bile duct stumps following partial hepatectomies.All the CBD exploration incisions were primarily closed.Results: (1)RTBD by retrograde puncture:Two postoperative complications(5.4%,pneumothorax and biliary hemorrhage)occured.Both cases were cured by medical therapy.The drainage tubes were removed on the 8th postoperative day.(2)RTBD through intrahepatic ducts:Four cases had residual stones(30%)which had been cleared up by combination of extraction and intraluminal lithotripsy.Nineteen cases(38.8%)had the gallbladders preserved.Postoperative cholangiographies showed that all the gallbladders were smooth and well-filled and the extrahepatic bile ducts were in normal shapes.Extrahepatic bile ducts and all the preserved gallbladders did not show any deformity and stone formation by ultrasonography during 6 months to 9 years'following-up.Conclusion: RTBD by retrograde puncture has the advantage of shorter period of bearing drainage tubes;RTBD through intrahepatic duct stump after partial hepatectomy makes it easier to extract postoperative residual stones.Both methods have no negative impacts on the anatomies of both the extrahepatic bile duct and the gallbladder.

Objective:To explore the application value of MRI texture analysis in combination with imaging features to predict the WHO/International Society of Urological Pathology (ISUP) nuclear grading in pre-operative patients with clear cell renal carcinoma (ccRCC).Methods:MRI images of 78 patients diagnosed as ccRCC by surgical pathology from July 2016 to July 2020 in First Affiliated Hospital of Fujian Medical University were retrospectively analyzed. According to the WHO/ISUP grading system, the patients were divided into low grade group (49 cases, grade Ⅰ in 2 cases and grade Ⅱ in 47 cases) and high grade group (29 cases, grade Ⅲ in 25 cases and grade Ⅳ in 4 cases), and then were assigned to training set ( n= 63) and validation set ( n=15) in a ratio of 7∶3 using random indicator method. MRI radiological features were evaluated and MRI imaging texture features were extracted. The largest-diameter slice of lesion on cross-sectional images was selected and ROIs were drawn on T 2WI and corticomedullary phase (CMP) images, respectively. Quantitative texture analysis software MaZda was used to extract texture features, including gray-scale histogram, co-occurrence matrix, run-length matrix, gradient, autoregressive model and wavelet transform. The extracted texture features were preliminarily selected by the combination of Fisher, probability of classification errorand average correlation coefficient, and interaction information, and then the reduced texture parameters or imaging features were tested by the independent sample t test, Mann-Whitney U test or χ 2 test. Parameters with statistically significant differences were used to construct a multi-factors binary logistic regression model and the ROC curve was used to analyze its effectiveness in predicting high grade ccRCC. Results:In training set, there were significant differences intumor length, shape and margin, enhancement degree of CMP, vein thrombosis and 47 texture features between the low and high grade ccRCC groups. In the training set, 7 multi-factors binary logistic regression model were constructed, including radiological features model (M1), T 2WI texture features model (M2), CMP image texture features model (M3) and combination radiological features of T 2WI texture features model (M4), combination radiological features of CMP images texture features model (M5), combination T 2WI texture features of CMP images texture features model (M6) and combination of all features model (M7). The area under ROC curve of M7 in predicting nuclear grading of ccRCC was the largest, which were 0.901 (95% CI 0.828-0.974) and 0.820 (95% CI 0.564-0.974) in the training set and validation set, respectively. Conclusion:MRI texture analysis combined with imaging features is hopeful to be an effective preoperative noninvasive method in predicting WHO/ISUP grading of ccRCC.

Objective: To observe the preventive effect of alkaline drinking water on hyperuricemia in mice. Methods: Sixty male SPF Kunming mice were randomly divided into six groups: pH 7.3, pH 8.0, pH 9.3 intervention groups, in which the mice were given water with pH values of 7.3±0.5, 8.0±0.5 and 9.3±0.6, respectively; the control group, model group and positive drug group ( with 2 g/L allopurinol ) were given double distilled water. Except for the control group, the mice in each group were given yeast by gavage (1.5 g/mL) for 13 days. On the 14th day, the mice were injected with 300 mg/kg potassium oxyzinate by intraperitoneal injection, and then fasted for 1 day. On the 16th day, serum uric acid, creatinine and urea nitrogen were detected, and renal tissues were stained to observe the morphology.The expression levels of neutrophil gelatinase-associated lipocalin ( NGAL ), tissue inhibitor of metalloproteinase 1( TIMP1 ), organic anion transporter 1 ( OAT1 ) and urate transporter 1 ( URAL-1 ) in renal tissues were determined bywestern blotting. The mRNA expression levels of URAL-1 and OAT1 were detected by real-time fluorescent quantita⁃tive polymerase chain reaction. Results: The level of serum uric acid was higher in the model group than in the control group and in the pH 9.3 intervention group (both P<0.05). The number and area of renal tubular lesions were less in the pH 9.3 intervention group than in the model group (all P<0.05). The relative expression levels of NGAL and URAT-1 proteins were lower in the pH 9.3 intervention group than in the model group, and the relative expression level of OAT1 protein was higher in the pH 9.3 intervention group than in the model group ( all P<0.05). The relativeexpression level of URAT-1 mRNA was lower in the pH 9.3 intervention group than in the model group, and the rela⁃tive expression level of OAT1 mRNA was higher in the pH 9.3 intervention group than in the model group ( all P<0.05 ). Conclusion Alkaline drinking water with pH value of 9.3±0.6 can effectively prevent hyperuricemia and acute kidney injury in mice.

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

Objective To research the influenza virus infection on rat vascular smooth cells number,proliferation,apoptosis,the amount of IL-6,sFas,platelet-derived growth factor(PDGF) and the mechanism of atherosclerosis.Methods Flow cytometry,enzyme-linked immunosorbent assay and cell count experiments were used to detect these indicators at 0 h,6 h,12 h,24 h,48 h.Results After influenza virus infected at 0 h,proliferation,apoptosis condition were 10.39％,0.44％,respectively; at 6 h,proliferation,apoptosis respectively increased to 12.68％,0.73％ ; proliferation reached the peak at 12 h (18.01％),instead apoptosis decreased to 0.14％ ; at 24 h,proliferation decreased to 12.89％ and apoptosis markedly increased to 1.09％ ; at 48 h,proliferation further reduced to 7.07％ and apoptosis reached the peak(4.61％).The number of cells and the cytokine secretion were statistically significant to control group (P＜0.05).Conclusion Influenza virus infection might lead to change of cell proliferation and apoptosis and involve the atherosclerosis form and development,and cytokines played an important role in them.

Objective To establish a high-throughput evaluation model for anaphylactic reactions; To screen and identify potential anaphylactogens from TCM monomeric compounds.MethodsCell model of stably expressed MrgX2 was established. Recombinate plasmid pmCherry-C1-MrgX2 was transfected to HEK293 to establish cell line for screening model. MrgX2 agonist and antagonist were used to identify the validation and stability of the cell line. A small library consisting of 180 compounds was profiled by using a cell-based calcium mobilization assay to find novel compounds targeting the MrgX2 receptor. EC50 test, IC50 test, specificity validation and cytotoxicity evaluation were carried out to detect the function of the positive agonist.ResultsThe EC50 of C48/80 to MrgX2 model was 2.7 μg/mL and the IC50 of 2-APB (evoked by 10 μg/mL C48/80) was 46.29 μmol/L. The first generation cell model of MrgX2 was similar to the 20th generation, and the Z factor of MrgX2 cell model was 0.78. In the primary screening for agonist, isoliensinine was identified as a novel agonist targeting receptor MrgX2 with an EC50 of 4.5 μmol/L and IC50 of39.47 μmol/L. Moreover, isoliensinine was validated to activate MrgX2 receptor specifically without cytotoxicity. Conclusion A high-throughput evaluation method for anaphylactic reactions can be established in vitro through calcium mobilization assay. A potential anaphylactogen isoliensinine is identified and validated.