Excretion of three alkaloids from Simiao Pills in urine, feces, and bile between normal and type 2 diabetic rats / 中国中药杂志

This study investigated the differences in excretion kinetics of three alkaloids and their four metabolites from Simiao Pills in normal and type 2 diabetic rats. The diabetes model was established in rats by injection of streptozotocin, and the alkaloids in urine, feces, and bile of normal and diabetic rats were detected by LC-MS/MS to explore the effect of diabetes on alkaloid excretion of Simiao Pills. The results showed that 72 h after intragastric administration of the extract of Simiao Pills, feces were the main excretion route of alkaloids from Simiao Pills. The total excretion rates of magnoflorine and berberine in normal rats were 4.87% and 56.54%, which decreased to 2.35% and 35.53% in diabetic rats, which had statistical significance(P<0.05). The total excretion rates of phellodendrine, magnoflorine, and berberine in the urine of diabetic rats decreased significantly, which were 53.57%, 60.84%, and 52.78% of those in normal rats, respectively. After 12 h of intragastric administration, the excretion rate of berberine in the bile of diabetic rats increased significantly, which was 253.33% of that of normal rats. In the condition of diabetes, the excretion rate of berberine metabolite, thalifendine significantly decreased in urine and feces, but significantly increased in bile. The total excretion rates of jateorrhizine and palmatine in the urine increased significantly, and t_(1/2) and K_e changed significantly. The results showed that diabetes affected the in vivo process of alkaloids from Simiao Pills, reducing their excretion in the form of prototype drug, affecting the biotransformation of berberine, and ultimately increasing the exposure of alkaloids in vivo, which would be conducive to the hypoglycemic effect of alkaloids. This study provides references for the clinical application and drug development of Simiao Pills in diabetes.

Study on effects of dioscin against depression and its mechanisms / 中国药理学通报

Aim To investigate the effects and possible molecular mechanisms of dioscin(DIO)against depression in mice.Methods Eighty mice were randomly divided into control group, DIO control group, model group, DIO groups(20, 40 and 80 mg·kg-1 DIO)and FLU group(20 mg·kg-1 fluoxetine).After establishing the depression model with chronic unpredictable moderate stress(CUMS)in mice, the corresponding drugs were administered by gavage continuously for four weeks in each group.The behavior of mice was tested, and the contents of corticosterone(CORT), brain-derived neurotrophic factor(BDNF), 5-hydroxytryptamine(5-HT), malondialdehyde(MDA), superoxide dismutase(SOD)and catalase(CAT)were evaluated by ELISA or enzyme labeling method.In addition, HE staining, Nissl staining and PET scanning were operated for the brain tissues.Western blot was used to detect the protein expressions.Results Compared with model group, DIO significantly improved the behaviors of depressed mice.And it reduced the contents of CORT in serum, increased BDNF and 5-HT in hippocampus.Meanwhile, DIO obviously reduced MDA in serum, increased SOD in serum and CAT in brain tissues.DIO improved the steatosis of brain tissues, disorder and looseness of neurons, and increased glucose metabolism in brain tissues of depressed mice.The molecular mechanism suggested that compared with model group, DIO significantly increased the protein level of UCP2 to adjust the levels of Nrf2, SOD2, GLUT1 and G6Pase.Conclusions DIO improves the depression symptoms of depressed mice, which should be through adjusting UCP2-mediated oxidative stress and glucose metabolism.

Clinical features and treatment efficiency of 6 children with Dent disease / 中华实用儿科临床杂志

Objective:To summarize the clinical features, genetic testing and treatment efficacy of 6 children with Dent disease.Methods:Six children diagnosed with Dent disease in Guangzhou Women and Children′s Medical Center from January 2014 to March 2019 were enrolled.Their medical history, clinical manifestations, laboratory results, genetic test results, and proteinuria level, calciuria level and renal function after medication were measured.Results:All patients were male, with the onset age ranged from 1 to 9 years old.They were followed up for 6 months to 4 years.All the children had low molecular weight proteinuria.Urine protein electrophoresis showed that the ratio of low molecular weight proteinuria in only 2 cases was more than 50%.Renal biopsy suggested that all cases were combined with glomerular lesions.Five cases had hypercalciuria.Under the microscope, there were 5 cases of hematuria.Two case had rickets, and there was no renal calcium deposition and hypophosphatemia.Five cases were detected with CLCN5 mutations, of which p. C160Yfs*49 and p. G523D were first reported.One case had an OCRL1 mutation.Patients were treated with Hydrochlorothiazide and angiotensin converting enzyme inhibitor (ACEI). The 24 h urinary calcium level after treatment was lower than that before treatment [0.40 (0.24, 0.43) mmol/kg vs.0.12 (0.11, 0.14) mmol/kg, U=2.00, P<0.01]. However, there was no significant decrease in the 24 h-urinary protein level before and after treatment [77.09 (62.41, 88.01) mg/kg vs.80.33 (66.03, 92.52) mg/kg, U=12.00, P>0.05]. Conclusions:Dent disease is mainly characterized by low molecular weight proteinuria, and some patients may not be associated with hypercalciuria.Gene tests help to identify the disease type.ACEI and Hydrochlorothiazide can reduce the urinary calcium level, but cannot improve the level of urinary protein.

Comparative analysis of pharmacokinetics and intestinal absorption of six alkaloids between Sanmiao Pills and Simiao Pills in rats / 中国中药杂志

The present study investigated the differences in pharmacokinetics and intestinal absorption of six alkaloids in Sanmiao Pills and Simiao Pills in rats and explored the different efficacies of the two formulae. After oral administration of Sanmiao Pills and Simiao Pills in rats, blood samples were collected at different time points. Samples were prepared for the determination of six alkaloids in plasma by UPLC-MS/MS. The chromatography was performed on an ACE Excel 3 C_(18 )column with acetonitrile-0.1% formic acid in water as the mobile phase for gradient elution. Analytes were detected in the positive ion mode. Plasma concentrations and pharmacokinetic parameters were calculated. Intestinal absorption of alkaloids was investigated by single-pass intestinal perfusion and absorption parameters of ingredients were calculated. The results showed that the UPLC-MS/MS method for simultaneous determination of concentrations of six alkaloids in plasma was developed and validated by methodological investigations, such as specificity, calibration curves, precision, accuracy, recovery, matrix effect, and stability. The results of the pharmacokinetic assay revealed that C_(max) and AUC values of phellodendrine, berberine, magnoflorine, berberrubine, and jatrorrhizine in Simiao Pills were significantly increased, and CL/F values were reduced as compared with those in Sanmiao Pills, which indicated the increase in plasma concentrations of alkaloids. The intestinal absorption parameters K_(a )and P_(eff) values of phellodendrine, berberine, and jatrorrhizine in Simiao Pills were higher than those in Sanmiao Pills. The intestinal absorption and plasma concentrations of alkaloids in Simiao Pills were significantly higher than those in Sanmiao Pills, suggesting that the composition of Simiao Pills was more conducive to the alkaloids into the blood to resist inflammation and lower uric acid.

Fever for 2 months and disturbance of consciousness for 1 week in a preschool-aged girl / 中国当代儿科杂志

Rapid-onset obesity with hypothalamic dysfunction, hypoventilation, and autonomic dysregulation syndrome (ROHHADS) is a rare multi-system disease, and delayed diagnosis and treatment may lead to catastrophic cardiopulmonary complications. As far as we know, no patient with ROHHADS has been reported in China, and this article reports a child with ROHHADS to improve the awareness of this disease among clinicians. A girl, aged 3 years, had the clinical manifestations of rapid weight gain, fever, disturbance of consciousness, and convulsion. The physical examination showed a body weight of 20 kg, somnolence, irregular breathing, and stiff neck. She had increased blood levels of prolactin and follicle-stimulating hormone and hyponatremia. The lumbar puncture showed an increased intracranial pressure. The brain MRI and magnetic resonance venography showed symmetrical lesions in the periventricular region and venous thrombosis in the right transverse sinus and the superior sagittal sinus. The sleep monitoring showed hypopnea. The girl was finally diagnosed with ROHHADS and intracranial venous thrombosis. She recovered after symptomatic treatment including decreasing intracranial pressure, anticoagulation, and respiratory support. The possibility of ROHHADS should be considered for patients with unexplained obesity, fever, and hypoventilation, with or without central nervous system symptoms. Early diagnosis and standardized follow-up can improve the prognosis of children with ROHHADS.

Clinical analysis of seven cases with primary hyperoxaluria type 1 in children / 中华儿科杂志

Objective@#To investigate the clinical, imaging and molecular characteristics of primary hyperoxaluria type 1 (PH1) in children and to sum up existing evidence for further understanding the phenotype-genotype correlation of infantile PH1.@*Methods@#This retrospective analysis was based on the medical records of children with PH1 diagnosed by gene test in the Department of Nephrology, Guangzhou Women and Children′s Medical Center from June 2016 to May 2019. Targeted exome sequencing was performed on tubular disease-related genes of the probands and Sanger sequencing was conducted to validate suspected pathogenic variants of family members. Logistic regression analysis of NC and CCr was adopted to show the relation between NC and renal function. The literature review was conducted, and the clinical, imaging and molecular biogenetic characteristics of the disease were analyzed and summarized.@*Results@#A total of 7 children from 6 families were enrolled. The median age of onset was 5 months. The median age of diagnosis was 8 months. Five cases had progressed to end-stage renal disease (ESRD), one case had chronic kidney disease (CKD) stage 1, and the other one had CKD stage 2. Four cases died, one case maintained on hemodialysis, and the other two non-dialysis cases were followed up. Among the 7 cases, 4 patients had infantile PH1, 1 patient had child and adolescent type, 1 patient had family type and the other one had unknown classification. There were two siblings (the younger brother had uremia and the sister had normal renal function) who had the delayed diagnosis for 5 and 3 years respectively. All patients in this cohort had proteinuria and microscopic hematuria, but no patients had gross hematuria. Three cases had hypercalciuria. Comprehensive diagnostic imaging evaluation include CT scan, MR scan, radiography and ultrasound led to the diagnosis of nephrocalcinosis (NC) in 5 cases, including 4 cases of simple NL and 1 case of NC with nephrolithiasis (NL), 1 case of multiple NL and 1 case of microcrystal deposition in renal medulla. However, only one case of NC was identified by ultrasound, the other 4 cases of NC were identified by radiograph examination. In the logistic regression analysis involving NC and creatinine clearnce rate (CCr), the results showed that NC was an independent risk factor for renal dysfunction (OR 2.5, 95%CI 0.7-1.2, P<0.05). All the 7 cases had AGXT gene variant, including homozygous variant in 4 cases and compound heterozygous variant in 3 cases. A total of 9 variant genotypes were found, and exon 6 variants were found in 4 children. Among them, there were 3 cases with c.679_680delAA. To our knowledge, both c.679_680delAA and c.190A>T in the cohort have not been reported previously.@*Conclusions@#Infantile PH1 is the most common type of PH1 in children, which progresses rapidly or even begins with renal failure, with poor prognosis. It is also highly heterogeneous in phenotype and genotype. NC is an independent risk factor leading to renal failure. Radiograph examination showed high specificity for the diagnosis of NC. At present, the misdiagnosis and delayed diagnosis of PH1 are still common in China. It is of great significance to carry out quantitative determination of uric oxalate in order to reduce the misdiagnosis rate and enhance follow-up technologies for evaluating the therapeutic effect.

Effect of Silencing and Overexpression of LNK Gene on STAT3 Expression in THP-1 Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of LNK gene silencing and overexpression on the expression of STAT3 gene in human monocytic leukemia cells (THP-1).@*METHODS@#THP-1 cells were cultured, and the lentivirus was used as a vector to silence and overexpres the LNK gene stably. After transfection for 72 hours, the GFP expression levels were observed by inverted fluorescence microscopy. The lentiviral transfection efficiencies were detected by flow cytometry. The effects of LNK silencing and overexpression were confirmed, and the expression of STAT3 mRNA was detected by RT-PCR. The protein levels of LNK and STAT3 were detected by Western blotting.@*RESULTS@#The GFP expression level of THP-1 cells reached more than 85% after transfection with lentivirus for 72 hours, and the transfection efficiency of cells was above 99%. mRNA expressions levels of LNK and STAT3 in LNK silencing group were signifycantly lower than those in control group, while LNK and STAT3 mRNA levels in the LNK overexpression group was significantly higher than those in control group (P＜0.05). The protein expression levels of LNK and STAT3 in LNK silencing group were significantly lower than those in control group, while that in LNK overexpression group was significantly higher than that in control group (P＜0.05).@*CONCLUSION@#The THP-1 cell line with LNK gene silencing and overexpression has been successfully established. The LNK gene silencing resulted in decrease of STAT3 expression; LNK gene overexpression and leads to inereases of STAT3 expression indicating that LNK participates in the regulation of STAT3.

Determinants of prognosis in Talaromyces marneffei infections with respiratory system lesions / 中华医学杂志(英文版)

BACKGROUND@#Little study has investigated the differences between Talatomyces marneffei (T. marneffei) respiratory infection and tuberculosis and the prognostic factors of such infection. This study investigated the characteristics and prognostic factors of T. marneffei infections with respiratory lesions and the causes of misdiagnosis.@*METHODS@#Clinical characteristics and prognoses of patients with T. marneffei infections with respiratory system lesion were investigated. T. marneffei diagnosis followed isolation from clinical specimens using standard culture, cytology, and histopathology. Survival curves were estimated by using Kaplan-Meier analysis, with log-rank test to compare differences in survival rates between groups. Univariate and multivariate Cox regression analyses were also performed to assess significant differences in clinical characteristics of overall survival.@*RESULTS@#Of 126 patients diagnosed with T. marneffei infections, 63 (50.0%) had T. marneffei respiratory system infections; 38.1% (24/63) were misdiagnosed as having tuberculosis. Human immunodeficiency virus (HIV) infection, CD4/CD8 105 days were potential risk factors for poor prognoses. Length of time from onset to confirmation of diagnosis persisted as an independent predictor of all-cause mortality in multivariate analysis (odds ratio: 0.083, 95.0% confidence interval: 0.021-0.326, P < 0.001). However, the size of the lung lesions, dyspnea, thoracalgia, mediastinal lymphadenopathy, and pleural effusion did not significantly predict overall survival. There was no significant difference in prognosis according to the type of treatment.@*CONCLUSIONS@#T. marneffei infections involving the respiratory system are common. The critical determinants of prognosis are HIV infection, CD4/CD8, percentage of CD4 T cells, type of treatment, and the time range from onset to confirmation of diagnosis. Rapid and accurate diagnosis is crucial for improving prognosis.

Determinants of prognosis in Talaromyces marneffei infections with respiratory system lesions / 中华医学杂志(英文版)

Background@#Little study has investigated the differences between Talatomyces marneffei (T. marneffei) respiratory infection and tuberculosis and the prognostic factors of such infection. This study investigated the characteristics and prognostic factors of T. marneffei infections with respiratory lesions and the causes of misdiagnosis.@*Methods@#Clinical characteristics and prognoses of patients with T. marneffei infections with respiratory system lesion were investigated. T. marneffei diagnosis followed isolation from clinical specimens using standard culture, cytology, and histopathology. Survival curves were estimated by using Kaplan-Meier analysis, with log-rank test to compare differences in survival rates between groups. Univariate and multivariate Cox regression analyses were also performed to assess significant differences in clinical characteristics of overall survival.@*Results@#Of 126 patients diagnosed with T. marneffei infections, 63 (50.0%) had T. marneffei respiratory system infections; 38.1% (24/63) were misdiagnosed as having tuberculosis. Human immunodeficiency virus (HIV) infection, CD4/CD8 < 0.5, percentage of CD4+ T cells <42.8%, and length of time from onset to confirmation of diagnosis >105 days were potential risk factors for poor prognoses. Length of time from onset to confirmation of diagnosis persisted as an independent predictor of all-cause mortality in multivariate analysis (odds ratio: 0.083, 95.0% confidence interval: 0.021–0.326, P < 0.001). However, the size of the lung lesions, dyspnea, thoracalgia, mediastinal lymphadenopathy, and pleural effusion did not significantly predict overall survival. There was no significant difference in prognosis according to the type of treatment.@*Conclusions@#T. marneffei infections involving the respiratory system are common. The critical determinants of prognosis are HIV infection, CD4/CD8, percentage of CD4+ T cells, type of treatment, and the time range from onset to confirmation of diagnosis. Rapid and accurate diagnosis is crucial for improving prognosis.

Tissue Distribution of 14 Bioactive Components After Oral Administration of Gegen Qinlian Tang in Rats / 中国实验方剂学杂志

Objective:To investigate the tissue distribution of major bioactive components from Gegen qinlian Tang(GQT) in rats,and to reveal the mechanism for the efficacy of GQT by the tissue targeting of its bioactive ingredients in vivo. Method:After oral administration of GQT in rats,tissues were collected at different time points,including small and large intestine,liver,heart,spleen,lung,and kidney.Samples were prepared for determination of 14 bioactive components of GQT in tissue homogenate by HPLC-MS/MS.The chromatography separation was performed on an Agilent ZORBAX SB-C18 column(2.1 mm×100 mm,3.5 μm) with acetonitrile-0.1% formic acid in water for gradient elution.Electrospray ionization(ESI) was applied and operated in the positive ion mode.Meanwhile,naringin was used as the internal standard for determining. Result:HPLC-MS/MS for simultaneous determination of 14 components from GQT in tissue homogenate was developed and validated by specificity,calibration curves,recovery test,matrix effect,precision,accuracy,and stability.In the small intestine,the the area under the curve(AUC0-10 h) of major isoflavonoids(puerarin,3'-hydroxypuerarin,and daidzein) were 22 174.9,15 893.1,3 882.5 h·mg·L-1,major flavonoids(baicalein,wogonin,wogonoside,and baicalin) were 15 423.6,15 408.4,7 017.3,3 697.7 h·mg·L-1,and major alkaloid(berberine) was 2 544.0 h·mg·L-1,respectively.The distribution of these ingredients in the small intestine was significantly higher than that in other tissues. Conclusion:The effective ingredients of GQT are mainly distributed in the intestinal tissues,which speculates that the anti-inflammatory and antidiarrheal activities of GQT may be related to its targeting in the intestine.

Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics / 中国实验方剂学杂志

Objective: To study on the antitumor mechanism of artesunate in the treatment of liver cancer based on gas chromatography-mass spectrometry (GC-MS). Method: CellTiter-Glo® Luminescent Cell Viability Assay was used to detect activity of artesunate with different concentrations (0, 12.5, 25, 50, 100 μmol·L-1) on human liver cancer Huh7, SMMC-7721 cells for 24, 48, 72 h. GC-MS was employed to analyze the changes of metabolites of artesunate in two kinds of hepatoma cells (Huh7, SMMC-7721) for 24 h. The data was preprocessed by Postrun Analysis 4.41 workstation. Partial least squares-discriminant analysis (PLS-DA) was used to analyze two sets of differential metabolites and to analyze metabolic pathways of differential metabolites based on MetaboAnalyst 3.0 software. Result: Compared with the normal group, after two kinds of liver cancer cells was treated by artesunate, a total of 39 identical metabolites in the cells have undergone significant changes, which were mainly related to five metabolic pathways,including biosynthesis of aminoacyl-transfer RNA (tRNA), metabolism of alanine, aspartic acid and glutamic acid, metabolism of glycine, serine and threonine, metabolism of arginine and proline, metabolism of glutathione. Conclusion: Artesunate (12.5-100 μmol·L-1) can inhibit the growth of liver cancer cells (Huh7, SMMC-7721), it mainly involves five metabolic pathways, which may be the pathway of artesunate against liver cancer.

Changes in body mass index of postural tachycardia syndrome in children and adolescents / 中国实用儿科杂志

OBJECTIVE: To discuss the relationship between body mass index and postural tachycardia syndrome in children and adolescents. METHODS: The clinical data of 127 children and adolescents were retrospectively analyzed,who were diagnosed with postural tachycardia syndrome(POTS)in the Department of Pediatric Cardiovasology,Children's Medical Center,the Second Xiangya Hospital,Central South University,from August 2009 to June 2018,which included63 males and 64 females and their ages were from 4 to 17 years old(mean age 11.31±2.53)(POTS group). A total of 107 healthy children and adolescents including 64 males and 63 females were included as the control group,who had a health examination in the health care clinic in the hospital during the same period(aged 7 to 17,with a mean age of11.60±3.27). Body length and body mass were measured and body mass index(BMI)calculated. Statistical analysis was conducted with SPSS 22.0 software. RESULTS:(1)Duration and frequency of syncope:the duration of syncope was(8.13±13.76)months in POTS group and the frequency of syncope was(1.45±4.43)times.(2)BMI intergroup comparison:comparing POTS group with control group,there was no difference in age,length or body mass(P>0.05),and BMI was significantly lower[(17.32 ± 2.65)kg/m~2 vs.(18.17 ± 2.42)kg/m~2,t=2.655,P<0.01]in POTS group.(3)BMI classification:low body mass was higher in POTS group(69.29%,88/127)than in control group(56.69%,72/127);normal body mass was lower in POTS group(29.13%,37/127)than in control group(41.73%,53/127）,χ~2=4.444,P<0.05. CONCLUSION: BMI of POTS group decreases significantly in children and adolescents,and it is lower in girls than in boys.

Expression and Clinical Significance of STAT3 Genes in Patients with Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression of STAT3 gene in patients with acute myeloid leukemia and its correlation with clinical characteristics.@*METHODS@#The real-time quantitative RT-PCR was used to detect the level of STAT3 mRNA in bone marrow samples from 38 newly diagnosed patients with acute myeloid leukemia(AML), and its relevance with clinical characteristics and prognosis were statistically analyzed. Western blot was employed to detect the STAT3 protein level in AML patients. The bone marrow cells from 15 healthy subjects were used as control.@*RESULTS@#At the mRNA level, the expression level of STAT3 in the AML group was significantly higher than that in control group (P0.05). The median survival time of patients in STAT3 low expression group was logner than that in high expression group, but the difference was not statistically significant (P>0.005). The level of STAT3 protein in AML patients was significantly higher than that in control group (P<0.05）.@*CONCLUSION@#The STAT3 gene is highly expressed in AML patients, which may be used as a predictor for high-risk of AML.

Effect of Silencing LNK Gene on the Expression of EPO and EPOR in THP-1 Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of silencing LNK gene on the expression of EPO and EPOR in acute myeloid leukemia cells (THP-1).@*METHODS@#THP-1 cells were cultured. The lentivirus was used as a vector to silence the LNK gene stably. After 72 hours of infection, GFP expression level was detected by the fluorescent inverted microscopy. The lentiviral Infection efficiencies were monitored by flow cytometry. The LNK silencing effect was confirmed. The mRNA expressions of EPO and EPOR were detected by RT-PCR. The protein levels of LNK, EPO and EPOR were detected by Western blot.@*RESULTS@#At the time-point of 72 hours after lentivirus infection, the expression level of GFP was above 85% detected by fluorescent inverted microscopy. The infection efficiency was above 99% by flow cytometry. mRNA expressions of LNK, EPO and EPOR in LNK silencing group were signifycantly lower than those in control group (P＜0.05). The protein levels of LNK, EPO and EPOR in LNK silencing group were significantly lower than those in the control group (P＜0.05).@*CONCLUSION@#THP-1 cell line of LNK gene silencing has been successfully established,the LNK gene has been silenced, the expression of EPO and EPOR decrease, indicating that LNK may participate in the regulation of EPO and EPOR.

Pharmacokinetic characteristics of fourteen main components of Gegen Qinlian Decoction in rats / 中国中药杂志

A specific and selective UPLC-MS/MS method was developed and validated for the simultaneous determination of isoflavonoids(3'-hydroxy puerarin, puerarin, daidzin, daidzein, genistin, genistein), flavonoids (baicalin, baicalein, wogonoside, wogonin, liquiritin)and alkaloids(berberine, jatrorrhizine, palmatine)(14 bioactive compounds) of Gegen Qinlian Decoction(GQD) in plasma. The pharmacokinetics characteristics of 14 bioactive compounds were study after oral administration of GQD at a single dose to rats. Prednisolone was used as the internal standard of liquiritin, and naringin was used as the internal standard of the other thirteen analytes. After the plasma samples were processed by precipitation protein method, the constituents and internal standards were gradient eluted by using a Zorbax SB-18 column with a mobile phase of acetonitrile(A) and 0.1% formic acid aqueous solution(B) using a gradient elution of 0-2.5 min, 15%-30% A; 2.5-3.5 min, 30%-35% A; 3.5-5.0 min, 35%-40% A; 5.0-9.0 min, 40%-60% A; 9.0-11.0 min, 60%-15% A, and the flow rate was 0.4 mL·min⁻¹. The auto sampler was conditioned at 25 °C and the sample injection volume was 5 μL. A mass spectrometry was applied with electrospray ionization (ESI) ion source in the positive and negative ion multiple reaction monitoring(MRM) mode. All pharmacokinetic parameters were processed by non-compartmental analysis with DAS 3.2.2 software. The results showed that the linear correlation coefficient of the 14 components were all greater than 0.99, indicating that the method had good linearity in their respective concentration ranges. Post-preparative stability (25 °C, 24 h), short-term stability(25 °C, 12 h), long-term stability (-20 °C, 7 d), and freeze and thaw stability (3-cycles) of the fourteen constituents were examined to evaluate the stability of methodology. The results of the inner and inter-day relative standard deviations were both less than 10%, indicating legitimate precise and accuracy to the requirement of biological sample analysis. The assay method is proved to be sensitive, accurate and convenient. It can be applied to the pharmacokinetic study of the fourteen analytes. The kinetic parameters of the related drugs were calculated according to the blood concentration of the 14 components. The results showed that the MRT0-t of the isoflavones and flavonoids was 7.5-11.8 h, T1/2z were mainly in 11.0-29.7 h, and the AUC0-t flavonoids were larger than the isoflavones. The MRT0-t of alkaloids were between 4.3-7.2 h, T1/2z were 1.0-5.0 h, AUC0-t were less than flavonoids and isoflavones. The results suggest that flavonoids and isoflavones have a high concentration of blood and long time of action, which are beneficial to the anti-inflammatory and antipyretic effects. The concentration of alkaloids in the body is low and the time of action is short, and it may play its bacteriostasis in the intestinal tract.

Correlation of tyrosine phosphatase-1/2 (SHP-1,SHP-2) with indoleamine 2,3-dioxygenase (IDO) in villi and decidua of early pregnancy / 中国免疫学杂志

Objective:To explore the correlation of tyrosine phosphatase-1/2 (SHP-1,SHP-2) with indoleamine 2,3-dioxygenase(IDO) in maternal fetal interface.Methods: The expression of SHP-1,SHP-2 and IDO were detected by Western blot method and the relationship of the proteins was analysed,in human chorionic villi and decidua tissues of 30 cases of artificial abortion patients.Results:The expression of SHP-1,SHP-2 were positively correlated withthe expression of IDO in human chorionic villi and de-cidua;the expression of SHP-1,SHP-2 and IDO in decidual tissues were higher than those in the villi.Conclusion: Normal physiological state of pregnancy,SHP-1 and SHP-2 may be involved in the regulation of immune tolerance by positive regulation of IDO expression at maternal fetal interface.

Expression of LNK Genes in Patients with Acute Leukemia and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of LNK gene in patients with acute leukemia (AL) and its correlation with the clinical characteristics.</p><p><b>METHODS</b>Real-time quantitative RT-PCR was used to detect the level of LNK mRNA in bone marrow samples from 80 patients diagnosed as AL(42 cases of ALL, and 38 cases of AML), and its relevance with clinical indicators was statistically analyzed. Western blot was used to detect the expression of LNK protein. The bone marrow samples of 16 healthy volunteers were used as the controls.</p><p><b>RESULTS</b>The LNK mRNA levels in ALL and AML groups were significantly higher than that in control group (P=0.007, P=0.021) and there was no statistical difference between ALL and AML groups. The LNK levels in ALL and AML groups possitively correlated with the risk of patients (P=0.000, P=0.04, r=0.5, r=0.386), And the LNK levels in high risk ALL and AML groups were significantly higher than that in control group (P=0.035, P=0.032), the LNK levels in intermediate risk of AML and ALL groups (P=0.239,P=0.609) and the LNK level in standard risk (P=0.974, P=1) were all higher than that in control group, there was no statistianl significance. but the risks of different groups showed no statistical significance. The LNK protein level in patients with acute leukemia was higher than that in control group.</p><p><b>CONCLUSION</b>The expression level of LNK gene in AL patients is higher than that in healthy people, and the expression level of LNK gene positively correlates with the risk of patients.</p>

Utility of Droplet Digital PCR Assay for Quantitative Detection of Norovirus in Shellfish, from Production to Consumption in Guangxi, China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Shellfish are recognized as important vehicles of norovirus-associated gastroenteritis. The present study aimed to monitor norovirus contamination in oysters along the farm-to-fork continuum in Guangxi, a major oyster production area in Southwestern China.</p><p><b>METHODS</b>Oyster samples were collected monthly from farms, markets, and restaurants, from January to December 2016. Norovirus was detected and quantified by one-step reverse transcription-droplet digital polymerase chain reaction (RT-ddPCR).</p><p><b>RESULTS</b>A total of 480 oyster samples were collected and tested for norovirus genogroups I and II. Norovirus was detected in 20.7% of samples, with genogroup II predominating. No significant difference was observed in norovirus prevalence among different sampling sites. The norovirus levels varied widely, with a geometric mean of 19,300 copies/g in digestive glands. Both norovirus prevalence and viral loads showed obvious seasonality, with a strong winter bias.</p><p><b>CONCLUSION</b>This study provides a systematic analysis of norovirus contamination 'from the farm to the fork' in Guangxi. RT-ddPCR can be a useful tool for detection and quantification of low amounts of norovirus in the presence of inhibitors found particularly in foodstuffs. This approach will contribute to the development of strategies for controlling and reducing the risk of human illness resulting from shellfish consumption.</p>

Variation of STAT3 Gene in Myleproliferative Neoplasms and Its Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To detect the mutation and single nucleotide polymorphisms of STAT3 gene in the patients with myeloproliferative neoplasms (MPN), and to analyze the correlation between STAT3 gene and the subtypes of MPN.</p><p><b>METHODS</b>A total of 147 patients with MPN were selected, including 28 patients with polycythaemia vera (PV), 46 patients with essential thrombocythemia (ET), 10 patients with primary myelofibrosis (PMF), and 63 patients with chronic myeloid leukemia (CML); and 88 healthy persons were used as normal control. DNA of all cases was extracted from bone marrow or peripheral blood, and JAK2V617F gene mutation was detected by allele-specific PCR, then 23 exons of STAT3 gene were amplified by PCR. Mutation and single nucleotide polymorphism of Rs2293152 of STAT3 gene were identified by DNA sequencing.</p><p><b>RESULTS</b>STAT3 gene mutation was found in 8 patients with CML. The mutation rate was 12.7%. the missense mutation（S629T）as found in 3 cases, the synonymous mutation was found in 5 cases (Q469Q 3 cases, G618G 2 cases). One case had mutations at the both sites of S629T and G618G. No mutation of STAT3 gene was found in the normal control group. Rs2293152: detection showed that the G allele of CML group was significantly higher than that of normal control, PV, ET and PMF group (P<0.01), suggesting that the patients with Rs2293152 G allele were more likely to develop CML. The C allele of PV, ET and PMF group was significantly higher than that of CML group (P<0.05), suggesting that the patients with Rs2293152 C allele were more likely to develop PV, ET and PMF. The G allele fiequency of JAK2V617F-negative group was significantly lower than that of the normal control and JAK2V617F positive group (P<0.01), suggesting that the Rs2293152 G allele may be a factor protecting against JAK2V617F mutation.</p><p><b>CONCLUSION</b>In MPN patients, STAT3 gene is unstable and prone to mutation. The different alleles of the Rs2293152 locus of the STAT3 gene are relates with different subtypes and JAK2V617F-negative MPN.</p>

Artesunate Suppresses Cell Proliferation of THP-1 Cells by Inducing Apoptosis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate whether Artesunate(ART) can inhibit the proliferation of THP-1 cells and to explore the potential mechanism of its anti-leukemia effect.</p><p><b>METHODS</b>THP-1 cells were treated with 5 concentrations of Artesunate for 24 h, 48 h or 72 h. The viability of cells was detected with CCK-8 assay, apoptosis was assessed by using flow cytometry, and the STAT3, Caspase3 and Caspase8 protein levels were measured with Western blot .</p><p><b>RESULTS</b>Compared with the control group, ART significantly inhibited the proliferation of THP-1 cells in a dose-dependent manner (r=0.9829, P<0.05). ART also increased the apoptosis of THP-1 cells. The results of Western blot showed that after treated with ART, the STAT3 protein expression in THP-1 cells was significantly down-regulated (P<0.01), and the expressions of Caspase3, cleaved Caspase3 and Caspase8 proteins were up-regulated(P<0.01).</p><p><b>CONCLUSION</b>Artesunate can inhibit the proliferation of THP-1 cells, which may relate with the down-regulation of STAT3 expression and the activation of Capase3 and Caspase8.</p>