[Whole-course management strategies for gastroesophageal reflux disease: an evidence mapping study based on clinical practice guidelines].

Gastroesophageal reflux disease (GERD) is one of the most common digestive diseases with high incidence, complicated clinical symptoms, difficulties in standard treatment, and heavy medical burden. At present, some GERD-relevant clinical practice guidelines (CPGs) have been issued by different countries and academic organizations, but some recommendations were inconsistent, which has caused some problems for the current clinical whole-course management of GERD. To summarize the relevant evidence among the CPGs on GERD and formulate the whole- course management strategies, we included GERD-relevant CPGs published or updated after 2010 by searching websites of guidelines, relevant professional societies, and electronic databases. We extracted the recommendations and summarized the evidence from the aspects of symptoms, epidemiology, diagnosis and treatment, which was presented in the form of evidence mapping. We included 24 CPGs, including three in Chinese and 21 in English. The clinical practice management strategies of GERD were formulated based on the evidence from the aspects of clinical symptoms, diagnostic methods, medical treatment, anti-reflux surgery and endoscopic treatment, psychological treatment, and traditional Chinese medicine treatment.

[Study on the relationship between ABCB1 gene polymorphism and hemorrhage risk after thrombolysis of cerebral ischemic stroke in Shangqiu area].

Using a cross-sectional study, 246 patients with hemorrhage and transformation after cerebral ischemic stroke(CIS) thrombolysis who were admitted to Shangqiu First People's Hospital, Shangqiu Municipal Hospital, and Shangqiu Liangyuan Traditional Chinese Medicine Hospital from March 2018 to May 2020 were selected as the observation group, 246 patients with no hemorrhage transformation after CIS thrombolysis during the same period were selected as the control group with a ratio of 1∶1. Polymerase chain reaction and pyrosequencing methods were used to detect the single nucleotide polymorphisms of the two groups of ABCB1 genes. The frequency distribution of each genotype of the two groups of ABCB1 gene polymorphism sites was counted. The conditional logistic regression equation was used to analyze the CIS after thrombolysis. Related influencing factors of hemorrhage transformation, and compare the single nucleotide polymorphisms of ABCB1 gene in patients with different prognosis in the observation group. The results showed that the CC genotype frequency of rs1045642 in the observation group was 34.55% higher than that of the control group 25.02%, the CT genotype frequency was 12.20%, and the TT genotype frequency 3.25% was lower than that of the control group 14.63% and 9.35% (χ2=21.527, P<0.05); GG genotype frequency at rs2032582 locus in observation group was 17.89%, GT genotype frequency 21.54% was lower than control group 37.60%, 93.96%, TT genotype frequency 10.57% higher than control group 2.44%, the difference was statistically significant (χ2=80.427, P<0.05); TT genotype at rs1045642 is a protective factor for hemorrhage transformation, and TT genotype at rs2032582 is a risk factor for hemorrhage transformation (OR=2.903, P<0.05). The risk of bleeding after thrombolysis in CIS patients in Shangqiu area may be related to the TT genotype at the ABCB1 rs1045642 locus and the TT genotype at the rs2032582 locus.

Responses of the soil fungal communities to the co-invasion of two invasive species with different cover classes.

Soil fungal communities play an important role in the successful invasion of non-native species. It is common for two or more invasive plant species to co-occur in invaded ecosystems. This study aimed to determine the effects of co-invasion of two invasive species (Erigeron annuus and Solidago canadensis) with different cover classes on soil fungal communities using high-throughput sequencing. Invasion of E. annuus and/or S. canadensis had positive effects on the sequence number, operational taxonomic unit (OTU) richness, Shannon diversity, abundance-based cover estimator (ACE index) and Chao1 index of soil fungal communities, but negative effects on the Simpson index. Thus, invasion of E. annuus and/or S. canadensis could increase diversity and richness of soil fungal communities but decrease dominance of some members of these communities, in part to facilitate plant further invasion, because high soil microbial diversity could increase soil functions and plant nutrient acquisition. Some soil fungal species grow well, whereas others tend to extinction after non-native plant invasion with increasing invasion degree and presumably time. The sequence number, OTU richness, Shannon diversity, ACE index and Chao1 index of soil fungal communities were higher under co-invasion of E. annuus and S. canadensis than under independent invasion of either individual species. The co-invasion of the two invasive species had a positive synergistic effect on diversity and abundance of soil fungal communities, partly to build a soil microenvironment to enhance competitiveness of the invaders. The changed diversity and community under co-invasion could modify resource availability and niche differentiation within the soil fungal communities, mediated by differences in leaf litter quality and quantity, which can support different fungal/microbial species in the soil.

RESEARCH ARTICLE Molecular cloning and expression analysis of a matrix Gla protein gene in the spinyhead croaker, Collichthys lucidus.

The matrix Gla (gamma-carboxyglutamic acid-rich) protein (MGP), a vitamin K-dependent and Gla-containing protein, is a calcification inhibitor that mainly functions in tissue calcification and mineralization. In this study, we obtained the complete cDNA sequence of MGP from the spinyhead croaker (Collichthys lucidus), which we named Cl-MGP. Cl-MGP was 923 bp long with a 384-bp open reading fragment that encoded 127 amino acids. The predicted MGP protein sequence contained a 19-residue hydrophobic signal peptide, suggesting that it possesses secretory characteristics. The Gla domain and the invariant unit ErraEtCedyspC, which has been identified in all known vitamin K-dependent vertebrate proteins, were highly conserved in Cl-MGP, suggesting that it uses the same mechanism to function as the known proteins. An alignment analysis revealed that Cl-MGP had the highest identity with Larimichthys crocea (93%), which had lost five amino acid residues in the C-terminal. A quantitative real-time polymerase chain reaction revealed that Cl-MGP expression was highest in the gill, followed by the cholecyst and spleen, with almost no expression in the blood, muscle, or testes. The high Cl-MGP expression in the gill is similar to that observed in other fish species, but the relatively high expression found in the cholecyst and spleen is not seen in all species. Future studies should investigate the tissue distributions of both mRNA and proteins in different species, in order to understand the function and evolution of MGP in different species.

[Relationship between full flowering stages of anemophilous and ligneous plants and consultation rate of allergic rhinitis].

Objective:The aim of this study is to investigate the climatic changes on spring phenology impacting on the consultation rate of patients with allergic rhinitis.Method:We analyzed the periods of full flowering stages of 15 kinds of flowers in Beijing from 2013 to 2014,and the number of patients with allergic rhinitis in the outpatient clinic of Otolaryngology Department of Peking University Third Hospital in the same time.The correlation test, analysis of variance and rank sum test were used to investigate the inherent laws and associations between the full flowering stages and the ratio of patients with allergic rhinitis.Result:①Between 2013 and 2014,the early and middle April were the full flowering stages of Platycladus orientalis,Salix matsudana, Fraxinus chinensis,Elms and Birches.The number of patients with allergic rhinitis in outpatient clinic elevated obviously in this period.②There were overlaps of full flowering stages among different plants.During periods in which three or more kinds of pollens in full flowering stages,the consultation rate of patients with allergic rhinitis is higher than less overlaping flower period.The difference was statistically significant(F value were -2.281,-2.964 respectively, P <0.05).Also,there were obvious positive linear correlations between the number of overlap of different pollens and the consultation rate of patients with allergic rhinitis in in 2013 and 2014(r=0.957, 0.964 respectively).Conclusion:During the overlaping flower periods,there was a higher consultation rate of patients with allergic rhinitis.In prevention and treatment of allergic rhinitis,the overlapping effect of full flowering stages of different pollens should be taken into account.Upgrade the alarm level will reduce the incidence of AR and the degree of allergic rhinitis episodes.

Molecular cloning and mRNA expression of a hepcidin gene from the spinyhead croaker, Collichthys lucidus.

Antimicrobial peptides are important components that participate in host innate immune activities and play crucial roles in host defense against microbial invasion. Hepcidin is an antimicrobial peptide and iron-regulatory molecule that primarily functions in the liver. In the present study, we first obtained a full-length cDNA sequence of hepcidin and its corresponding genomic DNA sequence from Collichthys lucidus using RT-PCR and rapid amplification of cDNA ends (RACE), and then analyzed these sequences using bioinformatics software. The results showed that C. lucidus hepcidin (CL-hepc) possesses two introns and three exons in the genomic DNA, with a length of 816 bp. The open reading frame was 264 bp, encoding an 87 amino acid peptide, and with high similarity (88.89%) to 83416593 Larimichthys crocea (ABC18307) and relatively low similarity (47.73%) to 158358729 L. crocea (ABY84845.1). The pre-peptide contained a signal peptide (28 amino acids), a prodomain (34 amino acids), and a mature peptide (25 amino acids). The predicted 25 amino acid hepcidin mature peptide included 8 conserved cysteine residues. Quantitative real-time reverse transcription-PCR analysis revealed specific expression patterns of CL-hepc, with the highest expression observed in the liver, relatively low expression observed in the gill and spleen, and almost no expression detected in other tissues analyzed. In conclusion, we identified a hepcidin from C. lucidus that has common expression patterns with other hepcidins. However, as this hepcidin is inconsistent with two other hepcidins from L. crocea in terms of the phylogenetic tree, the presence of another hepcidin gene warrants further investigation.

Black cohosh improves objective sleep in postmenopausal women with sleep disturbance.

OBJECTIVE: Sleep problems are prominent after menopause. The aim of our study was to look into the effect of black cohosh on both objective and subjective sleep in early postmenopausal women with sleep complaints. METHODS: We performed a randomized, double-blind and placebo-controlled research during a 6-month period. Forty-eight postmenopausal women aged 45-60 years with sleep disturbance were enrolled and received daily administration of either black cohosh or placebo. Polysomnography and the Pittsburg Sleep Quality Index (PSQI) were performed at the initiation and termination of the study, as well as the Menopause-specific Quality of Life questionnaire and estradiol and follicle stimulating hormone tests. Liver and renal functions and breast and pelvic ultrasound were set as safety measures, carried out every 3 months. RESULTS: Seventy-six women were interviewed, of whom 42 women completed the whole trial. Compared with placebo, black cohosh treatment led to significant polysomnographic changes, including increased sleep efficiency and decreased wake after sleep onset (WASO) duration, and tended to improve PSQI with a medium effect size. On average, 15.8% of WASO duration was reduced in the black cohosh group. Vasomotor and physical domains of life quality were improved compared with placebo. Safety measures did not yield any adverse event assigned to black cohosh. CONCLUSIONS: In early postmenopausal women with a major sleep complaint, black cohosh effectively improved sleep and might be a safe measure in managing menopausal sleep disturbance.

Molecular cloning and mRNA expression profiling of the first specific jasmonate biosynthetic pathway gene allene oxide synthase from Hyoscyamus niger.

In the endeavor to enhance the production of pharmaceutically valuable tropane alkaloids including hyoscyamine and scopolamine in Hyoscyamus niger, methyl jasmonate (MeJA) showed significant stimulation both in tropane biosynthetic pathway enzymes activities and tropane alkaloids yields. Therefore it was speculated that genetic engineering of jasmonate biosynthetic pathway might enhance the endogenous jasmonate concentration, followed by stimulating the production of tropane alkaloids. Herein a full-length cDNA encoding allene oxide synthase (AOS, EC 4.2.1.92), the first committed step enzyme in jasmonate biosynthetic pathway was reported (named HnAOS, GenBank accession: EF532599). HnAOS was a novel member of the cytochrome P450 (CYP74A) subfamily. Real-time quantitative PCR analysis showed that HnAOS mRNA accumulated mainly in stems, and responded significantly to wounding or methyl jasmonate.

Functional involvement of protein kinase C-betaII and its substrate, myristoylated alanine-rich C-kinase substrate (MARCKS), in insulin-stimulated glucose transport in L6 rat skeletal muscle cells.

AIMS/HYPOTHESIS: Insulin stimulates phosphorylation cascades, including phosphatidylinositol-3-kinase (PI3K), phosphatidylinositol-dependent kinase (PDK1), Akt, and protein kinase C (PKC). Myristoylated alanine-rich C-kinase substrate (MARCKS), a PKCbetaII substrate, could link the effects of insulin to insulin-stimulated glucose transport (ISGT) via phosphorylation of its effector domain since MARCKS has a role in cytoskeletal rearrangements. METHODS: We examined phosphoPKCbetaII after insulin treatment of L6 myocytes, and cytosolic and membrane phosphoMARCKS, MARCKS and phospholipase D1 in cells pretreated with LY294002 (PI3K inhibitor), CG53353 (PKCbetaII inhibitor) or W13 (calmodulin inhibitor), PI3K, PKCbetaII and calmodulin inhibitors, respectively, before insulin treatment, using western blots. ISGT was examined after cells had been treated with inhibitors, small inhibitory RNA (siRNA) for MARCKS, or transfection with MARCKS mutated at a PKC site. MARCKS, PKCbetaII, GLUT4 and insulin receptor were immunoblotted in subcellular fractions with F-actin antibody immunoprecipitates to demonstrate changes following insulin treatment. GLUT4 membrane insertion was followed after insulin with or without CG53353. RESULTS: Insulin increased phosphoPKCbetaII(Ser660 and Thr641); LY294002 blocked this, indicating its activation by PI3K. Insulin treatment increased cytosolic phosphoMARCKS, decreased membrane MARCKS and increased membrane phospholipase D1 (PLD1), a protein regulating glucose transporter vesicle fusion resulted. PhosphoMARCKS was attenuated by CG53353 or MARCKS siRNA. MARCKS siRNA blocked ISGT. Association of PKCbetaII and GLUT4 with membrane F-actin was enhanced by insulin, as was that of cytosolic and membrane MARCKS. ISGT was attenuated in myocytes transfected with mutated MARCKS (Ser152Ala), whereas overproduction of wild-type MARCKS enhanced ISGT. CG53353 blocked insertion of GLUT4 into membranes of insulin treated cells. CONCLUSIONS/INTERPRETATION: The results suggest that PKCbetaII is involved in mediating downstream steps of ISGT through MARCKS phosphorylation and cytoskeletal remodelling.

Corticosterone administration and dietary glucose supplementation enhance fat accumulation in broiler chickens.

1. The effects of exogenous corticosterone administration and glucose supplementation on energy intake, lipid metabolism and fat deposition of broiler chickens were investigated. 2. A total of 144 three-d-old male chickens were randomly assigned to one of the following 4 treatments for 7 d: a low energy diet (10.9 MJ ME/kg, 200 g/kg CP) with or without corticosterone (30 mg/kg diet) and drinking water supplemented with glucose (80 g/l) or saccharine (2 g/l, control). 3. Body weight (BW) gain and breast and thigh muscle yields (% body mass) were all significantly decreased by corticosterone treatment. The relative cumulative feed intake (RCFI) and relative ME intake (RMEI), rather than the feed (FI) or ME intake (MEI) were increased by corticosterone administration. Both feed efficiency (FE) and caloric efficiency (CE) were decreased by corticosterone administration. Corticosterone administration had no obvious effect on water consumption. 4. Glucose supplementation had no influence on BW gain and breast and thigh muscle yield (as % of body mass). FI or RCFI was decreased while MEI or RMEI was increased by glucose supplementation. FE was improved by glucose treatment, whereas CE was reduced. 5. Liver weight and abdominal, cervical and thigh fat deposits were all significantly increased by either corticosterone or glucose treatment. 6. Plasma concentrations of glucose, urate, triglyceride, non-esterified fatty acids (NEFA), very low density lipoprotein and insulin were all significantly increased by corticosterone treatment. Glucose supplementation had no obvious influence on any of the measured plasma parameters except for NEFA, which were significantly increased. 7. Lipoprotein lipase activities in either cervical or abdominal adipose tissues, rather than in thigh fat tissue, were significantly elevated by either glucose or corticosterone treatment.

[Molecular cloning and expression profile of a jasmonate biosynthetic pathway gene allene oxide cyclase from Hyoscyamus niger].

Hyoscyamus niger L. is a medicinal plant which produces a class of jasmonate-responsive pharmaceutical secondary metabolites named as tropane alkaloids. As a family of signaling phytohormones, jasmonates play significant roles in the biosynthesis of many plant secondary metabolites. In jasmonate biosynthetic pathway of plants, allene oxide cyclase (AOC, [...] EC 5.3.99.6 [...]) catalyzes the most important step. Here we cloned a cDNA from H. niger, named HnAOC (GenBank accession: AY708383), which was 1044 bp long, with a 747 bp open reading frame (ORF) encoding a polypeptide of 248 amino acid residues. Southern blot analysis indicated that it was a multi-copy gene. RT-PCR analysis revealed that the expression of HnAOC was regulated by various stresses and elicitors, with methyl-jasmonate showing the most prominent inducement. The characterization of HnAOC would be helpful for improving the production of valuable secondary metabolites by regulating the biosynthesis ofjasmonates.

Effects of tanshinone II-A sulfonate on adhesion molecule expression of endothelial cells and platelets in vitro.

AIM: To study the action of tanshinone II-A sulfonate (Tan) on adhesion molecule expression by cultured endothelial cells and platelets. METHODS: Tumor necrosis factor alpha (TNF-alpha)-induced ICAM-1 expression on the cell surface and endothelial adhesivity toward HL-60 cells were studied using human umbilical vein endothelial cells (HUVEC). Thrombin-induced expression of platelet P-selectin was studied using human blood platelets. Adhesion molecule expression on the cell surface was measured by flow cytometry. The number of HL-60 cells adhering to the HUVEC monolayer was determined by liquid scintillation spectroscopy. RESULTS: Pretreatment of HUVEC with TNF-alpha significantly enhanced ICAM-1 expression and increased HL-60 cells adhesion to HUVEC from 4.6% +/- 0.7% to 30% +/- 6%. Tan (25-200 mumol.L-1) inhibited the effects of TNF-alpha in a concentration-dependent manner. Tan also inhibited the increase of P-selectin expression of thrombin-activated platelets in a concentration-dependent manner. CONCLUSION: Tan inhibited expression of adhesion molecules (ICAM-1, P-selectin) in HUVEC and in human blood platelets.

[Effect of antisense c-erbB2 on biologic behaviour and chemotherapeutic drug sensitivity in human ovarian cancer cells].

OBJECTIVE: To explore the effects of antisense erbB2 on the biological behaviours and on chemotherapeutic drug sensitivity in human ovarian cancer cells. METHODS: A recombinant retroviral expression vector: pDOR-erbB-neo was constructed which contain the neomycin resistant gene and the erbB2 cDNA 3.8kb antisense fragment. pDOR-erbB-neo was introduced into human ovarian cancer cell line SKOV3 by lipofectin. The cells were selected with geneticin (G418). The cells transfected with pDOR-erbB-neo were named SKOV3-A2. RESULTS: Southern blot analysis confirmed the presence of the antisense erbB2 in transfected cells. Transfected cells showed no obvious changes in morphology. Compared with parental cells and with pDOR-neo transfected cells used as controls, the growth and DNA synthesis of antisense transfected cells, SKOV3-A2, were inhibited. Transfection with pDOR-erbB-neo rendered the cells significantly, more sensitive to chemotherapeutic drugs (5-fluorouracil, cisplatinum) than the parental cells. CONCLUSIONS: The effectiveness and the potentiality of antisense erbB2 in ovarian cancer gene therapy is demonstrated. c-erbB2 oncogene may be related to drug resistance in ovarian cancer cell.

Effects of Panax notoginseng saponins on posthypoxic cell damage of neurons in vitro.

AIM: To study cerebral protective mechanism of Panax notoginseng saponins (PNS). METHODS: Cultured neurons of chick embryo cerebral hemisphere were used as an in vitro system for investigating the effects of PNS. The hypoxic cell damage of neurons cultured were induced by NaCN. The levels of adenosine triphosphate (ATP) were determined with HPLC. PNS was added 30 min before, beginning or after hypoxia. RESULTS: PNS 50 and 100 mg L-1 retarded the break down of ATP of cultured neurons after 2-h hypoxia for 11.3 +/- 1.5 (P < 0.05) and 12.8 +/- 2.2 mumol/g protein (P < 0.01), respectively and accelerated the restoration of ATP during 30-min reoxygenation for 21.0 +/- 2.0 (P < 0.05) and 22.7 +/- 2.6 mumol/g protein (P < 0.01), respectively. PNS also reduced the release of creatine kinase (CK) from 75 +/- 8 kU L-1/g protein to 52 +/- 6 (P < 0.05) and 41 +/- 3 kU L-1/mg protein (P < 0.01), respectively and promoted the restoration of ATP of neurons 20 h after hypoxia when administered in the beginning of hypoxia from 13.0 +/- 0.9 mumol/g protein to 18.1 +/- 1.4 and 20.5 +/- 2.1 mumol/g protein (P < 0.01), respectively. PNS still promoted the restoration of ATP from 13.0 +/- 0.9 nmol/mg protein to 14.9 +/- 1.0 and 18.3 +/- 0.7 nmol/mg protein (P < 0.01), respectively and reduced (PNS 100 mg L-1) the CK release of neurons 20 h after hypoxia even when added in the recovery. CONCLUSION: The protection against hypoxic damage of PNS was related to improving energy metabolism, preserving the structural integrity of neurons.

[Experimental study on combined treatment in smoke inhalation injury].

Forty-two healthy dogs were randomly divided equally into a control group (CG) and a treated group (TG). All were inflicted with severe smoke inhalation injury and pulmonary was demonstrated. The dogs in TG were treated with injection of shenmai zhusheye, ketoprofen, anisodamine, sodium aesculin, hydrocortisone succinate, vitamin C and E, penicillin, amikacin, oxygen inhalation and airway suctioning after injury. The results showed that the increase in extravascular lung water volume, lung edema shadow in x-ray films, elevation of lung vascular resistance, carbonemia, hypoxemia, respiratory alkalosis, metabolic acidosis, dyspnea, dry and moist rales of the lungs, reduction of tidal volume, etc, were markedly improved in TG as compared with that of CG. It indicated that the pulmonary edema and lung dysfunction was markedly ameliorated in TG. The mortality was 19.1% in TG, which was significant lower than that of CG which was 47.6%.

[A preliminary study on pollinating bees for Amomum compactum Soland ex Maton].

This article deals with the species of bees pollinating Amomum compactum, along with the laws governing their growth and decline, and the pollinating effect of Nomia and Ceratina in the field.