Mechanism of action and exogenous supplementation of vitamin D in Crohn's disease / 中国当代儿科杂志

Vitamin D can not only regulate calcium and phosphorus metabolism, but also exert an immunoregulatory effect. Vitamin D deficiency is common in patients with Crohn's disease (CD). Studies have shown that vitamin D is associated with CD and other autoimmune diseases and can improve the condition of patients with CD and promote their recovery by regulating intestinal immunity, repairing the intestinal mucosal barrier, inhibiting intestinal fibrosis, enhancing the response to infliximab, and regulating intestinal microbiota. Exogenous vitamin D supplementation can induce disease remission while increasing the serum level of vitamin D. However, only a few randomized, double-blind, and placebo-controlled trials have investigated the therapeutic effect of vitamin D in CD, and the optimal form of vitamin D supplementation, the specific dosage of vitamin D supplementation, and the optimal serum maintenance concentration of vitamin D remain to be clarified. This article mainly discusses the mechanism of action of vitamin D in CD and the beneficial effect of exogenous vitamin D supplementation on CD.

Retrospective Analysis of Death Cases of Oral Diphenidol Hydrochloride Poisoning / 法医学杂志

OBJECTIVES@#To explore the characteristics of postmortem examination, chemical examination and scene investigation of deaths caused by oral diphenidol hydrochloride poisoning, and so as to provide a reference for proper settlement and prevention of such deaths.@*METHODS@#The data of 22 deaths caused by oral diphenidol hydrochloride poisoning in a city from January 2018 to August 2020 were collected, including case details, scene investigations, autopsies, chemical examinations and digital evidence. Thirty-one cases of deaths caused by oral diphenidol hydrochloride poisoning reported in previous literature were also collected.@*RESULTS@#In the 53 oral diphenidol hydrochloride poisoning death cases, 50 cases were suicide, 2 cases were accidental, while 1 case was undetermined. Fifty-two cases were found in the medical records or crime scene investigation reports with doses ranging from 775 mg to 12 500 mg, and 23 deceased were detected with postmortem blood concentrations ranging from 2.71 mg/L to 83.1 mg/L. Clinical symptoms were recorded in 6 patients, including conscious disturbance and convulsion. Among the 45 cases which were performed with external examination, 23 cases autopsied.@*CONCLUSIONS@#Most of the deceased of oral diphenidol hydrochloride poisoning were suicide. No significant correlation was found between dose and blood concentration through the retrospective analysis of cases.

GPRC5B protects osteoarthritis by regulation of autophagy signaling

Osteoarthritis (OA) is one of the most common chronic diseases in the world. However, current treatment modalities mainly relieve pain and inhibit cartilage degradation, but do not promote cartilage regeneration. In this study, we show that G protein-coupled receptor class C group 5 member B (GPRC5B), an orphan G-protein-couple receptor, not only inhibits cartilage degradation, but also increases cartilage regeneration and thereby is protective against OA. We observed that Gprc5b deficient chondrocytes had an upregulation of cartilage catabolic gene expression, along with downregulation of anabolic genes in vitro. Furthermore, mice deficient in Gprc5b displayed a more severe OA phenotype in the destabilization of the medial meniscus (DMM) induced OA mouse model, with upregulation of cartilage catabolic factors and downregulation of anabolic factors, consistent with our in vitro findings. Overexpression of Gprc5b by lentiviral vectors alleviated the cartilage degeneration in DMM-induced OA mouse model by inhibiting cartilage degradation and promoting regeneration. We also assessed the molecular mechanisms downstream of Gprc5b that may mediate these observed effects and identify the role of protein kinase B (AKT)-mammalian target of rapamycin (mTOR)-autophagy signaling pathway. Thus, we demonstrate an integral role of GPRC5B in OA pathogenesis, and activation of GPRC5B has the potential in preventing the progression of OA.

Mechanism of Buyang Huanwutang in Regulating Macrophage Cell Polarization Based on TLR4/NF-κB/NLRP3 Pathway / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Buyang Huanwutang in regulating macrophage polarization based on the Toll-like receptor 4 （TLR4） / nuclear factor-κB （NF-κB） / nucleotide-binding oligomerization domain-like receptor 3 （NLRP3） pathway. MethodRAW264.7 macrophages were intervened with lipopolysaccharide （LPS） of different concentrations （0， 1.25， 2.5， 5， 10， 20， 40， and 80 mg·L-1） for 24 hours. Cell Counting Kit-8 （CCK-8） assay was used to determine the cell viability of RAW264.7 macrophages. The optimal concentration was chosen to establish an in vitro inflammation model induced by LPS. Cells were divided into a blank group （20% blank serum）， a model group （20% blank serum + 10 mg·L-1 LPS）， a model control group （20% FBS + 10 mg·L-1 LPS）， low-， medium-， and high-dose （5%， 10%， and 20%） Buyang Huanwutang-containing serum groups， a high-dose （20%） Buyang Huanwutang combined with NLRP3 inhibitor MCC950 （50 μmol·L-1） group， a high-dose （20%） Buyang Huanwutang combined with reactive oxygen species （ROS） inhibitor NAC （10 μmol·L-1） group， and a high-dose （20%） Buyang Huanwutang combined with NF-κB inhibitor PDTC （10 μmol·L-1） group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the expression of interleukin-1β （IL-1β）， interleukin-18 （IL-18）， and tumor necrosis factor-α （TNF-α） in RAW264.7 macrophages. Flow cytometry was employed to measure ROS levels in macrophages. Western blot was used to determine the protein expression of M1-type macrophage-related factors inducible nitric oxide synthase （iNOS） and TNF-α， M2-type macrophage-related factors arginase-1 （Arg-1） and interleukin-10 （IL-10）， as well as the proteins in the TLR4/NF-κB/NLRP3 pathway. ResultCCK-8 results indicated that under 10 mg·L-1 LPS stimulation， RAW264.7 macrophages exhibited the highest cell viability （P<0.01）. Compared with the blank group， the model group showed significantly increased levels of IL-1β， IL-18， and TNF-α （P<0.05，P<0.01）， increased ROS expression （P<0.05，P<0.01）， increased protein expression of M1-type macrophage factors iNOS and TNF-α （P<0.01）， decreased protein expression of M2-type macrophage factors Arg-1 and IL-10 （P<0.05，P<0.01）， and upregulated expression levels of TLR4， myeloid differentiation factor 88 （MyD88）， phosphorylated inhibitor of NF-κB （p-IκB）/NF-κB inhibitor （IκB）， phosphorylated NF-κB （p-NF-κB） p65/NF-κB p65， NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， and pro-Caspase-1 （P<0.05， P<0.01）. Compared with the model group， all Buyang Huanwutang-treated groups and inhibitor groups significantly reduced levels of IL-1β， IL-18， and TNF-α （P<0.01）， suppressed the expression of inflammatory factors in RAW264.7 macrophages， decreased cellular ROS expression levels （P<0.01）， downregulated M1-type macrophages iNOS and TNF-α protein expression （P<0.01）， upregulated M2-type macrophages Arg-1 and IL-10 protein expression （P<0.01）， and lowered protein expression levels of TLR4， MyD88， p-IκB/IκB， p-NF-κB p65/NF-κB p65， NLRP3， ASC， and pro-Caspase-1 （P<0.05， P<0.01）. ConclusionBuyang Huanwutang can improve macrophage inflammation， potentially by reducing macrophage ROS levels， inhibiting RAW264.7 macrophage polarization， and downregulating the protein expression levels of the TLR4/NF-κB/NLRP3 pathway.

Nutritional status and its influencing factors in children with newly diagnosed inflammatory bowel disease / 中国当代儿科杂志

OBJECTIVES@#To investigate the nutritional status and its influencing factors in children with newly diagnosed inflammatory bowel disease (IBD).@*METHODS@#A retrospective analysis was conducted on the clinical data of children who were diagnosed with IBD for the first time in Hunan Children's Hospital from January 2015 to December 2021. Diagnostic delay was defined as the time from the symptom onset to IBD diagnosis being in the upper quartile (P76-P100) of all IBD children in the study. Multivariate logistic regression analysis was used to explore the risk factors for emaciation and growth retardation.@*RESULTS@#A total of 125 children with newly diagnosed IBD were included, with Crohn's disease being the main type (91.2%). The rates of emaciation and growth retardation were 42.4% (53 cases) and 7.2% (9 cases), respectively, and the rate of anemia was 77.6% (97 cases). Diagnostic delay was noted in 31 children (24.8%), with the time from the symptom onset to IBD diagnosis of 366 to 7 211 days. Multivariate logistic regression analysis showed that diagnostic delay was a risk factor for emaciation and growth retardation (OR=2.73 and OR=4.42, respectively; P<0.05) and that age was positively associated with emaciation (OR=1.30, P<0.05).@*CONCLUSIONS@#Children with newly diagnosed IBD have poor nutritional status, and the rates of anemia, emaciation, and growth retardation are high. Diagnostic delay is associated with malnutrition in children with IBD.

Intestinal and pharyngeal microbiota in early neonates: an analysis based on high-throughput sequencing / 中国当代儿科杂志

OBJECTIVES@#To investigate the distribution characteristics and correlation of intestinal and pharyngeal microbiota in early neonates.@*METHODS@#Full-term healthy neonates who were born in Shanghai Pudong New Area Maternal and Child Health Hospital from September 2021 to January 2022 and were given mixed feeding were enrolled. The 16S rRNA sequencing technique was used to analyze the stool and pharyngeal swab samples collected on the day of birth and days 5-7 after birth, and the composition and function of intestinal and pharyngeal microbiota were analyzed and compared.@*RESULTS@#The diversity analysis showed that the diversity of pharyngeal microbiota was higher than that of intestinal microbiota in early neonates, but the difference was not statistically significant (P>0.05). On the day of birth, the relative abundance of Proteobacteria in the intestine was significantly higher than that in the pharynx (P<0.05). On days 5-7 after birth, the relative abundance of Actinobacteria and Proteobacteria in the intestine was significantly higher than that in the pharynx (P<0.05), and the relative abundance of Firmicutes in the intestine was significantly lower than that in the pharynx (P<0.05). At the genus level, there was no significant difference in the composition of dominant bacteria between the intestine and the pharynx on the day of birth (P>0.05), while on days 5-7 after birth, there were significant differences in the symbiotic bacteria of Streptococcus, Staphylococcus, Rothia, Bifidobacterium, and Escherichia-Shigella between the intestine and the pharynx (P<0.05). The analysis based on the database of Clusters of Orthologous Groups of proteins showed that pharyngeal microbiota was more concentrated on chromatin structure and dynamics and cytoskeleton, while intestinal microbiota was more abundant in RNA processing and modification, energy production and conversion, amino acid transport and metabolism, carbohydrate transport and metabolism, coenzyme transport and metabolism, and others (P<0.05). The Kyoto Encyclopedia of Genes and Genomes analysis showed that compared with pharyngeal microbiota, intestinal microbiota was more predictive of cell motility, cellular processes and signal transduction, endocrine system, excretory system, immune system, metabolic diseases, nervous system, and transcription parameters (P<0.05).@*CONCLUSIONS@#The composition and diversity of intestinal and pharyngeal microbiota of neonates are not significantly different at birth. The microbiota of these two ecological niches begin to differentiate and gradually exhibit distinct functions over time.

Analysis of refractive status of preschool children in Suzhou High tech Zone / 中国学校卫生

Objective@#To observe the refractive status of preschool children, and to explore the prevalence and development trend of ametropia in preschool children, so as to provide support for controlling adolescent ametropia and improving children s health care service.@*Methods@#This cross sectional survey was conducted among 27 561 preschool children (55 122 eyes) aged 3-6 years old in the High tech Zone of Suzhou from September to December 2020. The refractive status was screened by the US Weilun binocular optometry.@*Results@#The total detection rate of ametropia was 9.5%, including 8.6% of astigmatism, 1.3% of hyperopia and 0.5% of myopia. The detection rates of myopia (1.1%) and hyperopia (2.2%) were the highest in the 6-year-old group, and the older the age was, the lower the spherical diopter was, and the higher the cylindrical diopter was. The abnormal rate of colposcopy in girls ( 1.3% ) was higher than that in boys (0.9%), and the abnormal rate of total anisometropia in women (2.3%) was also higher than that in men (1.9%). The main astigmatism was mixed astigmatism (49.1%) and compound hyperopia astigmatism (39.2%); The older the age, the lower the detection rate of compound hyperopia astigmatism. And it is dominated by regular astigmatism( 97.5% ); The higher the age, the higher the detection rate of astigmatism with the rule, while the lower the detection rates of astigmatism against the rule and oblique axis astigmatism.@*Conclusion@#The detection rate of myopia and hyperopia increased significantly at the age of 6, and anisometropia and axial astigmatism also reached the highest at the age of 6. Local health care departments should pay attention to children s astigmatism, especially astigmatism with the rule.

Clinical features of intestinal polyps and risk factors for secondary intussusception in children: an analysis of 2 669 cases / 中国当代儿科杂志

OBJECTIVES@#To study the clinical features of intestinal polyps and the risk factors for secondary intussusception in children.@*METHODS@#A retrospective analysis was performed for the medical data of 2 669 children with intestinal polyps. According to the presence or absence of secondary intussusception, they were divided into two groups: intussusception (n=346) and non-intussusception (n=2 323). Related medical data were compared between the two groups. The multivariate logistic regression analysis was used to identify the risk factors for secondary intussusception.@*RESULTS@#Among the children with intestinal polyps, 62.42% were preschool children, and the male/female ratio was 2.08∶1; 92.66% had hematochezia as disease onset, and 94.34% had left colonic polyps and rectal polyps. There were 346 cases of secondary intussusception, with an incidence rate of 12.96% (346/2 669). Large polyps (OR=1.644, P<0.001), multiple polyps (≥2) (OR=6.034, P<0.001), and lobulated polyps (OR=93.801, P<0.001) were the risk factors for secondary intussusception.@*CONCLUSIONS@#Intestinal polyps in children often occur in preschool age, mostly in boys, and most of the children have hematochezia as disease onset, with the predilection sites of the left colon and the rectum. Larger polyps, multiple polyps, and lobulated polyps may increase the risk of secondary intussusception, and endoscopic intervention is needed as early as possible to improve prognosis.

IRE1α deficiency impairs autophagy in chondrocytes by upregulating calcium homeostasis endoplasmic reticulum protein / 南方医科大学学报

OBJECTIVE@#To explore the mechanism by which inositol-requiring enzyme-1α (IRE1α) regulates autophagy function of chondrocytes through calcium homeostasis endoplasmic reticulum protein (CHERP).@*METHODS@#Cultured human chondrocytes (C28/I2 cells) were treated with tunicamycin, 4μ8c, rapamycin, or both 4μ8c and rapamycin, and the expressions of endoplasmic reticulum (ER) stress- and autophagy-related proteins were detected with Western blotting. Primary chondrocytes from ERN1 knockout (ERN1 CKO) mice and wild-type mice were examined for ATG5 and ATG7 mRNA expressions, IRE1α and p-IRE1α protein expressions, and intracellular calcium ion content using qPCR, Western blotting and flow cytometry. The effect of bafilomycin A1 treatment on LC3 Ⅱ/LC3 Ⅰ ratio in the isolated chondrocytes was assessed with Western blotting. Changes in autophagic flux of the chondrocytes in response to rapamycin treatment were detected using autophagy dual fluorescent virus. The changes in autophagy level in C28/I2 cells overexpressing CHERP and IRE1α were detected using immunofluorescence assay.@*RESULTS@#Tunicamycin treatment significantly up-regulated ER stress-related proteins and LC3 Ⅱ/LC3 Ⅰ ratio and down-regulated the expression of p62 in C28/I2 cells (P < 0.05). Rapamycin obviously up-regulated LC3 Ⅱ/LC3 Ⅰ ratio (P < 0.001) in C28/I2 cells, but this effect was significantly attenuated by co-treatment with 4μ8c (P < 0.05). Compared with the cells from the wild-type mice, the primary chondrocytes from ERN1 knockout mice showed significantly down-regulated mRNA levels of ERN1 (P < 0.01), ATG5 (P < 0.001) and ATG7 (P < 0.001), lowered or even lost expressions of IRE1α and p-IRE1α proteins (PP < 0.01), and increased expression of CHERP (P < 0.05) and intracellular calcium ion content (P < 0.001). Bafilomycin A1 treatment obviously increased LC3 Ⅱ/ LC3 Ⅰ ratio in the chondrocytes from both wild-type and ERN1 knockout mice (P < 0.01 or 0.05), but the increment was more obvious in the wild-type chondrocytes (P < 0.05). Treatment with autophagy dual-fluorescence virus resulted in a significantly greater fluorescence intensity of LC3-GFP in rapamycin-treated ERN1 CKO chondrocytes than in wild-type chondrocytes (P < 0.05). In C28/I2 cells, overexpression of CHERP obviously decreased the fluorescence intensity of LC3, and overexpression of IRE1α enhanced the fluorescence intensity and partially rescued the fluorescence reduction of LC3 caused by CHERP.@*CONCLUSION@#IRE1α deficiency impairs autophagy in chondrocytes by upregulating CHERP and increasing intracellular calcium ion content.

Reflections on integrating humanistic quality education into surgery teaching / 中华医学教育探索杂志

In view of the problems existing in the educational philosophy of medical colleges in China, such as the separation of humanistic knowledge and medical knowledge, the small amount of humanities courses in curriculum arrangement, and the monotonous and boring teaching method of outdated teaching content, this study attempts to optimize the curriculum and increase the proportion of humanities courses; update teaching methods to allow new technologies and new ideas to activate the classroom; strengthen the application of humanistic knowledge in practice according to the characteristics of surgery teaching, so that students can experience humanistic quality education in a real environment; create a good atmosphere for the cultivation of humanistic quality of surgeons, so that students can always feel the edification of humanistic spirit.

Protective effect of rutin on myelin sheath injury in the sciatic nerve of rats induced by acrylamide / 解剖学报

[Abstract] Objective To investigate the effect of rutin (Rut) on sciatic nerve myelin injury induced by acrylamide(ACR), and to observe the changes of myelin structure, myelin basic protein (MBP) and myelin associated glycoprotein (MAG) in rats exposed to ACR. Methods Thirty-six adult male SD rats were randomly divided into 4 groups: control group (ddH

Research on the Development Trend of Pharmaceutical Intermediates Application Technology in China from the Perspective of Patent / 中国药房

OBJECTIVE：To assist enterprises to understand the technical points a nd development trend of pharmaceutical intermediates industry ，and to provide new development ideas and technical support for pharmaceutical intermediates application technology. METHODS ：Taking the core technology involved in pharmaceutical intermediates as the research object and divided into four levels ，the patent data of pharmaceutical intermediates published （announced）before Feb. 2nd，2020 in State Intellectual Property Office （SIPO），INCOPAT and PatSnap patent databases were searched. Patent analysis method was used to analyze the patent application trend and patent applicants of pharmaceutical intermediates ，and the secondary and tertiary technologies of pharmaceutical intermediates were analyzed in detail. Finally ，SPSS 24.0 software was used for curve fitting prediction analysis of the number of patent applications for pharmaceutical intermediates in China. RESULTS ：A total of 12 103 related patents were retrieved. Under the secondary technology classification ，there were 331 patents for antibiotic drug intermediates ，474 patents for antipyretic and analgesic drug intermediates ，1 227 patents for cardiovascular drug intermediates ，and 1 550 patents for anticancer drug intermediates. The numbers of patent applications for pharmaceutical intermediates in the world and China were increasing year by year ，and Chinese mainland had the largest number of patent applications. The number of patent applications in Jiangsu ， Shanghai and Shandong ranked the top 3. Among the top 10 patent，8 were domestic enterprises or universities ，and they also ranked the top 2 in the composition of pharmaceutical intermediates applicants in China. Among the secondary technologies of pharmaceutical intermediates ，cardiovascular pharmaceutical intermediates accounted for the largest proportion of patent applications （32％），followed by anticancer pharmaceutical intermediates （30％）. Among the tertiary technologies ，there were patent applications for the products （35％），methods（35％）and uses （28％）of pharmaceutical intermediates. The curve fitting of the number of patent applications for pharmaceutical intermediates in China showed that the current trend of patent applications was the best fitted with the cubic curve model ，and it was predicted that the number of patent applications in 2020 and 2021 will be 688 and 781 respectively. CONCLUSIONS ：The technology of pharmaceutical intermediates has developed rapidly in China ，and universities and enterprises have considerable R&D strength. In addition to expanding in technological innovation ，enterprises can also consider the development idea of school-enterprise cooperation and patent cross licensing among enterprises to help the better deve lopment of China ’s pharmaceutical intermediates industry.

Drainage procedure for pancreatolithiasis: re-examination of the pancreatic duct diameter standard

Purpose@#Pancreatic duct decompression relieves pancreatic duct stone (PDS)-associated abdominal pain, though a consensus indication for the drainage procedure of the main pancreatic duct (MPD) is lacking. Moreover, major prognostic factors for postsurgical long-term pain relief and recurrence are largely unknown. @*Methods@#The clinical outcomes of 65 consecutive PDS patients undergoing surgery from 2008–2012 with 3+ years of follow-up were assessed. @*Results@#At postsurgical follow-up (median, 4.5 years; range, 3–7 years; procedure: Partington, n = 32; Frey, n = 27; pancreatoduodenectomy, n = 3; distal pancreatectomy, n = 3), the early complication and complete stone clearance rates were 29.2% and 97%, respectively. Long-term, complete and partial pain relief were 93.9%, 83.1%, and 10.8%, respectively. The risk of pancreatic fistula was higher in the 8 mm group (P < 0.05), and 80% of the pancreatic fistula cases occurred in the <8 mm group. A shorter pain duration (P = 0.007), smaller MPD diameter (P = 0.04), and lower Izbicki pain score (P < 0.001) predicted long-term pain relief. Pain recurrence after initial remission occurred in 5 patients and was only related to pain duration (P = 0.02). Stone recurrence and pancreatic exocrine functional and endocrine functional deterioration occurred in 2, 5, and 11 patients, respectively. @*Conclusion@#Surgery provides excellent stone clearance, long-term pain relief, and acceptable postoperative morbidity. Using 8 mm as the criterion for drainage surgery can minimize the postoperative pancreatic fistula risk. Individualized and timely surgical treatment may improve the effect of surgery.

Effect of Puerariae Lobatae Radix on Expression of Endoplasmic Reticulum Stress Related Proteins GRP78 and ATF6 in T2DM Rats / 中国实验方剂学杂志

Objective:To observe the effect of Puerariae Lobatae Radix on type 2 diabetes mellitus (T2DM) rats and related mechanism. Method:Ninety SD rats were divided into normal group, model group, metformin group, Puerariae Lobatae Radix high dose group, Puerariae Lobatae Radix medium dose group and Puerariae Lobatae Radix low dose group, 15 rats in each group. The rats in abnormal group were fed with high-fat and high sugar diet for 4 weeks, and then T2DM model was established by intraperitoneal injection of 40 mg·kg-1 streptozotocin (STZ). Puerariae Lobatae Radix high-dose group was intragastrically administered with 2.1 g·kg-1 of Puerariae Lobatae Radix extract powder, Puerariae Lobatae Radix medium-dose group was intragastrically administered with 1.4 g·kg-1 of Puerariae Lobatae Radix extract powder, Puerariae Lobatae Radix low-dose group was intragastrically administered with 0.7 g·kg-1 of Puerariae Lobatae Radix extract powder, 0.2 g·kg-1 of metformin hydrochloride in metformin group, distilled water once a day in normal group and model group. After 8 weeks, fasting blood glucose (FBG), glycosylated serum protein (GSP), insulin (FINS), triglyceride (TG), cholesterol (TC) were measured, and insulin resistance index （HOMA-IR） was calculated. The expression of tumor necrosis factor -α(TNF-α) was observed by immunohistochemistry. Western blot was used to detect the protein expression of glucose regulatory protein 78 （GRP78）, activated transcription factor 6 （ATF6） in pancreatic tissue. Result:Compared with normal group, the contents of FBG, GSP, TG, TC, FINS, TNF-α in model group were significantly increased (P<0.05), the HOMA-IR was significantly increased (P<0.05), the protein expressions of GRP78 and ATF6 in pancreatic tissue were significantly increased (P<0.05). Compared with model group, the contents of FBG, GSP, TG, TC, FINS and TNF-α in the metformin group and Puerariae Lobatae Radix high, medium and low dose groups were significantly decreased (P<0.05), HOMA-IR decreased significantly (P<0.05), and the expression of GRP78 and ATF6 protein in pancreatic tissue decreased significantly (P<0.05). Conclusion:Puerariae Lobatae Radix can significantly improve insulin resistance in T2DM rats, inhibit the expression of inflammatory factor TNF-α in pancreatic tissue, reduce the protein expression of GRP78 and ATF6 in pancreatic tissue.

Effects of hypoxic exosomes on the proliferation, migration and invasion of hepatocellular carcinoma Huh7 cells / 中华肝脏病杂志

Objective@#To explore the effects of hypoxic exosomes secreted from hepatocellular carcinoma Huh7 cells on the proliferation, migration and invasion under co-cultured normoxic condition.@*Methods@#Hypoxic exosomes secreted from Huh7 cells under hypoxic conditions were extracted by differential ultracentrifugation. Transmission electron microscopy, nanoparticles tracking analysis and western blot were used for the identification of hypoxic exosomes. Hypoxic exosomes were co-cultured with Huh7 cells under normoxic conditions. CCK8, cell scratch and transwell assay were used to detect the changes of cell proliferation, migration and invasion. Statistical analysis was performed by one-way ANOVA and t-test.@*Results@#Hypoxic exosomes secreted from Huh7 cells ranged in size from 30 to 150 nm in diameter, and expressed exosome surface markers CD9, CD63 and TSG101. Hypoxic exosomes significantly enhanced the proliferation of normoxic Huh7 cells (A value of hypoxic exosomes and control group at 48 and 72 h were 2.131 ± 0.092 and 1.760 ± 0.104,t= 3.740,P<0.01, 3.121 ± 0.157 and 2.298 ± 0.085,t= 8.289,P< 0.01). The migration distance between hypoxic exosome and control group at 48 and 72 h were (0.37 ± 0.06 cm)and(0.19 ± 0.05 cm),t= 4.813,P< 0.05, (1.15 ± 0.07 cm) and(0.62 ± 0.08 cm),t= 8.874,P< 0.05, and invasion ability [hypoxic exosomes and control group were (123 ± 18), (44 ± 12),t= 6.203,P< 0.01].@*Conclusion@#Hypoxic exosomes secreted from hepatocellular carcinoma Huh7 cells can promote cell proliferation, migration and invasion in hypoxic environment, suggesting that intercellular information transmission mediated by hypoxic exosomes may be one of the key mechanisms for the amplification of malignancy of hepatocellular carcinoma cells in hypoxic microenvironment.

Effect of Shenling Baizhu San on Murine Model of IBD Induced by DSS in Mice by Regulating Autophagy / 中国实验方剂学杂志

Objective: To investigate the mechanism of Shenling Baizhu San (SLBZS) in treating dextra sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) mice and its relationship with autophagy. Method: SPF BALB/c mice were randomly divided into control group,model group,mesalazine group,low,medium and high-dose SLBZS groups,and autophagy inducer rapamycin group.The IBD mice were fed with 5% DSS in their drinking water for 7 days,and the control mice received only water.SLBZS groups were given SLBZS at doses of 3,6,12 g·kg-1·d-1, positive group was given mesalazine sustained release granules at the dose of 2 g·kg-1·d-1, rapamycin group was given rapamycin at the dose of 4 mg·kg-1·d-1, and control mice was given the same volume of normal saline by gavage.The mice weight,stool occult blood in stool,score of disease activity (DAI),pathological examination of intestinal mucosal lesions integral were observed after 7 days. interleukin(IL)-8 and IL-10 in serum were detected by enzyme\|linked immuno sorbent assay(ELISA), vascular tissue samples were prepared for the detection of tumor neorosis factor-(TNF-α) and IL-1β, and transmission electron microscope and Western blot were used to detect the formation of autophagosomes and the level of autophagy. Result: The body mass decrease, the colon length, disease activity scoring, and histological scoring of SLBZS group were better than those of DSS group. Compared with control group, the level of IL-10 decreased, while the level of IL-8 increased obviously (Pα expressions significantly up-regulated(PPPα expressions(PConclusion: Shenling Baizhu San can significantly inhibit the IBD by regulating autophagy and suppressing inflammation.

Effect of Shenling Baizhu San on Murine Model of IBD Induced by DSS in Mice Through Regulating of Autophagy / 中国实验方剂学杂志

Objective:To investigate the effect of Shenling Baizhu San on inflammatory bowel disease (IBD) induced by 5% dextran sodium sulfate (DSS) in mice by regulating autophagy of intestinal epithelial cells. Method:The 84 BALB/c mice were randomly divided into 7 groups with 12 mice in each group. In addition to the normal group, 5% DSS was freely drunk for 7 days to induce acute inflammatory bowel disease. In the treatment group, high, medium and low doses of shenlingbaishu powder (12,6,3 g·kg-1·d-1), mesalazine (2 g·kg-1·d-1) and autophagy inducer rapamycin (4 mg·kg-1·d-1) were given by gavage. Hematoxylin-eosin (HE) staining was used to observe the morphological changes of colon tissues in mice. The autophagosome formation of intestinal epithelial cells was detected by transmission electron microscopy. Western blot was used to detect the ratio of autophagy related protein LC3-Ⅱ/LC3-Ⅰ, phosphatidylinositol-3kinase (PI3K), mammalian target of rapamycin (mTOR), ubiquitin-binding protein-1 (p62), Beclin1 phosphorylation, ULK1, 4EBP protein expression. Result:Compared with normal group, model group mice colonic mucosa epithelial cells are widely missed, most incomplete glands, change in colitis, LC3-Ⅱ content decreased significantly (PPPPPConclusion:The effect of Shenling Baizhu San on DSS-induced IBD is related to the regulation of the phosphorylation of PI3K, mTOR and p62 proteins in the autophagy pathway of intestinal epithelial cells.

Effect of Buyang Huanwu Tang on LPS-induced Macrophage Activation and Autophagy / 中国实验方剂学杂志

Objective:To investigate the effect of Buyang Huanwu Tang in resisting lipopolysaccharide (LPS)-induced macrophage activation and autophagy through phosphatidylinositol 3-kinase/protein kinase B/mammalian rapamycin target protein (PI3K/Akt/mTOR) signaling pathway. Method:The cell counting kit-8 (CCK-8) method was used to screen out the optimal LPS concentration for inducing the activity of RAW264.7 macrophages. RAW264.7 macrophages were treated separately with PI3K blocker 3-methyladenine(3-MA) (5 mmol·L-1), Akt blocker MK2206 (5 μmol·L-1), mTOR blocker Rapamycin (10 μmol·L-1), Beclin1 blocker Spautin-1 (5 μmol·L-1), different doses of Buyang Huanwu Tang serum (5%, 10%, 20%) and the optimum concentration of LPS for 24 h. The concentrations of inflammatory factors interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in RAW264.7 macrophages were detected by enzyme-lined immunosorbent assay(ELISA). Western blot was used to detect the expression levels of phosphorylated PI3K, phosphorylated Akt, phosphorylated mTOR protein, microtubule light chain protein 3 (LC3), ubiquitin-binding protein 1 (p62) and Beclin-1. The autophagy flow of RAW264.7 cells was detected by transfection with autophagy double-labeled adenovirus. Result:Results of CCK-8 showed the highest cell viability when 10 mg·L-1 LPS was applied. The concentrations of IL-1β, IL-6 and TNF-α in the model group were significantly higher than those in the blank group (PPβ, IL-6 and TNF-α (PPPPPPPPConclusion:Buyang Huanwu Tang can resist LPS-induced macrophages activation and autophagy, inhibit macrophage inflammatory response, regulate PI3K/Akt/mTOR signaling pathway, and inhibit the excessive occurrence of autophagy.

Study on the Optimization of the Extraction Technology of Shengmaiyin Polysaccharide and Its Regulation Effects on Intest- inal Function of Spleen Deficiency Model Rats / 中国药房

OBJECTIVE： To optimize the extraction technology of Shengmaiyin polysaccharide， and to investigate the regulation effects of Shengmaiyin and its polysaccharide on intestinal function of spleen deficiency model rats. METHODS： The contents of polysaccharide were determined by phenol-sulfuric acid method， and the extraction rate of polysaccharide was calculated. Using extraction rate of Shengmaiyin polysaccharide as investigation index， singel factor and orthogonal tests were used to optimize material-liquid ratio， extraction time， extraction temperature and extraction times of Shengmaiyin polysaccharide. Validation test was also conducted. Totally 80 male SD rats were randomly divided into blank group， model group， Shengmaiyin low-dose， medium-dose and high-dose groups （350， 700， 1 400 g/L， by crude drug）， Shengmaiyin polysaccharide low-dose， medium-dose and high-dose groups （24.5， 49， 98 g/L， by crude drug）， with 10 rats in each group. Except for blank group， other groups were given Rheum palmatum water decoction 10 mL/kg to induce spleen deficiency model， once a day， for consecutive 15 d. Since the 16th day， blank group and model group were given isovolumic water intragastrically， while other groups were given corresponding drugs， once a day， for consecutive 10 d. The general status of rats and body weights were recorded in each group. The serum contents of D-xylose， gastrin （GAS） and vasoactive intestinal peptide （VIP） were detected by phloroglucinol method or ELISA. RESULTS： The optimal extraction technology of Shengmaiyin polysaccharide was material-liquid ratio 1 ∶ 10（g/mL）， extraction time 45 min， extraction temperature 80 ℃， extracting for 1 time. Results of validation test showed that extraction rates of the polysaccharide in 3 times were 7.43％， 7.64％， 7.80％ （RSD＝1.01％， n＝3）. After modeling， except for blank group， other groups suffered from loose stools， thin body and reduced food intake， and the body weight and serum level of D-xylose were decreased significantly compared with blank group （P＜0.01）. After last medication， above symptoms of administration groups were improved to different extents. Except for model group， body weight and serum contents of D-xylose in other groups were increased significantly than those before modeling or before medication （P＜0.05 or P＜0.01）. Compared with blank group， body weight and serum content of GAS were decreased significantly in model group， while serum content of VIP was increased significantly （P＜0.01）. Compared with model group， body weight of Shengmaiyin medium-dose group and Shengmaiyin polysaccharide low-dose and high-dose groups， serum contents of D-xylose and GAS in Shengmaiyin medium-dose and high-dose groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were increased significantly， while serum contents of VIP in Shengmaiyin groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： The optimized extraction technology of Shengmaiyin polysaccharide is stable and feasible. Shengmaiyin and its polysaccharide contribute to the recovery of intestinal function of spleen deficiency model rat， the effects of which may be associated with the secretion regulation of GAS and VIP.

In vitro evaluation of aconitine-induced arrhythmia based on RTCA Cardio technology / 中国药理学通报

Aim To establish an in vitro arrhythmia detection technique based on real-time cell analysis (RTCA) Cardio sys-tem with aconitine as a tool drug, and to provide a reliable method for the development of antiarrhythmic drugs. Methods The effects of aconitine on rat cardiac rhythm were detected by eight-channel physiological recorder at the level of whole animal. In vitro cultured cardiac myocytes, the inoculation density of cardiac myocytes was investigated by HTCA method, the effect of aconitine on cardiac beating was monitored by RTCA Cardio system, and the CI value, beating rate, amplitude and irregular rhythm of cardiac myocytes were analyzed. Results Eight-channel physiological recorder was used to detect the effects of aconitine on whole animals. The results showed that aconitine(50 mg • g"1) could induce arrhythmias such as ventricular tachycardia, ventricular fibrillation, shortened RR interval and increased heart rate. RTCA Cardio system showed that aconitine (2-8 p,M) could induce arrhythmias such as increased cardiac cell beating frequency, decreased beating amplitude and abnormal beating state in a dose-dependent manner. Conclusions RTCA Cardio system can rapidly, sensitively and accurately detect the arrhythmia induced by aconitine in cardiac myocytes, which provides methodological reference for the development of antiarrhythmic drugs.