ABSTRACT
@#Objective To optimize and verify the size exclusion chromatography-high performance liquid chromatography(SEC-HPLC) method for the determination of recombinant human growth hormone(rhGH)-Fc immunofusion protein polymer.Methods The multimer content of rhGH-Fc immunofusion protein was detected by SEC-HPLC.The detection conditions(salt concentration,mobile phase pH,flow rate,column temperature and column model) were optimized to observe the separation effect of the target proteins and polymers.The system suitability,specificity,linearity and range,precision,accuracy and limit of quantification of the method were verified.Results The optimized method was to use TSK-gel G2000SW_(x1)column(5 μm,7.8 mm × 300 mm),mobile phase of 50 mmol/L phosphate buffer(pH 6.80),detection wavelength of280 nm,injection volume of 100 μL,flow rate of 0.6 mL/min and column temperature of 45 ℃.The resolution of rhGHFc immunofusion protein and polymer,the theoretical plate number and the tailing factor all met the requirements;the peak time of rhGH-Fc immunofusion protein was the same as that of the control,while the peak time of GH national standard was different from that of the control,and the protein buffer showed no peak;the concentration of rhGH-Fc immunofusion protein was in the range of 0.307～1.842 mg/mL with good linear correlation between the peak area integral value and the injection volume(R~2=0.999 4);the RSD of peak area and purity in repeatability verification were 0.7% and 0.1%,respectively;the RSD of intermediate precision verification was 0.8%;the average recovery rate of accuracy verification was 99.1% with the RSD of 1.9%;the limit of quantification was 6 μg/mL.Conclusion The optimized SEC-HPLC method was used to detect the content of polymer in rhGH-Fc immunofusion protein with improved accuracy,and the column efficiency and separation were in accordance with the relevant requirements of Chinese Pharmacopoeia(Volume Ⅳ,2020edition),which could be used for the detection of polymer content in samples.
ABSTRACT
Objective To analyze the expression of SEC61G in oral squamous cell carcinoma (OSCC) tissues and cell lines and determine its correlations with the clinicopathological features and prognosis of patients with OSCC. Methods The expression of SEC61G in OSCC tissues and its diagnostic and prognostic value were detected in the TCGA database. The expression levels of SEC61G in paraffin-embedded OSCC tissues and adjacent normal tissue specimens of 64 patients with OSCC were detected by immunohistochemistry. The correlation of SEC61G expression in OSCC tissues with the clinicopathological features was analyzed. Kaplan-Meier survival curve analysis showed that the expression of SEC61G correlated with the overall survival time of patients with OSCC. Cox regression analysis was used to analyze prognostic influence factors. Results The expression of SEC61G in OSCC tissues was significantly higher than that in para-carcinoma tissues (P < 0.05), consistent with the results of the TCGA database analysis, and its expression was closely related to N stage and clinical stage (P < 0.05). The overall survival of patients with OSCC in the group with low SEC61G expression was significantly higher than that in the high SEC61G expression group (P < 0.05). N stage and SEC61G expression were the prognostic influence factors for patients with OSCC (P < 0.05). SEC61G expression was an independent predictor of the prognosis of patients with OSCC (P < 0.05, AUC=0.923). Conclusion SEC61G is highly expressed in OSCC tissues and associated with N stage and clinical stage. Its high expression is associated with poor prognosis of patients. It may be a diagnostic biomarker for OSCC.
ABSTRACT
@#Objective To develop and validate size exclusion column-high performance liquid chromatography(SEC-HPLC)for determination of the purity of bulk material of freeze-dried rabies vaccine for human use.Methods Chromatography column TSK-gel G6000PW_(XL)(7.8 mm × 30 cm,13 μm)was used for the determination(column temperature 30 ℃)with mobile phase of 0.1 mol/L PB buffer(pH 7.8)at a flow rate of 0.5 mL/min.The detection wavelength was 280 nm and injection volume was 20 μL.The method was validated for system suitability,specificity,precision and durability and determined for detection limit and quantitation limit,which was applied to analyze bulk material purity of freeze-dried rabies vaccine for human use of 3 batches of Vero cells and 1 batch of human diploid cells.Results The resolution of target protein spectrum peak of bulk material of reference sample and freeze-dried rabies vaccine for human use prepared with two substrates was more than 1.5 with a tailing factor less than 1.5;The blank solvent showed no absorption peak at the position of target protein peak with no interference in the determination;The RSDs of retention time and peak area in precision verification were both less than 2.0%;The quantitative limit was 10 μg/mL,and the detection limit was 4 μg/mL;The reference sample was injected three times continuously at three different detection wavelengths of 278,280 and 282 nm,and the RSDs of retention time and peak area were also less than 2.0%.The purity of 4 batches of freeze-dried rabies vaccine bulk material for human use was all more than 97%.Conclusion The developed SEC-HPLC for determination of the purity of freeze-dried rabies vaccine bulk material for human use showed good specificity,precision and durability,which provided a reliable method for the quality control of human rabies vaccine.
ABSTRACT
Objective: To investigate the clinical value of plasma scaffold protein SEC16A level and related models in the diagnosis of hepatitis B virus-related liver cirrhosis (HBV-LC) and hepatocellular carcinoma (HBV-HCC). Methods: Patients with HBV-LC and HBV-HCC and a healthy control group diagnosed by clinical, laboratory examination, imaging, and liver histopathology at the Third Hospital of Hebei Medical University between June 2017 and October 2021 were selected. Plasma SEC16A level was detected using an enzyme-linked immunosorbent assay (ELISA). Serum alpha-fetoprotein (AFP) was detected using an electrochemiluminescence instrument. SPSS 26.0 and MedCalc 15.0 statistical software were used to analyze the relationship between plasma SEC16A levels and the occurrence and development of liver cirrhosis and liver cancer. A sequential logistic regression model was used to analyze relevant factors. SEC16A was established through a joint diagnostic model. Receiver operating characteristic curve was used to evaluate the clinical efficacy of the model for liver cirrhosis and hepatocellular carcinoma diagnosis. Pearson correlation analysis was used to identify the influencing factors of novel diagnostic biomarkers. Results: A total of 60 cases of healthy controls, 60 cases of HBV-LC, and 52 cases of HBV-HCC were included. The average levels of plasma SEC16A were (7.41 ± 1.66) ng/ml, (10.26 ± 1.86) ng/ml, (12.79 ± 1.49) ng /ml, respectively, with P < 0.001. The sensitivity and specificity of SEC16A in the diagnosis of liver cirrhosis and hepatocellular carcinoma were 69.44% and 71.05%, and 89.36% and 88.89%, respectively. SEC16A, age, and AFP were independent risk factors for the occurrence of HBV-LC and HCC. SAA diagnostic cut-off values, sensitivity, and specificity were 26.21 and 31.46, 77.78% and 81.58%, and 87.23% and 97.22%, respectively. The sensitivity and specificity for HBV-HCC early diagnosis were 80.95% and 97.22%, respectively. Pearson correlation analysis showed that AFP level was positively correlated with alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), and γ-glutamyltransferase (GGT) with P < 0.01, while the serum SEC16A level was only slightly positively correlated with ALT and AST in the liver cirrhosis group (r = 0.268 and 0.260, respectively, P < 0.05). Conclusion: Plasma SEC16A can be used as a diagnostic marker for hepatitis B-related liver cirrhosis and hepatocellular carcinoma. SEC16A, combined with age and the AFP diagnostic model with SAA, can significantly improve the rate of HBV-LC and HBV-HCC early diagnosis. Additionally, its application is helpful for the diagnosis and differential diagnosis of the progression of HBV-related diseases.
Subject(s)
Humans , Carcinoma, Hepatocellular/pathology , Liver Neoplasms/pathology , alpha-Fetoproteins/metabolism , Endoplasmic Reticulum/metabolism , Golgi Apparatus/metabolism , Vesicular Transport Proteins , Liver Cirrhosis/complications , Hepatitis B/complications , ROC Curve , Hepatitis B virus/metabolism , Biomarkers, TumorABSTRACT
The general secretory (Sec) pathway represents a common mechanism by which bacteria secrete proteins, including virulence factors, into the extracytoplasmic milieu. However, there is little information about this system, as well as its associated secretory proteins, in relation to the fire blight pathogen Erwinia amylovora. In this study, data mining revealed that E. amylovora harbors all of the essential components of the Sec system. Based on this information, we identified putative Sec-dependent secretory proteases in E. amylovora on a genome-wide scale. Using the programs SignalP, LipoP, and Phobius, a total of 15 putative proteases were predicted to contain the N-terminal signal peptides (SPs) that might link them to the Sec-dependent pathway. The activities of the predicted SPs were further validated using an Escherichia coli-based alkaline phosphatase (PhoA) gene fusion system that confirmed their extracytoplasmic property. Transcriptional analyses showed that the expression of 11 of the 15 extracytoplasmic protease genes increased significantly when E. amylovora was used to inoculate immature pears, suggesting their potential roles in plant infection. The results of this study support the suggestion that E. amylovora might employ the Sec system to secrete a suite of proteases to enable successful infection of plants, and shed new light on the interaction of E. amylovora with host plants.
Subject(s)
Erwinia amylovora/metabolism , Escherichia coli/genetics , Peptide Hydrolases/genetics , Plant Diseases/microbiology , Pyrus/microbiologyABSTRACT
The general secretory (Sec) pathway represents a common mechanism by which bacteria secrete proteins, including virulence factors, into the extracytoplasmic milieu. However, there is little information about this system, as well as its associated secretory proteins, in relation to the fire blight pathogen Erwinia amylovora. In this study, data mining revealed that E. amylovora harbors all of the essential components of the Sec system. Based on this information, we identified putative Sec-dependent secretory proteases in E. amylovora on a genome-wide scale. Using the programs SignalP, LipoP, and Phobius, a total of 15 putative proteases were predicted to contain the N-terminal signal peptides (SPs) that might link them to the Sec-dependent pathway. The activities of the predicted SPs were further validated using an Escherichia coli-based alkaline phosphatase (PhoA) gene fusion system that confirmed their extracytoplasmic property. Transcriptional analyses showed that the expression of 11 of the 15 extracytoplasmic protease genes increased significantly when E. amylovora was used to inoculate immature pears, suggesting their potential roles in plant infection. The results of this study support the suggestion that E. amylovora might employ the Sec system to secrete a suite of proteases to enable successful infection of plants, and shed new light on the interaction of E. amylovora with host plants.
ABSTRACT
Objective: A new table of standards of commodity classes of Saposhnikovia Radix is formed through literature market and local area survey. To explore the scientific nature and practicability of new standards of commodity classes of Saposhnikovia Radix, according to the appearance and index components related to quality of Saposhnikovia Radix, through statistical analysis and comparison of characteristic spectrum. Methods: Samples of different specifications and grades were collected from Northeast China, Inner Mongolia, Hebei, and the other main production areas, mainstream medicine market. Measuring samples appearance, extractum, characteristic spectrum of four chromones and analyzing data and discussing the relationship between standards of commodity classes and the appearance and index components to provide datas and theoretical support for the new table. Results: According to appearance character, the standards of commodity classes of Saposhnikovia Radix divided into three specifications of wild, imitates wild, cultivated were science based. At the same time, it is suggested to increase the content of the cimifugin as the index to distinguish between wild and cultivated Saposhnikovia Radix. Conclusion: There were significant differences in appearance and index components between different standards of Saposhnikovia Radix. Whether different classes of Saposhnikovia Radix only depend on size remains to be discussed.
ABSTRACT
Bovine mastitis has a negative impact on milk production and can pose risks to public health. The present study aimed to evaluate the quality of bovine milk from small farms in the Botucatu/SP region. Somatic cell counts (SCC), identification of pathogens involved in mastitis, and sensitivity antimicrobial profile of staphylococci isolated were performed. The presence of enterotoxin encoding genes in isolates of staphylococci obtained from milk was investigated. Milk samples from individual mammary quarters of cows were submitted to the California mastitis test (CMT) and SCC. Of the 239 dairy cows from 21 dairy herds evaluated (mean = 11.4 animals/property), two cows (0.8%) presented clinical mastitis and 86 (35.9%) subclinical mastitis. Bacterial culture was performed in 177 quarter milk samples. Staphylococci were identified in 55 (31.1%), corynebacteria in 45 (25.4%), streptococci in 25 (14.1%) and coliforms in four (2.3%) milk samples. Average SCC from culture-positive samples was 1598x103 cells/mL, in case of staphylococci was 1362x103 cells/ml, streptococci was 2857x103 cells/mL, corynebacteria was 976x103 cells/mL and in the cases of coliforms 1161x103 cells/mL were obtained. Staphylococci showed a high sensitivity (>95%) to cephalothin, cotrimoxazole, enrofloxacin, and gentamicin, with a 41.2% resistance to penicillin and 11.8% to oxacillin. Both coagulase positive (CPS) and negative staphylococci (CNS) carried genes encoding enterotoxins in 21.6% of the first group and 41.9% in the second. The sea gene was the most detected 45.8% (n=24) between them, followed by seb with 29.2% and sec with 25.0%. The sed gene was not identified. We highlight the potential risk to public health in the possibility of strains of Staphylococcus spp. enterotoxin-producing genes that can cause staphylococcal food poisoning.(AU)
A mastite bovina impacta negativamente a produção leiteira e pode acarretar riscos à saúde pública. O presente estudo teve como objetivo a avaliação da qualidade do leite bovino proveniente de pequenas propriedades na região de Botucatu/SP. Foi realizada a contagem de células somáticas (CCS), identificação dos patógenos envolvidos nas mastites, e realizado o perfil de sensibilidade aos antimicrobianos dos estafilococos isolados. Pesquisou-se a presença de genes codificadores de enterotoxinas em isolados de estafilococos obtidos a partir do leite mastítico. Amostras de leite de quartos mamários individuais de vacas foram submetidas ao "California mastitis test" (CMT) e à CCS. Das 239 vacas em lactação provenientes de 21 rebanhos leiteiros avaliados (média = 11,4 animais/propriedade), dois (0,8%) animais apresentaram mastite clínica e, 86 (35,9%) mastite subclínica. 177 amostras de leite foram cultivadas em ágar sangue bovino 5% e ágar MacConckey e obteve-se 55 (31,1%) Staphylococcus spp., 25 (14,1%) Streptococcus spp., 45 (25,4%) Corynebacterium spp. e quatro (2,3%) coliformes. A média da CCS das amostras procedentes de todos os quartos mamários infectados avaliados foi de 1598x103 células/mL, enquanto que nos casos que foram isolados Staphylococcus spp. foi de 1362x103 células/mL, Streptococcus spp. 2857x103 células/mL, Corynebacterium spp. de 976x103 células/mL e nos casos de coliformes 1161x103 células/mL. Os estafilococos revelaram grande sensibilidade (>95%) à cefalotina, cotrimoxazol, enrofloxacina e gentamicina, com resistência de 41,2% à penicilina e 11,8% à oxacilina. Tanto estafilococos coagulase positivos (ECP) como negativos (ECN) revelaram genes codificadores de enterotoxinas em 21,6% do primeiro grupo e 41,9% no segundo. O gene sea foi o mais detectado 45,8% (n=24), seguido pelo seb com 29,2% e sec com 25,0%. O gene codificador da sed não foi identificado. Frente aos resultados, destaca-se o risco potencial à saúde pública pela possibilidade de veiculação de linhagens de Staphylococcus spp. carreadores de genes produtores de enterotoxinas, podendo ocasionar toxi-infecções alimentares.(AU)
Subject(s)
Staphylococcus/isolation & purification , Cell Count/veterinary , Milk/microbiology , Mastitis, Bovine/diagnosis , Cattle/microbiology , DairyingABSTRACT
Objective: To enrich the gene mutation sites and accumulate treatment experience of congenital dyserythropoietic anemia (CDA) type Ⅱ by reporting one case of CDA patient with new mutation site of SEC23B and was successfully treated by homozygous allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: The mutation within SEC23B gene in a child case with the reduced hemoglobin for more than 3 months, and his family were analyzed in combination with literatures review. Results: A 3-day 5-month female child was admitted due to "decreasing hemoglobin for more than 3 months" , blood routine test showed HGB 44 g/L, positive for acid hemolysis test (Ham test) . Bone marrow showed that the proportion of erythroid line was 69%, mainly middle and late juvenile erythrocytes, binuclear and odd nucleated erythrocytes could be observed, and nuclear fragmentation and nuclear budding could be seen occasionally in nucleated erythrocytes, transmission electron microscopy disclosed that bone marrow harbored the typical double-layer membrane structure of nuclear erythrocytes. There were two unreported new mutation sites in the SEC23B gene, including 1504 G>C/wt and c. 2254-2255 insert A/wt. The two mutations were derived from the father and mother of the child respectively. At the late stage, the child was successfully treated with allo-HSCT, the original mutation turned negative. Conclusion: This study reported the mutation type of SEC23B gene insertion for the first time in China. Allo-HSCT could be utilized as a treatment for CDA.
Subject(s)
Female , Humans , Anemia, Dyserythropoietic, Congenital/genetics , China , Erythroblasts , Mutation , Vesicular Transport Proteins/geneticsABSTRACT
Objective@#To enrich the gene mutation sites and accumulate treatment experience of congenital dyserythropoietic anemia (CDA) type Ⅱ by reporting one case of CDA patient with new mutation site of SEC23B and was successfully treated by homozygous allogeneic hematopoietic stem cell transplantation (allo-HSCT) .@*Methods@#The mutation within SEC23B gene in a child case with the reduced hemoglobin for more than 3 months, and his family were analyzed in combination with literatures review.@*Results@#A 3-day 5-month female child was admitted due to "decreasing hemoglobin for more than 3 months" , blood routine test showed HGB 44 g/L, positive for acid hemolysis test (Ham test) . Bone marrow showed that the proportion of erythroid line was 69%, mainly middle and late juvenile erythrocytes, binuclear and odd nucleated erythrocytes could be observed, and nuclear fragmentation and nuclear budding could be seen occasionally in nucleated erythrocytes, transmission electron microscopy disclosed that bone marrow harbored the typical double-layer membrane structure of nuclear erythrocytes. There were two unreported new mutation sites in the SEC23B gene, including 1504 G>C/wt and c. 2254-2255 insert A/wt. The two mutations were derived from the father and mother of the child respectively. At the late stage, the child was successfully treated with allo-HSCT, the original mutation turned negative.@*Conclusion@#This study reported the mutation type of SEC23B gene insertion for the first time in China. Allo-HSCT could be utilized as a treatment for CDA.
ABSTRACT
The exocyst is a highly conserved eight-subunit protein complex (EXOC1–8) involved in the targeting and docking of exocytic vesicles translocating from the trans-Golgi network to various sites in renal cells. EXOC5 is a central exocyst component because it connects EXOC6, bound to the vesicles exiting the trans-Golgi network via the small GTPase RAB8, to the rest of the exocyst complex at the plasma membrane. In the kidney, the exocyst complex is involved in primary ciliognesis, cystogenesis, and tubulogenesis. The exocyst, and its regulators, have also been found in urinary extracellular vesicles, and may be centrally involved in urocrine signaling and repair following acute kidney injury. The exocyst is centrally involved in the development of other organs, including the eye, ear, and heart. The exocyst is regulated by many different small GTPases of the RHO, RAL, RAB, and ARF families. The small GTPases, and their guanine nucleotide exchange factors and GTPase-activating proteins, likely give the exocyst specificity of function. The recent development of a floxed Exoc5 mouse line will aid researchers in studying the role of the exocyst in multiple cells and organ types by allowing for tissue-specific knockout, in conjunction with Cre-driver mouse lines.
Subject(s)
Animals , Humans , Mice , Acute Kidney Injury , Cell Membrane , Ear , Exocytosis , Extracellular Vesicles , GTP Phosphohydrolases , GTPase-Activating Proteins , Guanine Nucleotide Exchange Factors , Heart , Kidney , Monomeric GTP-Binding Proteins , Sensitivity and Specificity , trans-Golgi NetworkABSTRACT
BACKGROUND: This study was performed in order to examine the effect of intrathecal sec-O-glucosylhamaudol (SOG), an extract from the root of the Peucedanum japonicum Thunb., on incisional pain in a rat model. METHODS: The intrathecal catheter was inserted in male Sprague-Dawley rats (n = 55). The postoperative pain model was made and paw withdrawal thresholds (PWTs) were evaluated. Rats were randomly treated with a vehicle (70% dimethyl sulfoxide) and SOG (10 μg, 30 μg, 100 μg, and 300 μg) intrathecally, and PWT was observed for four hours. Dose-responsiveness and ED50 values were calculated. Naloxone was administered 10 min prior to treatment of SOG 300 μg in order to assess the involvement of SOG with an opioid receptor. The protein levels of the δ-opioid receptor, κ-opioid receptor, and μ-opioid receptor (MOR) were analyzed by Western blotting of the spinal cord. RESULTS: Intrathecal SOG significantly increased PWT in a dose-dependent manner. Maximum effects were achieved at a dose of 300 μg at 60 min after SOG administration, and the maximal possible effect was 85.35% at that time. The medial effective dose of intrathecal SOG was 191.3 μg (95% confidence interval, 102.3–357.8). The antinociceptive effects of SOG (300 μg) were significantly reverted until 60 min by naloxone. The protein levels of MOR were decreased by administration of SOG. CONCLUSIONS: Intrathecal SOG showed a significant antinociceptive effect on the postoperative pain model and reverted by naloxone. The expression of MOR were changed by SOG. The effects of SOG seem to involve the MOR.
Subject(s)
Animals , Humans , Male , Rats , Analgesia , Blotting, Western , Catheters , Dimethyl Sulfoxide , Hyperalgesia , Models, Animal , Naloxone , Nociceptive Pain , Pain, Postoperative , Rats, Sprague-Dawley , Receptors, Opioid , Spinal CordABSTRACT
Hemorrhage often occurs during caesarean section,which can lead to severe postpartum hemorrhage if not treated in time.Effective treatment can avoid serious bleeding and following complications.This paper discussed the advantages and disadvantages of various hemostasis schemes,which included uterotonics,uterine cavity tamponade,uterine compression suture,and vascular occlusion.Any hemostasis selected should be used as early as possible to achieve sufficient effect and reduce the morbidity of complications after massive hemorrhage.
ABSTRACT
Background: The inherited disorders of haemoglobin are the commonest single gene disorder of the world population. Sickle cell anaemia being the most widespread and numerically the most important haemoglobinopathy in the world today. The major features of sickle cell disease (SCD) in most patients are life-long anaemia and the consequences of recurrent vaso-occlusion. Many complications of SCD involve anaemia, vasoocclusion etc. Impairment of pulmonary function is a common complication of SCD. The patients suffering from this disease frequently present with complaints referable to the pulmonary system although other systems are also involved. Hence present study was carried out to determine the difference in the pulmonary function tests between male and female sickle cell disease patients ( HbSS ),sickle cell trait patients(HbAS)and compare with normal controls ( HbAA) non-sicklers. Aim and Objectives: 1) To study the pulmonary alterations in cases of homozygous SS & heterozygous AS Sickle Cell Disorder subjects.2) To compare the parameters with normal healthy controls of both sexes & assess the importance of PFT in "Steady State"(free from complications or crisis)as an objective evidence to predict the risk of "Sickle Cell Chronic Lung Disease'' in future. Method:Across sectional study was done in 50 cases of SCD (22HbSS& 28HbAS) and age and sex matched normal 50 HbAAcontrols. From the various measured pulmonary function test( PFT )parameters Forced Vital Capacity( FVC),Forced Expiratory Volume in 1 sec(FEV1) Peak Expiratory Flow Rate(PEFR)in both sexes were selected for the study. The data collected was subjected to statistical analysis involving computation of Mean, Standard deviation, Independent Ttest. Result: Mean value of FVC, FEV1 and PEFR were found to be significantly lower than normal controls, but the difference in FVC between HbAS and HbSS turned out to be statistically non-significant whereas the difference in FEV1, PEFR between HbAS and HbSS was statistically significant.The lung function indices were lower in females than males of the sickle cell patients. Conclusion: There were significant reductions in pulmonary function test parameters in sickle cell anaemia patients as compared to normal controls of a similar age and gender.It is indicative of mixed pattern (both restrictive and obstructive) lung impairment in sickle cell anaemia.
ABSTRACT
Aim To explore the antidepressant effect and the mechanism of self-made prescription of strengthening spleen and replenishing Qi ( SMP-SS-RQ) . Methods The depression model of chronic un-predictable mild stress ( CUMS ) was established in mice. The animals were randomly divided into control group, model group, SMP-SSRQ low, middle and high dose groups (8, 16, 32 mg·kg-1), with 12 mice for each group. The control group and the model group were given the same volume of saline, and other groups were given corresponding dose of SMP-SSRQ. Animals of each group were administered by gavage twice a day for two weeks. Behavioral indexes of mice were deter-mined by open field experiment, sugar consumption test, forced swimming test and tail suspension test. Quantitative real-time PCR (qPCR) and Western blot were respectively performed to detect mRNA and pro-tein expression levels of Igsf11 , Pdia2 and Sec14 l2 inprefrontal cortex and hippocampus. Results Com-pared with model group, depressed mice' s horizontal mobile score, upright number and the sugar water pref-erence index increased and FST as well as TST de-creased in all SMP-SSRQ groups(P<0.05). The de-pression symptoms in mice were obviously improved by SMP-SSRQ therapy. Low and high doses of SMP-SSRQ significantly reduced mRNA and protein expression lev-el of Igsf11 , Pdia2 and Sec14 l2 in the prefrontal cortex and hippocampus of the depressed mice, presenting significant statistical difference compared with model group ( P<0.05 ) . Conclusions SMP-SSRQ can ef-fectively improve mouse depressive behavior, and its mechanism may be related to the down-regulation of Igsf11 , Pdia2 and Sec14 l2 in mouse prefrontal cortex and hippocampus.
ABSTRACT
Sec61β, a subunit of the Sec61 translocon complex, is not essential in yeast and commonly used as a marker of endoplasmic reticulum (ER). In higher eukaryotes, such as Drosophila, deletion of Sec61β causes lethality, but its physiological role is unclear. Here, we show that Sec61β interacts directly with microtubules. Overexpression of Sec61β containing small epitope tags, but not a RFP tag, induces dramatic bundling of the ER and microtubule. A basic region in the cytosolic domain of Sec61β is critical for microtubule association. Depletion of Sec61β induces ER stress in both mammalian cells and Caenorhabditis elegans, and subsequent restoration of ER homeostasis correlates with the microtubule binding ability of Sec61β. Loss of Sec61β causes increased mobility of translocon complexes and reduced level of membrane-bound ribosomes. These results suggest that Sec61β may stabilize protein translocation by linking translocon complex to microtubule and provide insight into the physiological function of ER-microtubule interaction.
Subject(s)
Animals , Humans , COS Cells , Caenorhabditis elegans Proteins , Genetics , Metabolism , Cell Line, Tumor , Chlorocebus aethiops , Endoplasmic Reticulum , Metabolism , Homeostasis , Microtubules , Metabolism , SEC Translocation Channels , Genetics , MetabolismABSTRACT
Resumen:La Entomología Forense es una ciencia que implica el estudio de las especies antropofágicas que practican un ciclo vital en el cadáver. Contribuye en la determinación del Intervalo posmortem. Los insectos de importancia forense que se estudiaron pertenecen a la orden Dípteros y Coleópteros. En el estudio realizado en el municipio de Pucarani, ubicado en la provinciaLos Andes del Departamento La Paz a 3852 m.s.n.m. entre otoño e invierno, se obtuvo dos ejemplares de cerdo S. scrofa, en dos ambientes: abierto (cerdo A) y cerrado: (cerdo B). Se identificó 339 especímenes entre dípteros y coleópteros. No se encontraron diferencias significativas en la sucesión de entomofauna entre ambos ambientes, sin embargo predominó la familia Fannide sp. en el lugar cerrado por ser un ambiente húmedo. Los dípteros de mayor importancia son: Sarconesia Chlorogaster, Clorobrachycoma splendida, que pertenecen a la familia Calliphoridae, que se mantuvo en constante actividaden su estadio larval y su forma adulta en los cuatro estados de descomposición a temperaturas de 14 - 16°C, humedad relativa de 30%; no se encontró a Calliphora nigribasis en la descomposición avanzada; Sarconesiopsis magellánica solo se presenta en la etapa Fresco. Entre los Coleópteros: Oxelitrum apicale apareció desde la etapa hinchado. La sucesión de entomofauna cadavérica y su particularidad en cada etapa de descomposición, es el elemento que nos ayudaría a determinar el IPM, que dependerá de las condiciones climáticas a gran altitud como Bolivia.
Abstract:Forensic entomology is a science that involves the study of the cannibalistic species that plays a vital cycle in the body. It helps to determinate the postmortem interval. Insects with forensic importance that were studied belong to the order Diptera and Coleoptera. In the study conducted in the Municipality of Pucarani, located in the province of Los Andes La Paz Department at 3852 meters between autumn and winter, two copies of pork S. scrofa was obtained in two environments: open (pork A), closed (pig B). 339 specimens between Diptera and Coleoptera were identified. No significant differences were found in the entomofauna sequence between the two environments, however the Fannide sp. In the place closed for being a humid environment. The most important dipterans are: Sarconesia Chlorogaster, Clorobrachycoma splendida, belonging to the family Calliphoridae, which remained in constant activity in its larval stage and its adult form in the four decaying states at temperatures of 14 - 16 ° C, humidity Relative to 30%; Calliphora nigribasis was not found in advanced decomposition; Magellanic sarconesiopsis only occurs in the Fresco stage. Among the Coleoptera: Oxelitrum apicale appeared from the swollen stage. The succession of cadaveric entomofauna and its particularity in each stage of decomposition is the element that would help us determine the MPI, which will depend on high altitude climatic conditions like Bolivia.
Subject(s)
Animals , Swine , Coleoptera , Bolivia , Cadaver , Forensic Sciences , Diptera , Entomology , InsectaABSTRACT
Objective To understand the situation of college students' awareness about innovative credit, analyze the influencing factors, in order to provide a scientific basis for constructing innovation credits and promoting the double strategy. Method The undergraduates of a medical university who were still at school in April 2015 were selected, and by using a stratified cluster sampling method, 450 students were chosen to be conducted by self-administered questionnaire survey to understand the cognitive status of in-novation of credit system in college students. Chi square test was used to compare the passing rates of students in different demographic characteristics, and the relationship between scores was analyzed by linear regression analysis. Result A total of 439 valid questionnaires were collected , and the total scores of innovative knowledge, attitudes and behaviors of 439 undergraduates were 15.95％ (70/439), 36.44％ (160/439) and 32.80％(144/439) respectively. In the part of knowledge innovation credits, girls' passing rate was higher than boys ( χ2=4.010,P=0.045. In the part of attitude, high grade group students had more positive attitude than the low grade group of students ( χ2=6.227,P=0.0013). In the part of behavior; higher grade students with innovative credits had higher pass rates than those in lower grades (chi, 2=7.781, P=0.005), and boys had a higher rate of passing than girls ( χ2=6.658,P=0.010). The total score of knowledge was positively linear with the total score of attitude and behavior. Conclusion College Students' awareness of innovation credit rate is low, but the innovative attitude and behavior is positive. The higher the awareness rate is conducive to the cultivation of attitude and behavior, so it is necessary in medical colleges to carry out innovation of credit and promote the cultivation of innovative consciousness.
ABSTRACT
BACKGROUND: The root of Peucedanum japonicum Thunb., a perennial herb found in Japan, the Philippines, China, and Korea, is used as an analgesic. In a previous study, sec-O-glucosylhamaudol (SOG) showed an analgesic effect. This study was performed to examine the antinociceptive effect of intrathecal SOG in the formalin test. METHODS: Male Sprague-Dawley rats were implanted with an intrathecal catheter. Rats were randomly treated with a vehicle and SOG (10 µg, 30 µg, 60 µg, and 100 µg) before formalin injection. Five percent formalin was injected into the hind-paw, and a biphasic reaction followed, consisting of flinching and licking behaviors (phase 1, 0–10 min; phase 2, 10–60 min). Naloxone was injected 10 min before administration of SOG 100 µg to evaluate the involvement of SOG with an opioid receptor. Dose-responsiveness and ED50 values were calculated. RESULTS: Intrathecal SOG showed a significant reduction of the flinching responses at both phases in a dose-dependent manner. Significant effects were showed from the dose of 30 µg and maximum effects were achieved at a dose of 100 µg in both phases. The ED50 value (95% confidence intervals) of intrathecal SOG was 30.3 (25.8–35.5) µg during phase 1, and 48.0 (41.4–55.7) during phase 2. The antinociceptive effects of SOG (100 µg) were significantly reverted at both phases of the formalin test by naloxone. CONCLUSIONS: These results demonstrate that intrathecal SOG has a very strong antinociceptive effect in the formalin test and it seems the effect is related to an opioid receptor.
Subject(s)
Animals , Humans , Male , Rats , Analgesia , Catheters , China , Formaldehyde , Japan , Korea , Naloxone , Nociception , Pain Measurement , Philippines , Rats, Sprague-Dawley , Receptors, OpioidABSTRACT
Objective To systematically evaluate the associations between SEC16B polymorphisms and body mass index (BMI) or risk of obesity in different ethnic populations.Methods A literature retrieval was carried out by using Wanfangdata,Chinese National Knowledge Infrastructure (CNKI),China Science and Technology Journal Database (VIP databases),PubMed,Embase,Web of Science,NIH GWAS catalog databases to collect the research papers published between 2009 and 2015 on the associations between SEC16B polymorphisms and BMI or risk of obesity.Summary beta estimates (βs),odds ratios (ORs) and 95％ confidence intervals (CIs) were calculated to assess the strength for the BMI analyses and obesity status.Meta-analysis was performed with Stata 12.0 software.Results Totally 15 papers for rs10913469 and 13 papers for rs543874 were included in this Meta-analysis.Under additive genetic model,rs10913469 and rs543874 in SEC16B gene were positively associated with BMI,and the combined β was 0.04 (95％CI:0.03-0.05) and 0.03 (95％CI:0.02-0.04),respectively,and rs10913469 and rs543874 were also associated with the risk of obesity,and the combined OR was 1.11 (95％CI:1.08-1.15) and 1.28 (95％CI:1.20-1.36),respectively.There were no significant differences among subgroups of ethnicity,different age groups and literatures with different quality.Conclusion rs10913469 and rs543874 in SEC16B gene are significantly associated with BMI and the risk of obesity,and C allele ofrs10913469 and G allele ofrs543874 increase the risk for obesity in different ethnic populations.