Objective To observe the expression of ghrelin and growth hormone secretagogue receptor (GHSR) in pancreas of rats with acute necrotizing pancreatitis (ANP),and investigate the role of ghrelin in the pathogenesis of ANP.Methods Seventy SD rats were randomly divided into ANP group (n =35) and control group (n =35).ANP model was induced by retrograde injection of 4％ sodium taurocholate into the biliary and pancreatic duct.Rats in the control group underwent laparotomy with gentle pancreas manipulation only.At 3,6,12,24,48 h after ANP induction,the rats were sacrificed,and the serum level of amylase was determined,pathological changes in pancreatic tissue were routinely observed and scored.The expressions of ghrelin mRNA,protein and GHSR mRNA,protein in pancreas were evaluated by RT-PCR and Western blot.Results Serum amylase level began to increase at 3h after sodium taurocholate injection and reached the peak value at 6 h [(8244 ± 2950) U/L],which was significantly higher than that in control group (P ＜ 0.05).Pancreatic injuries was aggravated with time,the pathologic score at 24 h was (11.91 ± 1.31) score,which was significantly higher than (3.12 ± 1.60) score in the control group.The expressions of ghrelin mRNA and GHSR mRNA in pancreas of ANP group were increased gradually with time,and were significantly higher than those of control group at all time points,at 48 h,1.29 ±0.64 vs 0.58 ±0.05 and 0.94 ±0.16vs 0.19 ±0.03,P ＜ 0.05.The expressions of ghrelin protein and GHSR protein in pancreas of ANP group were significantly higher than that in control group at 12,24,48 h.at 48 h,3.05 ± 0.48 vs 2.18 ± 0.23 and 2.34 ± 0.32 vs 1.55 ± 0.10 (P ＜ 0.05).Conclusions The expressions of ghrelin mRNA,protein and GHSR mRNA,protein of pancreas are significantly increased in rats of ANP,and are associated with the severity of ANP.

Objective To investigate the clinical value of the Classification of acute pancreatitis2012.Methods Medical records and clinical data of patients with acute pancreatitis (AP) who were admitted to First Affiliated Hospital of Guangxi Medical University between October 2009 and September 2012 were retrospectively reviewed and analyzed.Patients were divided into mild acute pancreatitis (MAP),moderately severe acute pancreatitis (MSAP),and severe acute pancreatitis (SAP) groups according to the Classification of acute pancreatitis-2012.The number of improved and cured patients,length of hospital stay,hospitalization costs,rate of ICU admission,length of ICU stay,incidence of SIRS,and length of SIRS continue,Ranson scores,APACHE Ⅱ scores,computed tomographic severity index (CTSI) scores among the 3 groups were compared.Results One hundred and sixty-six patients with AP (119 males and 47 females) were included,and 76 were MAP,65 MSAP and 25 SAP.The average interval between AP onset and hospital admission was (2.27 ± 1.46) d.The number of improved and cured patients,length of hospital stay,hospitalization costs,rate of ICU admission,length of ICU stay,incidence of SIRS,and length of SIRS continue,Ranson scores,APACHE Ⅱ scores,CTSI scores increased with the severity of AP.The corresponding values in SAP group were 21 cases (84.0％),(23.8 ± 13.6) d,(53900 ± 30260) Yuan,48.0％ (12/25) and (5.76 ± 13.8) d,96.0％ (24/25) and (5.00 ± 2.40) d,(3.76 ± 1.30) score,(8.52 ± 4.24) score,(5.44 ± 3.48) score.Seventy-nine patients developed local complications,among them 34 was acute peripancreatic fluid collection,45 was acute necrosis collection.The incidence of acute necrosis collection in SAP group was significantly higher than that in MSAP group (68.0％ vs 44.6％,P =0.047),but the incidence of acute peripancreatic fluid collection in SAP group was significantly lower than that in MSAP group (16.0％ vs 46.2％,P =0.016).Organ failure occurred in 42 patients,among them 35 cases were respiratory failure,2 cases were renal failure,and 5 cases were respiratary and renal failure.The incidence of organ failure in SAP and MSAP group was 100％ and 26.2％,the difference between the two groups was statistically significant (P ＜ 0.05).Conclusions Classification of acute pancreatitis-2012 is a simple and convenient system,which can predict the severity of AP and appropriate for clinical application.

Objective To understand the malaria endemic characteristics and control measures in Caoxian County,Shan?dong Province,so as to summarize the experiences of malaria elimination. Methods The data of malaria endemic situation and control measures in Caoxian County from 1953 to 2014 were collected and descriptively analyzed,and the control effectiveness was evaluated. Results The incidence of malaria reduced from 13.25%in 1970 to 0.33%in 1983,and no malaria case was found in 1986. The goal of basic malaria elimination was achieved. The sporadic malaria infections were found from 2006 to 2010,and three imported malaria cases were found in Caoxian County from 2011 to 2014. Conclusion The effect of compre?hensive prevention and control measures taken in Caoxian County is significant,and the goal of malaria elimination has been reached. The imported malaria and secondary cases are future focuses of malaria control work.

[Objective]This study was designed to investigate the genetic evolution of the neuraminidase(NA)gene of seasonal A/H1N1 and 2009 novel A/H1N1 inflilenza virus,and discuss the genetic variation of influenza A virus.[Methods]The virus strains were separately isolated from the clinical samples collected in 2006 and 2009,and then identified as seasonal A/H1N1 and novel A/H1N1.The full length of the NA gene of these strains was amplified by RT-PCR.Then the genetic evolution and mutations of important functional sites were analyzed.[Results]The homology of NA gene between the 2009 novel A/H1N1 isolates and 2006 seasonal A/H1N1 isolates was low(77.9%～78.8%),so was the homology of NA gene between the 2009 novel A/H1N1 isolates and representative strains of different periods and 1979-2001 WHO recommended vaccine strains(78.1%～79.3%).But compared with the WHO recommended vaccine strains of 2009 novel A/H1N1,the homology reached more than 99%.The genetic evolution analysis revealed that NA gene of 2009 novel A/H1N1 had the closest genetic relationship with the swine influenza A virus(A/swine/Belgium/1/1983)from Eurasian Iineage,and some of the antigenic sites and neuraminidase active sites of NA gene of seasonal A/H1N1 were mutated after 2005.[Conclusion]The NA gene of 2009 novel A/H1N1 may originate from Eurasian Iineage of swine influenza virus.The variation of NA gene of seasonal A/H1N1 has occurred in a certain degree.Hence,it is very necessary to continuously monitor the variant of influenza A virus.

OBJECTIVETo explore the interaction between herbal medicines and western drugs based on CYP3A4 enzyme metabolism by using testotesrone as a probe in liver microsome metabolism system in vitro.

METHODThe mixed liver microsome enzymatic system consisting of rat liver microsomes by ultra-high-speed centrifuge was established. The substrate testosterone was added into the system and enzyme CYP3A4 metabolic activity was expressed by the output of 6beta-hydroxy-testosterone which was measured by HPLC method. The proper conditions for testotesrone metabolism in liver microsome system included substrate concentration, incubation time, pH and incubation temperature. When the conditions in vitro were determined, three kinds of Chinese herbal medicinal ingredients (Tetrahydropalmatine, neferine, panax notoginseng saponins) were diluted into different concentrations and incubated with testotesrone in the liver microsomes incubation system, respectively. The results were measured through metabolite production with or without the presence of Chinese medicines. We assessed the Chinese herbal medicinal ingredients effect on the metabolism of CYP3A4 enzyme through 6beta-hydroxy metabolite of testosterone production.

RESULTLiver microsomes were incubated in the system, the testosterone metabolited into 6beta-hydroxy testosterone. The metabolism conditions were proper at the concentration of testosterone 200 micromol x L(-1) which was incubated for 3.5 hours at 37 degrees C in pH 7.0, PBS 0.1 mol x L(-1). The inhibition of tetrahydropalmatine and panax notoginseng saponins on testotesrone were weak with IC50 > 100 micromol x L(-1). The neferine had a little inhibition on testotesrone metabolism, IC50 < 100 micromol L(-1).

CONCLUSIONTetrahydropalmatine and panax notoginseng saponins had no obvious effect on testotesrone metabolism. Neferine had a little effect on testotesrone metabolism. It prompted that drug-interaction could not be apparent between two kinds of Chinese medicines and the CYP3A4 enzyme substrate, Neferine could bring about drug-interaction.

Animals ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme System ; metabolism ; Drug Interactions ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Male ; Microsomes, Liver ; chemistry ; drug effects ; enzymology ; Rats ; Rats, Wistar ; Testosterone ; analysis ; pharmacokinetics

Perforated peptic ulcer is a common acute abdominal disease and requires emergency surgical treatment. Laparoscopic repair PPU was started early but progressed slowly. Compared with the popularization of minimally invasive concept of laparoscopic cholecystectomy, laparoscopic repair is still under the situation of disputation, low overall application rate and extremely unbalanced development in different regions and organizations. With the embodiment of minimally invasive advantages of laparoscopy and the technology progress, as well as the updating of surgeons′ ideas and the emergence of higher quality RCT studies, the proportion of PPU in laparoscopic treatment is gradually increasing. In order to make full use of the minimally invasive value and differential diagnostic value of laparoscopy, the laparoscopy-first approach can be adopted as a routine for appropriate PPU patients. The key to case selection and the physicians experience and proficiency. Laparoscopy should be actively adopted but should not be easily expanded. If necessary, the patients should be converted to open operation. If not the advantages of laparoscopy will be drowned.

Objective:To explore the mechanism of Honghua Xiaoyao Tablets in the treatment of premature ovarian failure (POF) using network pharmacology and molecular docking technology; To conduct further experimental verification.Methods:The active components and related targets of Honghua Xiaoyao Tablets were obtained using TCMSP and PubChem databases, and the related targets of POF were obtained by using GeneCards database. The Venn online tool was used to screen the intersection genes, and the STRING database was used to construct the PPI network. Then, GO function enrichment analysis and KEGG pathway enrichment analysis were performed on the intersecting genes through the Metescape database, and Cytoscape 3.7.1 software was used to construct the "active component-target" network and "Chinese materia medica-active component-target-key pathway-disease" network. Finally, molecular docking verification was carried out. Mice were divided into blank group, model group, and Honghua Xiaoyao Tablets group using a random number table method, with 8 mice in each group. Except for the blank group, mice in each group were treated with zona pellucida polypeptide 3 (ZP3) and Freund's Complete Adjuvant (FCA) to establish a mouse model of immune POF. Mice in the Honghua Xiaoyao Tablets group received Honghua Xiaoyao Tablets solution 0.56 g/kg for gavage, and the blank control group and the model group received saline for gavage for consecutive 4 weeks. The histopathological changes of the mouse ovary were observed by HE staining. Immunohistochemical staining was used to observe the expression of ESR1, and Western blot was used to detect the expressions of Akt and p-Akt.Results:A total of 80 intersection targets between Honghua Xiaoyao Tablets and POF were obtained, and the PPI network contained 44 core targets. The top 5 compounds in the topological analysis were formononetin, quercetin, Betulinic acid, Hydroxysafflor Yellow A and Baicalin, and the top 5 targets were PPARG, ESR1, AR, AKT1 and IL6. The molecular function of core genes was mainly receptor ligand activity, and its biological process mainly involved the positive regulation of cell migration. The cellular components mainly included membrane rafts, which were involved in signaling pathways such as cancer signaling pathway, proteoglycans in cancer, PI3K-Akt signaling pathway, and HIF-1 signaling pathway. "Chinese materia medica-component-target-pathway-disease" network showed that 8 kinds of Chinese materia medica in the Honghua Xiaoyao Tablets had important core components, among which Glycyrrhizae Radix et Rhizoma and Carthami Flos involved the most important core components. Molecular docking results showed that the active components had a good affinity with the core target. The experimental verification confirmed that Honghua Xiaoyao Tablets promoted follicular development, increased the expression of ESR1 in ovarian tissues and up-regulated the expression level of the key factor of Akt phosphorylation in the PI3K-Akt signaling pathway.Conclusions:The various active components of Honghua Xiaoyao Tablets may act on PPARG, ESR1 and other targets through multiple signaling pathways such as PI3K-Akt and HIF-1 to treat POF. The potential active components are mainly formononetin, quercetin, etc.

AIM:This study aimed to explore the induction of ferroptosis in non-small-cell lung cancer(NSCLC)cells by tubeimoside II(TBMS II)and to elucidate the underlying molecular mechanisms.METHODS:H460 NSCLC cells were cultured in vitro.Cell survival rates were assessed by using MTT assays,and doses of TBMS II resulting in below 50%survival were selected for further experimentation.Cell migration was evaluated using Transwell assays and the effects of TBMS II on H460 cell proliferation were assessed by colony formation assays.Flow cytometry and fluores-cence microscopy were used to assess changes in lipid peroxidation(lipid ROS),and the levels of GSH,T-AOC,MDA,and Fe2+were measured using commercial kits.Protein levels of GPX4,SLC7A11,FTH1,NCOA4,P62,and LC3 were examined using Western blot.Changes in mitochondrial structure were detected by transmission electron microscopy,and immunofluorescence was used to assess LC3 co-localization of FTH1 and NCOA4,as well as co-localization of LC3 and NCOA4 with lysosomes.RESULTS:Compared with the control group,TBMS II dose-dependently reduced H460 cell via-bility,migration,and clone formation,accompanied by the appearance of vacuoles within the cells.TBMS II treatment al-so led to decreased GSH and T-AOC levels,while increasing the cellular contents of MDA,indicating oxidative stress.Ad-ditionally,there was a decrease in the expression of the antioxidant proteins SLC7A11 and GPX4 in the cells,while lipid ROS and Fe2+levels were increased in proportion to the TBMS II concentration.The ferroptosis inhibitor ferrostatin-1 re-versed cell death caused by TBMS II,suggesting ferroptosis induction.Furthermore,increasing the TBMS II concentra-tion resulted in an upregulation of the autophagy marker proteins LC3 II/LC3 I and P62,indicative of increased autopha-gy.TBMS II also affected mitochondrial morphology in the cells,as seen in reduced mitochondrial fluorescence intensity.Protein expression of NCOA4 increased with higher TBMS II concentrations,while that of FTH1 decreased.Co-localiza-tion of LC3 II with FTH1 and NCOA4,as well as the lysosomal association of LC3 II and FTH1,also increased in a dose-dependent manner.CONCLUSION:TBMS II induces ferritinophagy in H460 cells,leading to decreased cell viability and increased ferroptosis.