Clinical treatment outcomes and their changes in extremely preterm twins: a multicenter retrospective study in Guangdong Province, China / 中国当代儿科杂志

OBJECTIVES@#To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China.@*METHODS@#A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups.@*RESULTS@#Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05).@*CONCLUSIONS@#There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.

Review on Mechanism of Multi-target Intervention of Traditional Chinese Medicine on Metabolic Syndrome / 中国实验方剂学杂志

The pathogenesis of metabolic syndrome （MS） includes insulin resistance （IR）， central obesity， chronic low-grade inflammation， oxidative stress， endoplasmic reticulum stress， elevated free fatty acid levels， intestinal flora imbalance， renin angiotensin system abnormality， and autophagy activity deficiency， etc. Most researchers believe that IR plays a central role in the pathogenesis of MS， and abdominal obesity is an important initial factor of MS. According to the incidence and clinical characteristics， MS is classified as "obesity" "pidan" " abdominal fullness " and other diseases. It is said that the pathogenesis of MS is related to the deficiency of spleen and kidney， the formation of phlegm， turbidity， blood stasis and other pathological products， which damage the body's functions of qi， blood， yin and yang. Traditional Chinese medicine （TCM） has unique advantages in treating MS based on the holistic view and syndrome differentiation concept. It has multi-level， multi-target and multi-channel treatment characteristics. It can intervene insulin signal transduction， regulate adipocyte factor secretion level， relieve oxidative stress and endoplasmic reticulum stress response， regulate intestinal flora and renin angiotensin system， reduce free fatty acid level and regulation Autophagy and other ways to improve chronic low-grade inflammation and IR status， and then comprehensive prevention and treatment of MS and its complications. However， the following problems still exist：lack of high-quality randomized controlled clinical research and large sample real-world research， clinical unified diagnosis and treatment standard has not yet formed， lack of genetic animal model in basic research， relatively single signal pathway and target of experimental research， and difficulty in timely formation of clinical transformation of scientific research achievements. Therefore， we should make full use of modern scientific and technological means to carry out systematic and standardized multicenter， large sample， high-quality randomized controlled trials or real-world research， we should prepare perfect animal models， focus on the crosstalk relationship between multiple related cell signaling pathways， and actively explore the potential relationship between signaling pathways and prescription compatibility， so as to actively promote basic scientific research achievements Clinical practice may be the key research direction in the prevention and treatment of MS in TCM.

Effect of Huanglian Wendantang in Treatment of Metabolic Syndrome / 中国实验方剂学杂志

Metabolic syndrome (MS) is a pathological condition characterized by central obesity, insulin resistance, hypertension, and hyperlipidemia. With the increase of poor dietary habits and lifestyles in modern society, especially the poor living habits of sedentariness and less movement, the prevalence of MS has increased year by year. According to relevant data, the number of MS patients worldwide will reach about 2.568 billion by 2040, which will seriously endanger human life and health. Huanglian Wendantang, as a famous traditional Chinese medicine prescription for clearing away heat and drying dampness, regulating Qi and resolving phlegm, and benefiting the stomach and gall, has been proved to have significant pharmacological effects in lowering blood fat, reducing blood sugar and resisting inflammation by modern pharmacological studies, and widely used in the treatment of metabolic diseases, cardiovascular diseases and other systemic diseases. In recent years, a large number of studies have proved that Huanglian Wendantang has a significant effect on MS. In terms of clinical efficacy, it could significantly improve the pathological state of obesity, dyslipidemia, abnormal glucose metabolism and hypertension in MS patients. Meanwhile, it could also interfere with the inflammatory state, prethrombotic state, abnormal vascular regulation and other potential risk factors in the body, with a high safety and fewer side effects. In terms of experimental study, it could enhance the insulin sensitivity, and improve the insulin resistance of MS animal models and cell models through interventions in insulin signal transduction, inflammatory response, and antioxidant stress. By retrieving PubMed, CNKI, Weipu, Wanfang and other databases, the author summarized the study reports of Huanglian Wendantang on MS in recent years in three aspects: theoretical study, clinical efficacy study and experimental mechanism study, in the expectation of provide some scientific references for in-depth study of the mechanism of Huanglian Wendantang in treating MS and the development and clinical promotion of the prescription.

Serum metabolomics of preterm and full-term infants based on gas chromatography-mass spectrometry / 中国当代儿科杂志

OBJECTIVE@#To study the features of serum metabolites in preterm infants based on gas chromatography-mass spectrometry (GC-MS), and to find differentially expressed metabolites in the serum of preterm infants.@*METHODS@#Serum samples were collected from 19 preterm infants and 20 full-term infants before feeding. GC-MS was used to measure metabolic profiles, and the metabolic features of 397 serum metabolites in preterm infants were analyzed.@*RESULTS@#There was a significant difference in serum metabolic features between the preterm and full-term infants before feeding. There were significant differences between the full-term and preterm infants in the levels of metabolites such as O-phosphonothreonine, digicitrin, tannic acid, and fructose-1,6-diphosphate (P<0.01), suggesting that the above differentially expressed metabolites were highly differentiated between the preterm and full-term infants. Most differentially expressed metabolites were involved in the metabolic pathways such as ABC transporters, β-alanine and pyrimidines and were correlated with some clinical parameters (albumin and total bilirubin) (P<0.05).@*CONCLUSIONS@#There is a significant difference in serum metabolites between preterm and full-term infants before feeding. Metabolomics plays an important role in improving metabolic disorders and exploring metabolism-related diseases in preterm infants.

Human umbilical cord mesenchymal stem cells derived from Wharton's jelly differentiate into insulin-producing cells in vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Islet transplantation is an effective way of reversing type I diabetes. However, islet transplantation is hampered by issues such as immune rejection and shortage of donor islets. Mesenchymal stem cells can differentiate into insulin-producing cells. However, the potential of human umbilical cord mesenchymal stem cells (huMSCs) to become insulin-producing cells remains undetermined.</p><p><b>METHODS</b>We isolated and induced cultured huMSCs under islet cell culture conditions. The response of huMSCs were monitored under an inverted phase contrast microscope. Immunocytochemical and immunofluorescence staining methods were used to measure insulin and glucagon protein levels. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect gene expression of human insulin and PDX-1. Dithizone-staining was employed to determine the zinc contents in huMSCs. Insulin secretion was also evaluated through radioimmunoassay.</p><p><b>RESULTS</b>HuMSCs induced by nicotinamide and β-mercaptoethanol or by neurogenic differentiation 1 gene (NeuroD1) transfection gradually changed morphology from typically elongated fibroblast-shaped cells to round cells. They had a tendency to form clusters. Immunocytochemical studies showed positive expression of human insulin and glucagon in these cells in response to induction. RT-PCR experiments found that huMSCs expressed insulin and PDX-1 genes following induction and dithizone stained the cytoplasm of huMSCs a brownish red color after induction. Insulin secretion in induced huMSCs was significantly elevated compared with the control group (t = 6.183, P < 0.05).</p><p><b>CONCLUSIONS</b>HuMSCs are able to differentiate into insulin-producing cells in vitro. The potential use of huMSCs in β cell replacement therapy of diabetes needs to be studied further.</p>

Suppression to ameloblastoma xenografts of chicken embryo chorioallantoic membrane by tissue inhibitor of metalloproteinases-2 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore the invasiveness of xenografts on chicken embryo chorioallantoic membrane (CAM) after tissue inhibitor of metalloproteinase-2 (TIMP-2) gene transfection.</p><p><b>METHODS</b>Fresh ameloblastoma tissues were minced into 1-2 mm3 and transplanted on the CAM. There were three groups named as control group (Empt), plasma transfection group (Lipo), and TIMP-2 gene transfection group (P). The specimens were respectively investigated by microscope indifferent spots after implanting. The volume of the xenografts and the weight of xenografts in the termination time of the experiment were recorded. The invasiveness of xenografts was divided into four grades by pathological examination. Western blot analysis was performed to investigate matrix metalloproteinase-2 (MIMP-2) and TIMP-2 protein in xenografts.</p><p><b>RESULTS</b>Ameloblastoma tissues can survive on CAM and the tumor cells may invade it on 5-7 days after implanting. At 9 d after implanting, the invasiveness grades in P group were 7 in grade 0, 1 in grade 2, 0 in grade 3. The expression of TIMP-2 protein in P group was significantly higher than that in Empt group (P < 0.05). The expression of MMP-2 protein in P group was lower than that in Empt group (P < 0.05).</p><p><b>CONCLUSION</b>The xenotransplanted tumor model of human ameloblastoma on CAM was successfully established. The invasiveness of ameloblastoma xenografts was suppressed might be due to TIMP-2 gene transfection.</p>

Effect of cyclooxygenase-2 inhibitor on expression of matrix metalloproteinase-2 and invasion of tongue squamous cell carcinoma cell line Tca8113 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of celecoxib on adhesion, invasion, migration and matrix metalloproteinase-2(MMP-2) expression of tongue squamous cell carcinoma cell line Tca8113 cells.</p><p><b>METHODS</b>Following incubation with celecoxib, the Tca8113 cells were detected for cell adhesion and migration using cell adhesion assay and Boyden chamber invasion assay. The expression of cyclooxygenase-2 (Cox-2) protein in Tca8113 cells was tected with SP immunohistochemistry staining. The MMP-2 level in supernatant was detected with enzyme-linked immunosorbent assay. RESULTS; The adhesion and Boyden chamber invasion assays showed that, after treatment celecoxib, the ability of adhesion and migration of Tca8113 cells was significantly inhibited. Celecoxib could decrease the expression of Cox-2 protein in Tca8113 cell and decrease the MMP-2 level in supernatant.</p><p><b>CONCLUSION</b>Cox-2 inhibitor celecoxib can significantly inhibit the adhesion and migration of Tca8113 cells. The inhibitory effect on hesion and migration may be correlative with its effect on decrese of Cox-2 protein expression and secretion MMP-2 in Tca8113 cells.</p>

Celecoxib induces cell cycle arrest and apoptosis in human tongue squamous carcinoma cell line Tca8113 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of celecoxib on the cell cycle and apoptosis in human tongue squamous cell carcinoma (HTSCC) cell line Tca8113.</p><p><b>METHODS</b>Tca8113 cell line was cultured in the presence of different concentrations of celecoxib. MTT assay was used to measure cell survival rate, and flow cytometry performed to analyze the cell cycle distribution. Annexin V-FITC/PI staining was used to detect the early changes of apoptosis, and transmission electron microscope employed to observe the ultrastructural changes of the apoptotic cells.</p><p><b>RESULTS</b>Celecoxib inhibited the proliferation of Tca8113 cells in a dose-dependent manner, the effect of which was mediated by inducing cell cycle arrest mainly in G1/S phase. Flow cytometry and ultrastructural observation demonstrated an early to late stage changes of the apoptotic cells exposed to increased concentrations of celecoxib.</p><p><b>CONCLUSION</b>Celecoxib dose-dependently inhibits the proliferation of Tca8113 cells by causing cell cycle arrest and inducing apoptosis.</p>

High-performance liquid chromatography for determining celecoxib concentration in hamster tongue tissue after oral local application / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a high-performance liquid chromatography (HPLC)-based method for determining celecoxib concentration in the tongue tissue of hamsters.</p><p><b>METHODS</b>Celecoxib mixed with the matrix (final concentration of 6%) was smeared on the surface of the tongue mucosa of hamsters, and the concentration and absorption rate of celecoxib in the tongue tissue were determined by HPLC at 5, 10, 15, 30, 60, 90, 120 min after the application.</p><p><b>RESULTS</b>In this system, the retention time of celecoxib was 4.4 min. Celecoxib concentration showed a good linear range within 25-800 microg/L (R2=0.9991, n=6), with the detection limit for celecoxib of 10 g/L (S/N=3). The extraction recoveries and method recoveries for celecoxib were 83.75%-90.01% and 91.98%-99.07%, respectively. The inter-day RSDs were 2.15%, 3.16% and 3.67%, and intra-day RSDs were 3.40%, 4.56% and 4.42%, respectively. The concentration of celecoxib in hamster tongue tissue within the first 120 min ranged from 0.685-/+0.019 microg/g to 3.168-/+0.143 g/g, reaching the peak level at 15 min.</p><p><b>CONCLUSION</b>Celecoxib can be rapidly absorbed through the tongue mucosa to reach a high concentration in the tongue tissue, indicating the possibility of oral COX-2 inhibitors to prevent oral cancer and precancerous lesions.</p>

Celecoxib enhances the lethal effects of bleomycin in human tongue squamous carcinoma cell line Tca8113 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the role of COX-2 inhibitor celecoxib in enhancing the lethal effects of bleomycin in Tca8113 cell line.</p><p><b>METHODS</b>Tca8113 cells were treated with different concentrations of celecoxib and bleomycin for 24, 48, 72 h. Methyl thiazolyl tetrazolium assay was used to calculate cell growth inhibition rate and Jin Zheng Jun's method was used to evaluate the interaction of celecoxib and bleomycin on Tca8113 cells. Flow cytometry was used to evaluate the effects of combined use of celecoxib and bleomycin on cell cycle progress and apoptosis.</p><p><b>RESULTS</b>Low dose of celecoxib (10 micromol/L, < IC(50)) combined with bleomycin showed synergism or additive lethal effect on Tca8113 cell line. Celecoxib could notably enhance the inhibitory effect of bleomycin on Tca8113 cells by blocking cell cycle progress and thus resulting in the increasing G(0)/G(1) cells [(60.93 +/- 0.32)%] distribution and inducing apoptosis [(1.87 +/- 0.11)%].</p><p><b>CONCLUSIONS</b>Low doses of celecoxib could significantly enhance the lethal effect of bleomycin on Tca8113 cells by inhibiting cell growth and proliferation through blocking cell cycle progress and inducing apoptosis. The ways of these interactions on inhibiting Tca8113 cell growth were synergistic or/and additive.</p>

Inhibitory effects of COX-2 inhibitor on migration of human tongue squamous cell carcinoma Tca8113 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of COX-2 inhibitor celecoxib in inhibiting the migration of human tongue squamous cell carcinoma Tca8113 cells.</p><p><b>METHODS</b>The effects of celecoxib on Tca8113 cell migration were tested using scrape motility assay, cell-matrix adhesion assay and Boyden chamber motility assay.</p><p><b>RESULTS</b>Following a 24-hour incubation with 10 and 20 micromol/L celecoxib, the migration of Tca8113 cells was significantly decreased (Plt;0.05). Celecoxib treatment for 24 h also resulted in significantly decreased adhesion of Tca8113 cells on Fn-coated surface in a dose-dependent manner.</p><p><b>CONCLUSION</b>COX-2 inhibitor celecoxib can inhibit Tca8113 cell migration, the mechanism of which awaits further investigation.</p>

Effect of Celecoxib on cycloxygenase-2 expression and inducing apoptosis in Tca8113 cell lines / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe whether Celecoxib could inhibit the growth, regulate the expression of COX-2 and induce apoptosis of Tca8113 cells.</p><p><b>METHODS</b>Tca8113 cells were incubated with different concentrations of Celecoxib for 24, 48 and 72 h, and MTT was used to calculate growth inhibition rate. The expression of COX-2 protein and mRNA in Tca8113 cells was detected with SP immunohistochemistry staining and fluorescent quantitative real-time RT-PCR. Morphology of apoptosis cells was observed by fluorescence microscopy, and Annexin V-FITC/PI double labeling method was employed to detect early stage cell apoptosis.</p><p><b>RESULTS</b>COX-2 protein was strongly expressed in Tca8113 cells and was suppressed by Celecoxib. The growth and proliferation of Tca8113 cells treated with Celecoxib were inhibited in a dose-dependent manner. Celecoxib treatment resulted in significant increase in apoptosis and early apoptotic rate. Fluorescent quantitative real-time RT-PCR results showed no significant effet on regulating expression of COX-2 mRNA.</p><p><b>CONCLUSION</b>Celecoxib shows a significant effect on inhibiting expression of COX-2 in Tca8113 cells, this is probably related to growth inhibition and inducing apoptosis of Tca8113 cells.</p>

Effects of celecoxib on PGE2 synthesis and COX-2 and VEGF-C mRNA expression in Tca8113 cell lines / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of the selective cyclooxygenase-2 (COX-2) inhibitor celecoxib on prostaglandin E(2) (PGE2) release and vascular endothelial growth factor C (VEGF-C) and COX-2 mRNA expression in Tca8113 cell lines.</p><p><b>METHODS</b>MTT assay was used to analyze the proliferation of Tca8113 cells. The PGE2 level was detected with enzyme-linked immunosorbent assay (ELISA), and the expressions of COX-2 and VEGF-C mRNA were examined with RT-PCR.</p><p><b>RESULTS</b>Celecoxib could induce inhibitory effects on the growth and PGE2 release in Tca8113 cells. The RT-PCR results showed that celecoxib significantly down-regulated the expression of VEGF-C mRNA, but produced a weak effect on COX-2 mRNA expression.</p><p><b>CONCLUSION</b>The inhibitory effect of celecoxib on Tca8113 cell growth and the expressions of VEGF-C and COX-2 may be related to the release of PGE2.</p>

Expression of COX-2 and VEGF-C in squamous cell carcinoma of the tongue and its correlation to lymph node metastasis / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the coexpression of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor-C (VEGF-C) in human squamous cell carcinoma of the tongue (SCCT) and assess their correlations to neoangiogenesis and lymph node metastasis of the tumor.</p><p><b>METHODS</b>Tissue samples of primary SCCT and the metastatic lymph nodes were obtained from 46 patients undergoing surgical resections of SCCT for immunohistochemical detection of COX-2 and VEGF-C expressions.</p><p><b>RESULTS</b>The over-expression rates of COX-2 and VEGF-C was 82.61% and 73.91% in the primary tumor, respectively, significantly higher than those in normal oral mucosa. A significant correlation was found between the expression scores of COX-2 and VEGF-C with a concordance rate of 82.61% (P<0.01) and a kappa value of 0.495 according to Cohen's kappa test (P<0.01). High expression of VEGF-C, but not COX-2, was correlated to the presence of lymph node metastasis (P<0.05). The mean microvessel density was significant higher in tumors with strong positivity for COX-2 or VEGF-C than in tumors with only weak or negative expressions (P<0.001).</p><p><b>CONCLUSION</b>The expressions of COX-2 and VEGF-C are closely correlated in SCCT, and may have clinical value for assessing the prognosis, metastasis and invasion of SCCT, but the mechanism of these proteins in carcinogenesis in SCCT needs further investigation.</p>

Expression of HSV-1 ICP0 Antigen Peptide in Prokaryotic Cells and Preparation of Specific Antibody / 中国病毒学

As an immediate-early protein of herpes simplex virus, infected-cell polypeptide 0 (ICP0) exhibits complicated interactions with host cells, and its regulatory function on gene expression is of great importance. Since the ICP0 encoding sequence contains many rare codons which are absent in E.coli, and ICP0 is highly unstable in prokaryotic cells, expression of entire ICP0 in prokaryotic cells has never been reported. In order to further investigate the function of ICP0, a recombinant plasmid was constructed by subcloning a cDNA fragment encoding an amino-terminal of 105 residues of the ICP0 protein into pGEX-5x-1 vector. The resulting GST-105 fusion antigen peptide was expressed with high efficiency in E.coli. Antibodies prepared after the immunization of mice with purified fusion protein can recognize not only the denatured ICP0 protein, but also the native ICP0 protein with normal biological conformation.

Prokaryotic Expression and Acetylation Assays of Histone Acetyltransferase PCAF / 中国生物工程杂志

P300/CBP-associated factor(PCAF),an important member of histone acetyltransferase family(HATs) within eukaryotic cells,is capable of inducing the acetylation of histone,promoting the transcription of specific genes and involving in many biological effects.In the present study,full-length cDNA of PCAF was inserted into plasmid pGEX-5x-1,then the soluble protein GST-PCAF was expressed in E.coli BL21(DE3) after the optimization of inducing conditions.The recombinant protein was further purified with affinity chromatography and tested the activity by in vitro acetylation assays.High efficient PCAF protein produced by this method could serve for the study on the role of PCAF in gene regulation and the interaction between PCAF and other proteins.