Content determination of ten flavonoids and alkaloids in Gleditsiae Sinensis Fructus, Gleditsiae Fructus Abnormalis, and Gleditsiae Spina / 中国中药杂志

To study the quality control of three traditional Chinese medicines derived from Gleditsia sinensis [Gleditsiae Sinensis Fructus(GSF), Gleditsiae Fructus Abnormalis(GFA), and Gleditsiae Spina(GS)], this paper established a multiple reaction monitoring(MRM) approach based on ultra-high performance liquid chromatography-triple quadrupole-linear ion-trap mass spectrometry(UHPLC-Q-Trap-MS). Using an ACQUITY UPLC BEH C_(18) column(2.1 mm × 100 mm, 1.7 μm), gradient elution was performed at 40 ℃ with water containing 0.1% formic acid-acetonitrile as the mobile phase running at 0.3 mL·min~(-1), and the separation and content determination of ten chemical constituents(e.g., saikachinoside A, locustoside A, orientin, taxifolin, vitexin, isoquercitrin, luteolin, quercitrin, quercetin, and apigenin) in GSF, GFA, and GS were enabled within 31 min. The established method could quickly and efficiently determine the content of ten chemical constituents in GSF, GFA, and GS. All constituents showed good linearity(r>0.995), and the average recovery rate was 94.09%-110.9%. The results showed that, the content of two alkaloids in GSF(2.03-834.75 μg·g~(-1)) was higher than that in GFA(0.03-10.41 μg·g~(-1)) and GS(0.04-13.66 μg·g~(-1)), while the content of eight flavonoids in GS(0.54-2.38 mg·g~(-1)) was higher than that in GSF(0.08-0.29 mg·g~(-1)) and GFA(0.15-0.32 mg·g~(-1)). These results provide references for the quality control of G. sinensis-derived TCMs.

Standardized Jin's three-needle therapy for stroke: a randomized controlled trial / 中国针灸

OBJECTIVE@#To observe the effect of standardized Jin's three-needle therapy on limb motor function and nerve function defect in stroke patients, and to evaluate the placebo control method.@*METHODS@#A total of 66 patients with stroke were randomly divided into a Jin's three-needle group (33 cases, 3 cases dropped off) and a placebo needle group (33 cases, 4 cases dropped off). All the patients were treated with conventional medication and rehabilitation treatment. In addition, the patients in the Jin's three-needle group were treated with standardized Jin's three-needle therapy at temporal three points, spirit four points, hand three points, foot three points, upper extremity spasm three points, lower extremity spasm three points, etc.; while the patients in the placebo needle group were treated with placebo needling at identical points. All the treatments were given once a day, 5 days a week, and 3-week treatment was given with an interval of 2 days between weeks. The scores of Fugl-Meyer assessment scale (FMA) and National Institutes of Health stroke scale (NIHSS) were observed before treatment, 10 d and 21 d into treatment, and the blind evaluation was conducted after treatment.@*RESULTS@#On the 10 d and 21 d into treatment, the FMA scores in both groups were higher than those before treatment (P<0.01), and the NIHSS scores were lower than those before treatment (P<0.01). On the 10 d and 21 d into treatment, the FMA scores in the Jin's three-needle group were higher than those in the placebo needle group (P<0.05); on the 10 d into treatment, the NIHSS score in the Jin's three-needle group was were lower than that in the placebo needle group (P<0.05). There was no significant difference between the two groups on judging the type of treatment (P>0.05), and the consistency with the real situation was poor (Cohen's kappa coefficient<0.20).@*CONCLUSION@#The standardized Jin's three-needle therapy could effectively improve the limb motor function and nerve function defect in stroke patients. The placebo control method used in this study shows good clinical operability and masking effect.

LASS2/TMSG1 overexpression inhibits proliferation and promotes apoptosis of human lung cancer A549 cells possibly by upregulating ceramide and p38 MAPK to activate a signaling cascade / 南方医科大学学报

OBJECTIVE@#To investigate the effects of LASS2/TMSG1 gene overexpression on proliferation and apoptosis of human lung cancer A549 cells and explore the possible mechanism.@*METHODS@#We examined LASS2/TMSG1 expression level in a previously constructed A549 cell line overexpressing LASS2/TMSG1 using Western blotting. The proliferation and apoptosis of the cells were detected using colony-forming assay, CCK-8 assay, Hoechst/PI double staining and flow cytometry. Fourteen nude mice were randomized into 2 groups (n=7) to receive subcutaneous injection of A549 cells with or without LASS2/TMSG1 overexpression on the back of the neck, and the cell proliferation in vivo was observed. The expression levels of p38 MAPK protein and p-p38 MAPK protein in the xenografts were detected with Western blotting. ELISA was used to detect the levels of ceramide and p38 MAPK protein in cultured A549 cell supernatants and the xenografts in nude mice.@*RESULTS@#Compared with the negative control cells, A549 cells with LASS2/TMSG1 overexpression had significantly lowered proliferation ability in vitro with increased early apoptosis rate (P < 0.05), and showed obvious growth inhibition after inoculation in nude mice(P < 0.05). Western blotting showed that in both cultured A549 cells and the xenografts in nude mice, LASS2/TMSG1 gene overexpression significantly increased the expression levels of p38 MAPK protein and p-p38 MAPK protein (P < 0.05); the results of ELISA also revealed significantly increased levels of ceramide and p38 MAPK protein in the cell supernatant andxenografts as well (P < 0.05).@*CONCLUSION@#Overexpression of LASS2/TMSG1 gene can significantly inhibit the proliferation and promote early apoptosis of human lung cancer A549 cells both in vitro and in vivo possibly by upregulating the expressions of ceramide and p38 MAPK protein to activate a signal transduction cascade.

A prospective randomized controlled study on mouse nerve growth factor in the treatment of global developmental delay in children / 中国当代儿科杂志

OBJECTIVES@#To study the clinical effect of mouse nerve growth factor (mNGF) in the treatment of children with global developmental delay (GDD).@*METHODS@#A prospective clinical trial was conducted in 60 children with GDD who were treated in the First Affiliated Hospital of Anhui Medical University between July 2016 and July 2017. These children were randomly divided into two groups: conventional rehabilitation treatment and mNGF treatment group (@*RESULTS@#Before treatment and after 1.5 months of treatment, there was no significant difference in the developmental quotient (DQ) of each functional area of the Gesell Developmental Scale between the mNGF treatment and conventional rehabilitation treatment groups (@*CONCLUSIONS@#In children with GDD, routine rehabilitation training combined with mNGF therapy can significantly improve their cognitive, motor, and social abilities.

Ganoderma lucidum polysaccharides promote T lymphocyte infiltration into tumor by regulating expression of ICAM-1 in endothelial cells / 中国中药杂志

Polysaccharide is among the main active components of Ganoderma lucidum for tumor prevention and treatment. Howe-ver, it remains unclear whether it has synergy with tumor immunotherapy. This study evaluated the effect of G. lucidum polysaccharides(GLP) on the infiltration of T lymphocytes into tumor and the underlying mechanism, in order to provide a reference for its application in tumor immunotherapy. GLP were prepared by water extraction and alcohol precipitation combined with Sevag method and then given(intraperitoneal injection) to the mice bearing B16-F10 cells at 25, 50 and 100 mg kg~(-1), respectively, to evaluate the effect on tumor growth. The infiltration of CD3~+ and CD8~+ T cells and the expression of intercellular cell adhesion molecule-1(ICAM-1) in tumor were detected by immunohistochemistry. EA.hy926 cells were treated with 50, 100 and 200 μg·mL~(-1) GLP, and the expression of ICAM-1 was determined by Western blot. The adhesion of EA.hy926 cells treated with GLP was measured with fluorescence-labeled Jurkat cells. To analyze the mechanism based on NF-κB pathway, this study determined the protein levels of nuclear factor kappa-B(NF-κB) p65, alpha inhibitor of NF-κB(IκBα), p-NF-κB p65 and p-IκBα by Western blot. The results showed that GLP can significantly inhibit the tumor growth in mice bearing B16-F10 cells, promote the infiltration of CD3~+ and CD8~+ T cells in tumor, and increase the expression of ICAM-1 in tumor. Meanwhile, GLP could also enhance the expression of ICAM-1 in EA.hy926 cells, thus strengthen the adhesion to Jurkat cells, induce phosphorylation and protein degradation of IκBα, and raise the expression and phosphorylation level of NF-κB p65. These results suggested that GLP could promote the expression of ICAM-1 through NF-κB pathway and further enhance the infiltration of T lymphocytes into tumor, thereby inhibiting tumor growth. This study lays a foundation for the further application of GLP in tumor immunotherapy.

Progress in immunomodulatory effects and their mechanisms of insect antimicrobial peptides / 医学研究生学报

Insect antimicrobial peptides (AMPs) are small cationic molecules with various biological activities induced by many factors. Insect AMPs, which are essential components of insect innate immune system, may have functionsof anti-infection, anti-tumor and immune modulation in the medicine field. Many different types of insect AMPs, such as cecropin-like AMPs extracted from Saturniidae, defensin-like AMPs purified from Catharsius molossus L., and Pro-rich and Gly-rich peptides from drosophilid fly and silkworm, respectively, have been identified, and been reported to have anti-infective and anti-tumor functions. However, relatively few reports have focused on the immunomodulatory activities and their possible mechanisms of insect AMPs. It has been found that insect AMPs could activate immune cells, enhance the activities of NK cells and cytotoxic T lymphocytes, induce cytokine production, and inhibit the activities of inflammatory cytokines and inflammatory mediators. Furthermore, Toll-like receptor pathways, nuclear factor-κB signaling pathway and mitogen-activated protein kinase signaling pathway were also reported to be involved in their immunomodulatory mechanisms. In this paper, the immunomodulatory effects of some classes of insect AMPs (Lepidoptera, Diptera, Coleoptera, and Hymenoptera are involved),and their signaling pathway - related mechanisms are reviewed. It was hoped that these data could provide references for the future research in the field of insect AMPs.

Clinical phenotypes and genetic features of families with Duchenne muscular dystrophy / 中国当代儿科杂志

OBJECTIVE@#To study the phenotypes and genetic features of families with Duchenne muscular dystrophy (DMD).@*METHODS@#Seven children from six families with DMD diagnosed by gene testing were enrolled. The clinical and genetic features of the families were analyzed.@*RESULTS@#There were two new mutations and four maternal inheritance mutations in the six families. The proband of family 1 had one point de novo mutation and one insertion de novo mutation of the DMD gene. Three families had point mutation, one family had fragment deletion of exon, and one family had fragment duplication of exon. The youngest age of onset of the probands was 6 months. All probands had skeletal muscle dyskinesia and significant changes in muscle enzymes, with different severities of clinical phenotypes. Three probands had mild mental retardation. The results of echocardiography were normal for all probands. The mother of the proband in family 6 had mild clinical phenotype.@*CONCLUSIONS@#Gene testing can be used for the confirmed diagnosis of DMD. Mental retardation is a frequent clinical phenotype of DMD. The symptoms of myocardial involvement are not obvious in the early stage. Female carriers may have mild clinical symptoms.

MiR-30a-5p Inhibits Epithelial Mesenchymal Transition Function in Human Cervical Cancer Cells by Targeting Ubiquitin-specific Protease 22 / 中山大学学报(医学科学版)

@# 【Objective】To investigate the inhibitory effect and mechanism of microRNA-30a-5p（miR-30a-5p）on epithelial mesenchymal transition in cervical cancer Hela cells.【Methods】Hela cervical cancer cell lines were transfected with miR-30a-5p mimics or negative control mimic，respectively，as 30a-5p or NC group. Control group was established with untreated Hela cervical cancer cells. miR-30a-5p content in each group was detected by RT-PCR assay. Transwell assay was used to detect the invasion ability of the 3 groups. Western-blot assay was used to detect the expressions of N- cadherin，α-Catenin and ubiquitin specific processing peptidase 22（USP22）protein in the 3 groups. Prediction target genes of miR-30a-5p by bioinformatics methods. Antagonistic effect of USP22 over-expression on miR-30a-5p inhibi⁃tion of EMT was detected by western blot assay. The relationship between miR-30a-5p and USP22 was detected by dual luciferase assay. Subcutaneous transplantation tumor model established，and the effect of miR-30a-5p in vivo was ob⁃ served.【Results】The miR-30a-5p intracellular quantity in 30a-5p group Hela cells was up-regulated，and the expres⁃ sion level of miR-30a-5p was 853.82（862.26~843.11）times higher than that of Control group（P＜0.01）. The number of invasive cells in 30a-5p group was 8.17（8.32~8.03），which was significantly lower than that of Control group 62.33 （63.52~60.19）（P＜0.01）. USP22 may be the target gene of miR-30a-5p. In 30a-5p group，the intracellular quantity of N-cadherin protein was decreased，the intracellular quantity of α-Catenin protein was increased，and the intracellular quantity of USP22 protein was decreased. The intracellular quantity of N-cadherin protein in 30a-5p group was down-reg⁃ ulated，the intracellular quantity of α-Catenin protein was increased，and the intracellular quantity of USP22 protein was reduced. After USP22 over-expression，the intracellular quantity of N-cadherin protein in cervical cancer cells of 30a-5p group was up-regulated，and the intracellular quantity of α-Catenin protein were down-regulated. Dual luciferase assay showed that USP22 is a downstream target gene of miR-30a-5p（P＜0.01）. Subcutaneous transplantation tumors in 30a- 5p group were significantly smaller than those in Control group.【Conclusion】miR-30a-5p may inhibit the expression of EMT related protein through the downstream target gene USP22，and the epithelial mesenchymal transition function of cervical cancer Hela cells.

The Clinical Analysis of Necrotizing Enterocolitis / 昆明医科大学学报

Objective From May 2010 to May 2013 and from June 2013 to June 2016, we compared the clinical analysis of patients with necrotizing enterocolitis (NEC) in the Department of Pediatrics, studied incidence rate of NEC and the relationship between stages of NEC and prognosis, and compared the prognosis between the two three-year periods to provide the experience of diagnosis and treatment for clinical practice in the future. Methods The clinical data of patients diagnosed with NEC were retrospectively analyzed according to the diagnostic criteria of modified Bell staging classification using SPSS statistical software package. Early diagnosis rate and prognosis were compared between the two three-year periods. P＜0.05 showed that statistical significance was found. Results From May 2010 to May 2013, 40 cases of NEC were diagnosed in our hospital, among which 8 was in the first stage (20%),24 in the second stage (60%),and 8 in the third stage (20%) . The number of premature infants was 32, accounting for 80% , among which 25 cases were infants with very low birth weight accounting for 62.5% and 8 cases were full-term infants, accounting for 20% (P＜0.05) . In the study, 32 cases (80%) received traditional treatment, which was effective in stage Ⅰ. During the second stage, two abandoned the treatment was confirmed death in the follow-up, two underwent surgery successfully due to the progression of the disease and four (25%) died in the hospital. Statistic significance of treatment of internal medicine was found when comparing stage Ⅰ with stage Ⅱ (P＜0.05) . Ten children underwent surgery, among which one gave up treatment after the surgery because of financial problems and was found dead in the follow-up and four (60%) died in the hospital. The comparison of surgical treatment in patients in stage Ⅱ and Ⅲ showed no statistical significance (P＞0.05) . Fatality rates were statistically significant (P＜0.05) . From June 2013 to June 2016, 41 children were diagnosed with NEC in our hospital, including 24 cases in stage Ⅰ (59%), 14 in stage Ⅱ (34%), and 3 in stage Ⅲ (7%) . The number of premature infants was 33, accounting for 80%, among which 26 cases were infants with very low birth weight accounting for 62.5% and 8 cases were full-term infants, accounting for 20% (P＜0.05) . In the study, 35 cases (85.3%) received traditional treatment, which was effective in stage Ⅰ. During the second stage, one underwent surgery successfully due to the progression of the disease. Statistic significance of treatment of internal medicine was found when comparing stage Ⅰ with stage Ⅱ (P＜0.05) . Four children underwent surgery, among which one gave up treatment after the surgery because of financial problems and was found dead in the follow-up and the surgery was successful in other three. The comparison of surgical treatment in patients in stage Ⅱ and Ⅲ showed statistical significance (P＜0.05) . Fatality rates were statistically significant comparing the two three-year periods (P＜0.05) . The comparison of the rates of early diagnosis in the two three-year periods showed statistically significance (P＜0.001) . Conclusion Early diagnosis of NEC is the key to reduce mortality. Intestinal fatty acid binding protein (I-FABP) is a serological marker for early diagnosis of NEC. As an important factor, the reduction of the incidence of premature birth will result in the drop in the incidence of NEC.

Genome-Wide Screening of Aberrant Methylation Loci for Nonsyndromic Cleft Lip / 中华医学杂志(英文版)

Background@#The pathogenicity of cleft lip (CL) is pretty complicated since it is influenced by the interaction of environment and genetic factors. The purpose of this study was to conduct a genome-wide screening of aberrant methylation loci in partial lesion tissues of patients with nonsyndromic CL (NSCL) and preliminarily validate candidate dysmethylated genes associated with NSCL.@*Methods@#Fifteen healthy and sixteen NSCL fetal lip tissue samples were collected. The Infinium HumanMethylation450 BeadChip was used to screen aberrant methylation loci in three NSCL and three healthy lip tissues. The differential methylation sites and functions of the annotated genes between NSCL and healthy lip tissues were analyzed using minfi package of R software, cluster analysis, Gene Ontology (GO) annotation, and metabolic pathway annotation. Gene expression was assessed in nine differentially methylated genes by real-time polymerase chain reaction (PCR). The transcriptions mRNA levels of three out of nine candidate genes were downregulated remarkably in NSCL lip tissues, and these three genes' abnormal methylation loci were validated by pyrosequencing in 16 NSCL cases and 15 healthy cases.@*Results@#In total, 4879 sites in the genes of NSCL odinopoeia fetuses showed aberrant methylation when compared with normal lip tissue genome. Among these, 3661 sites were hypermethylated and 1218 sites were hypomethylated as compared to methylation levels in healthy specimens. These aberrant methylation sites involved 2849 genes and were widely distributed among the chromosomes. Most differentially methylated sites were located in cytosine-phosphoric acid-guanine islands. Based on GO analysis, aberrantly methylated genes were involved in 11 cellular components, 13 molecular functions, and a variety of biological processes. Notably, the transcription of DAB1, REELIN, and FYN was significantly downregulated in lesion tissues of NSCL fetus (P < 0.05). Pyrosequencing results validated that there were two loci in DAB1 with high methylation status in patient tissues (P < 0.05).@*Conclusions@#We detected numerous aberrantly methylated loci in lesion tissues of NSCL fetus. Aberrant gene expression in the REELIN signaling pathway might be related with NSCL. Decreased transcription of DAB1, a member of REELIN signal pathway, resulted from its abnormal high methylation, which might be one of the factors underlying the occurrence of NSCL.

Analysis of the clinical treatment and the influence of compliance on amblyopia / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To explore the treatment methods and curative effect of amblyopia in children, and to analyze the effect of compliance on the treatment process.METHODS:The children with amblyopia treatment since January 1st 2010 in General Hospital of Jinan Military Area, were selected and completed follow-up for a total of 129 eyes of 78 patients, including 53 eyes of male in 31 cases and 76 eyes of female in 47 cases, the initial age of 3 to 12 years, the treatment were performed with accurate optometry, reasonable glasses and appropriate to cover the dominant eye based comprehensive therapy and compliance training of the parents and children.The treatment time was 12-36mo, followed up for 3a.The final therapeutic effect was observed and the difference of the stage curative effect between the children with different grade of compliance (good, moderate and poor grade) was compared.RESULTS:In the treatment of 36mo, 106 eyes were cured, 21 eyes were essentially recovered and the total cure rate was 98.4%, which was improved in 2 eyes (1.6%) and was ineffective in 0 eyes (0%);the total effective rate were 100%.The total cure rate was 69% in the treatment of 6mo and there was a significant difference in the cure rate among children with different compliance (overall comparison P<0.001, each comparison P<0.017).In the treatment of 12mo, the total cure rate was 89.1% and the cure rate were significantly different between children with the moderate compliance and the children with good compliance (P<0.05).CONCLUSION:Comprehensive treatment of children with amblyopia can achieve an ideal clinical efficacy.Compliance is an important factor influencing the treatment effect and the treatment process.The parents and children`s compliance training can shorten the course of treatment and improve the curative effect.

NKX2.5 and TBX5 gene mutations in in vitro fertilization children with congenital heart disease / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the differences of NKX2.5 and TBX5 gene mutations between in vitro fertilization (IVF) children with congenital heart disease (CHD) and naturally conceived children with CHD.</p><p><b>METHODS</b>Blood samples from 68 IVF children with CHD and 98 naturally conceived children with CHD were collected. The mutations in coding regions 1 and 2 of the NKX2.5 gene, and coding regions 4, 5, and 8 of the TBX5 gene were examined by polymerase chain reaction (PCR) and DNA sequencing.</p><p><b>RESULTS</b>An A-to-G mutation at nucleotide 63 (c.63A>G) in coding region 1 of the NKX2.5 gene was found in both IVF and naturally conceived children with CHD. There were no significant differences in genotype and allele frequencies at c.63A>G locus of the NKX2.5 gene between the two groups. No mutations were detected in coding region 2 of the NKX2.5 gene and coding regions 4, 5 and 8 of the TBX5 gene.</p><p><b>CONCLUSIONS</b>There is no difference in NKX2.5 and TBX5 gene mutations between IVF and naturally conceived children with CHD. Therefore, it is presumed that assisted reproductive technology may not lead to mutations in the NKX2.5 and TBX5 genes.</p>

Value of combined measurement of intestinal fatty acid-binding protein and fecal calprotectin in diagnosis of necrotizing enterocolitis in full-term neonates / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the value of combined measurement of intestinal fatty acid-binding protein (I-FABP) and fecal calprotectin (FC) in the diagnosis of necrotizing enterocolitis (NEC) in full-term neonates.</p><p><b>METHODS</b>A total of 36 full-term neonates with NEC (case group) and 39 neonates without digestive system diseases (control group) were enrolled as study subjects. ELISA was used to measure the serum I-FABP level and fecal FC level, and the clinical value of I-FABP combined with FC in the diagnosis of NEC was evaluated.</p><p><b>RESULTS</b>The case group had significantly higher I-FABP and FC levels than the control group (P<0.05). In the case group, serum I-FABP level was positively correlated with fecal FC level (r=0.71, P<0.05). In the diagnosis of NEC, I-FABP alone, FC alone, and I-FABP/FC combination had sensitivities of 83.3%, 81.5%, and 79.5%, specificities of 72.5%, 75.8%, and 86.3%, and areas under the ROC curve (AUCs) of 0.82, 0.81, and 0.88. The combined measurement showed significantly higher specificity and AUC than single measurement (P<0.05).</p><p><b>CONCLUSIONS</b>Children with NEC have significant increases in I-FABP and FC levels, and there is a correlation between them. Combined measurement of I-FABP and FC can increase the specificity of the diagnosis of NEC.</p>

Neonatal complications and birth defects in infants conceived by in vitro fertilization / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the survival quality of infants conceived by in vitro fertilization (IVF) and to identify the factors that cause birth defects and neonatal complications in IVF infants.</p><p><b>METHODS</b>The study included 150 IVF infants (IVF group) and 200 naturally conceived infants (control group). Indicators such as birth situation, gestational disease, birth defects, and neonatal complications were compared between groups. The influencing factors for birth defects and neonatal complications were analyzed by non-conditional logistic regression analysis.</p><p><b>RESULTS</b>Compared with the control group, the IVF group had increased incidences of twin pregnancy and low birth weight (P<0.01) but decreased average birth weight (P<0.05). In the IVF group, the mother's age was elder, with higher incidence of cesarean section, premature rupture of membranes, and pregnancy complications, as compared with the control group (P<0.05). There was no significant difference in the incidence of birth defects between the two groups (P>0.05). The IVF group had higher incidence rates of low birth weight and neonatal scleroderma (P<0.05), with a longer hospital stay (P<0.01), as compared with the control group. The non-conditional logistic regression analysis indicated that IVF, prematurity, twin pregnancy, and pregnancy complications were risk factors for low birth weight (P<0.05).</p><p><b>CONCLUSIONS</b>There is no significant difference in the incidence of birth defects between IVF and naturally conceived infants. However, IVF infants have higher incidences of twin pregnancy and low birth weight, with a longer hospital stay, as compared with naturally conceived infants. Natural conceiving, avoiding prematurity, twin pregnancy, and pregnancy complications will reduce the incidence of low birth weight.</p>

Changes in human umbilical vein endothelial cells induced by endothelial nitric oxide synthase traffic inducer / 华中科技大学学报（医学）（英德文版）

This study investigated the changes in human umbilical vein endothelial cells (HUVECs) induced by overexpression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) and its role in cellular injury. Recombinant NOSTRIN-expressing and empty vectors were transfected into cultured HUVECs, and factor VIII-related antigen was examined by using immunohistochemical analysis. Growth curves were generated for both transfected and untransfected cells and these indicated that the proliferative ability of cells overexpressing NOSTRIN was significantly decreased. The expression of NOSTRIN and eNOS proteins was detected by using Western blot analysis, endothelial NOS (eNOS) activity was assayed by using spectrophotometry, and NO2 (-)/NO3 (-) levels were measured using nitrate reductase. Immunohistochemical analysis demonstrated that all groups expressed NOSTRIN in the plasma membrane and cytoplasm, and Western blot analysis confirmed that NOSTRIN levels were significantly higher in cells transfected with the NOSTRIN plasmid (P<0.01). The activity of eNOS and the levels of NO2 (-)/NO3 (-) were significantly decreased in NOSTRIN overexpressing cells as compared with empty vector and untransfected cells (P<0.01 and P<0.01, respectively). Morphological and ultrastructural changes were observed under light and electron microscopy, and it was found that NOSTRIN-overexpressing cells were elongated with deformities of the karyotheca, injury to the plasma membrane, increased lipids in the cytoplasm, and shortened microvilli. This study showed that overexpression of NOSTRIN had a significant effect on eNOS activity in HUVECs and resulted in significant cellular damage.

Changes in human umbilical vein endothelial cells induced by endothelial nitric oxide synthase traffic inducer / 华中科技大学学报（医学）（英德文版）

This study investigated the changes in human umbilical vein endothelial cells (HUVECs) induced by overexpression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) and its role in cellular injury. Recombinant NOSTRIN-expressing and empty vectors were transfected into cultured HUVECs, and factor VIII-related antigen was examined by using immunohistochemical analysis. Growth curves were generated for both transfected and untransfected cells and these indicated that the proliferative ability of cells overexpressing NOSTRIN was significantly decreased. The expression of NOSTRIN and eNOS proteins was detected by using Western blot analysis, endothelial NOS (eNOS) activity was assayed by using spectrophotometry, and NO2 (-)/NO3 (-) levels were measured using nitrate reductase. Immunohistochemical analysis demonstrated that all groups expressed NOSTRIN in the plasma membrane and cytoplasm, and Western blot analysis confirmed that NOSTRIN levels were significantly higher in cells transfected with the NOSTRIN plasmid (P<0.01). The activity of eNOS and the levels of NO2 (-)/NO3 (-) were significantly decreased in NOSTRIN overexpressing cells as compared with empty vector and untransfected cells (P<0.01 and P<0.01, respectively). Morphological and ultrastructural changes were observed under light and electron microscopy, and it was found that NOSTRIN-overexpressing cells were elongated with deformities of the karyotheca, injury to the plasma membrane, increased lipids in the cytoplasm, and shortened microvilli. This study showed that overexpression of NOSTRIN had a significant effect on eNOS activity in HUVECs and resulted in significant cellular damage.

Association of HAb18G with clinicopathologic features and prognosis in non-small cell carcinoma of lung / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the association between HAb18G expression, tumor parameters, metastatic potential and prognosis in non-small cell lung carcinoma (NSCLC).</p><p><b>METHODS</b>Immunohistochemical study for HAb18G protein using SP methods was carried out in 144 cases of NSCLC. Nineteen cases of benign lung lesions and 41 cases of normal lung tissue were used as controls. The intensity (positive unit/PU) of HAb18G expression was assessed quantitatively by image analysis software. The results were correlated with tumor parameters, metastatic potential and follow-up data.</p><p><b>RESULTS</b>The intensity of HAb18G protein expression was significantly higher in NSCLC than that in controls (P = 0.000). In squamous cell carcinomas and adenocarcinomas, the expression of HAb18G protein in well-differentiated tumors was lower than that in moderately to poorly differentiated tumors (P = 0.001). Tumors of TNM stage IV had stronger expression than tumors of lower stages (P = 0.000). HAb18G PU was greater in tumors with lymph node metastasis than those without nodal metastasis (P = 0.045). The PU value of tumors with maximal diameter greater than 5 cm was higher than that of the smaller tumors (P = 0.000). It was also higher in male than in female patients (P = 0.046). There was no association between HAb18G protein expression and age of patients, history of smoking, tumor types and gross morphology (P > 0.05). The five-year survival rate in cases with low HAb18G protein expression was higher than that in cases with high expression (P = 0.006). Univariate analysis indicated that patients with high HAb18G protein expression carried a poor prognosis (P = 0.007). Multivariate analysis showed that expression of HAb18G protein was an independent prognostic factor in patients with NSCLC (P = 0.032, relative risk 3.962).</p><p><b>CONCLUSIONS</b>HAb18G protein expression is associated with tumor progression and prognosis. It may represent a useful biomarker for prognostic evaluation.</p>

A study on population-based prenatal screening and diagnosis of Down's syndrome in Jiangsu province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To screen and diagnose Down's syndrome during mid-term pregnancy to reduce the number of babies with Down's syndrome.</p><p><b>METHODS</b>With the multi-level of stratified cluster sampling, twenty thousand and eight hundred and three women at 15-20 weeks gestation were screened by maternal serum AFP and beta-hCG using the time resolved fluoroimmunoassay (TRFIA). Then the screened high-risk women were diagnosed by amniocentesis, cell culture and chromosome analyses. The born children were diagnosed by follow-up and peripheral blood chromosome analyses.</p><p><b>RESULTS</b>Six fetuses were diagnosed by serum screening and amniotic fluid chromosome analyses, and 3 born children were diagnosed by follow-up and peripheral blood chromosome analyses. Nine cases of Down's syndrome were detected in total, with the positive prenatal screen rate being 67% (6/9).</p><p><b>CONCLUSION</b>The prenatal screening and diagnosis can reduce the birth of Down's syndrome patients and improve the population quality. However, the diagnosis accuracy still needs to be improved to further reduce the false negative rate and prevent misdiagnosis.</p>

TMSG-1 and its roles in tumor biology / 癌症

TMSG-1 is a newly discovered tumor metastasis suppressor gene, which plays important roles in promoting apoptosis and inhibiting invasion and metastasis of tumor cells. The inhibitory function of TMSG-1 in tumor cells may be related to vacuolar H+-ATPase and ceramide, but the underlying mechanism remains unknown. Studies on TMSG-1 are limited worldwide, and only a research group in Shanghai and our group have recently studied on it. As a new research field, the function of TMSG-1 remains to be explored. This review discusses the discovery of TMSG-1, structure of its encoded protein, its roles and possible mechanism in inhibiting tumor invasion and metastasis.

HPLC-MS/MS method for determination of sodium cromoglycate concentration in human plasma and its pharmacokinetics / 药学学报

The study established an HPLC-MS/MS method for determining the concentrations of sodium cromoglycate in human plasma and evaluated the pharmacokinetics of nasal drops and nasal spray. A C18 column was used to separate sodium cromoglycate in plasma with a mobile phase of a mixture of ammonium-methanol (involves 50% acetonitrile) (15:85) at a flow rate of 0.4 mL x min(-1). Electronic spray ionization (ESI) and multiple-reaction monitoring (MRM) were used for the determination of sodium cromoglycate in human plasma. The linear range of the standard curve of sodium cromoglycate was from 0.3 to 20 ng x mL(-1), and the minimum concentration of detection was 0.3 ng x mL(-1). The extraction recovery was more than 94.1%, intra-day and inter-day RSD were less than 14.3%. After a single dose of sodium cromoglycate, the main pharmacokinetic parameters of nasal spray and nasal drops were as follows, T(1/2)(1.82 +/- 0.54) h, (1.59 +/- 0.52) h; Tmax (0.47 +/- 0.12) h, (0.44 +/- 0.15) h; Cmax, (9.79 +/- 4.66) ng x mL(-1), (10.88 +/- 4.05) ng x mL(-1); AUC(0-5 h)(11.52 +/- 3.46) ng x mL(-1) x h x h, (12.63 +/- 4.23) ng x mL(-1) x h, Fr(93.6 +/- 13.8)%. The method is sensitive, rapid and accurate. It is suitable for therapeutic drug monitoring and human pharmacokinetic study of sodium cromoglycate.