Effective substances and mechanism of Yishen Guluo Mixture in treatment of chronic glomerulonephritis based on metabolomics and serum pharmacochemistry / 中国中药杂志

This study aimed to investigate the effective substances and mechanism of Yishen Guluo Mixture in the treatment of chronic glomerulonephritis(CGN) based on metabolomics and serum pharmacochemistry. The rat model of CGN was induced by cationic bovine serum albumin(C-BSA). After intragastric administration of Yishen Guluo Mixture, the biochemical indexes related to renal function(24-hour urinary protein, serum urea nitrogen, and creatinine) were determined, and the efficacy evaluations such as histopathological observation were carried out. The serum biomarkers of Yishen Guluo Mixture in the treatment of CGN were screened out by ultra-performance liquid chromatography-quadrupole time-of-flight/mass spectrometry(UPLC-Q-TOF-MS) combined with multivariate statistical analysis, and the metabolic pathways were analyzed. According to the mass spectrum ion fragment information and metabolic pathway, the components absorbed into the blood(prototypes and metabolites) from Yishen Guluo Mixture were identified and analyzed by using PeakView 1.2 and MetabolitePilot 2.0.4. By integrating metabolomics and serum pharmacochemistry data, a mathematical model of correlation analysis between serum biomarkers and components absorbed into blood was constructed to screen out the potential effective substances of Yishen Guluo Mixture in the treatment of CGN. Yishen Guluo mixture significantly decreased the levels of 24-hour urinary protein, serum urea nitrogen, and creatinine in rats with CGN, and improved the pathological damage of the kidney tissue. Twenty serum biomarkers of Yishen Guluo Mixture in the treatment of CGN, such as arachidonic acid and lysophosphatidylcholine, were screened out, involving arachidonic acid metabolism, glycerol phosphatide metabolism, and other pathways. Based on the serum pharmacochemistry, 8 prototype components and 20 metabolites in the serum-containing Yishen Guluo Mixture were identified. According to the metabolomics and correlation analysis of serum pharmacochemistry, 12 compounds such as genistein absorbed into the blood from Yishen Guluo Mixture were selected as the potential effective substances for the treatment of CGN. Based on metabolomics and serum pharmacochemistry, the effective substances and mechanism of Yishen Guluo Mixture in the treatment of CGN are analyzed and explained in this study, which provides a new idea for the development of innovative traditional Chinese medicine for the treatment of CGN.

Serum osteocalcin and diabetic peripheral neuropathy in male patients with diabetes / 中华内分泌代谢杂志

Objective:To investigate the association between serum bone turnover marker osteocalcin and the distal symmetric poly neuropathy(DSPN) in male diabetic patients.Methods:Clinical data from 370 male diabetic patients who admitted to Zhongshan Hospital, Fudan University from January 2020 to November 2021 were collected. These patients were grouped into tertiles by serum osteocalcin level: T1 group(osteocalcin<9.2 ng/mL, n=123), T2 group(osteocalcin 9.2-13 ng/mL, n=122), and T3 group(osteocalcin≥13 ng/mL, n=125). The percentage ratios of DSPN were compared among these groups. Using logistic regression model, the adjusted odds ratio ( OR) for DSPN was calculated. Results:There were 50(40.7%), 29(23.8%), 49(39.2%) patients with DSPN in T1, T2, and T3 group respectively. The ratio of patients with DSPN in osteocalcin T2 group were lower than that in the T1 and T3 groups. Further logistic regression showed a 133.9%( OR=2.339, 95% CI 1.097-4.988, P=0.028) and a 134.2%( OR= 2.342, 95% CI 1.040-5.275, P=0.039) increased risk for DSPN in the T1 and T3 group respectively compared with the T2 group, even after adjusted for age, diabetic duration, HbA 1C, diabetic complications, β cross-linked C-telopeptide of type Ⅰ collagen(CTXβ), 25-hydoxy vitamin D(25-OHD), bone mineral density, and treatment. Conclusions:The serum levels of bone turnover marker osteocalcin were associated with the occurrence of DSPN in male diabetic patients, a moderate level of bone turnover(the serum osteocalcin level of between 9.2 and 13 ng/mL for instance) might be protective for male diabetic patients from DSPN.

Study on the mechanism of Wenyang huazhuo tongluo formula contained serum inhibiting the proliferation of Th17 cells in scleroderma patients / 中国药房

OBJECTIVE To study the effects of Wenyang huazhuo tongluo formula conta ined serum on the expression and methylation of retinoic acid related nuclear orphan receptor （RORγt），and to explore the mechanism of its regulation of Th 17 cell proliferation in the treatment of scleroderma. METHODS Wistar rats were given 15，30，60 g/（kg·d）Wenyang huazhuo tongluo formula intragastrically to prepare different doses of drug-contained serum ，and peripheral blood of scleroderma patients were collected to sort Th 17 cells. Using blank serum as blank control ，methyltransferase inhibitor decitabine （DCA）as positive control ， Th17 cells were treated with different concentrations of drug-contained serum ，and then the proliferation of Th 17 cells was detected by CCK- 8；mRNA expressions of RORγt and IL-17 were detected by qRT-PCR ，and the protein expression of RORγt was detected by Western blot assay ；the protein expression of IL- 17 in the cell supernatant was detected by ELISA ，and the methylation level of RORγt gene promoter was detected by methylation-specific PCR. The transcriptional activity of RORγt gene promoter was detected by dual-luciferase assay. RESULTS Compared with blank control ，Wenyang huazhuo tongluo formula contained serum and DCA could inhibit the proliferation of Th 17 cells（P＜0.05），reduced the mRNA and protein expressions of RORγt and IL-17，enhanced the methylation level of RORγt gene promoter and attenuated ， the transcription activity of RORγt gene promoter. Except for mRNA expression of Th 17 and the methylation level of RORγt gene promoter in drug-contained serum low-dose group ，there were statistical significance in above indexes of other groups（P＜0.05）. CONCLUSIONS Wenyang huazhuo tongluo formula can promote the methylation lev el of RORγt gene，and inhibit the expression of RORγt and the secretion of IL-17，so as to inhibit the proliferation of Th 17 cells.

Effective substance and mechanism of Ziziphi Spinosae Semen extract in treatment of insomnia based on serum metabolomics and network pharmacology / 中国中药杂志

This study aims to study the effective substance and mechanism of Ziziphi Spinosae Semen extract in the treatment of insomnia based on serum metabolomics and network pharmacology. The rat insomnia model induced by p-chlorophenylalanine(PCPA) was established. After oral administration of Ziziphi Spinosae Semen extract, the general morphological observation, pentobarbital sodium-induced sleep test, and histopathological evaluation were carried out. The potential biomarkers of the extract in the treatment of insomnia were screened by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS) combined with multivariate analysis, and the related metabolic pathways were further analyzed. The "component-target-pathway" network was constructed by ultra-high performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry(UHPLC-Q-Exactive-MS/MS) combined with network pharmacology to explore the effective substances and mechanism of Ziziphi Spinosae Semen in the treatment of insomnia. The results of pentobarbital sodium-induced sleep test and histopathological evaluation(hematoxylin and eosin staining) showed that Ziziphi Spinosae Semen extract had good theraputic effect on insomnia. A total of 21 endogenous biomarkers of Ziziphi Spinosae Semen extract in the treatment of insomnia were screened out by serum metabolomics, and the metabolic pathways of phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, and nicotinate and nicotinamide metabolism were obtained. A total of 34 chemical constituents were identified by UHPLC-Q-Exactive-MS/MS, including 24 flavonoids, 2 triterpenoid saponins, 4 alkaloids, 2 triterpenoid acids, and 2 fatty acids. The network pharmacological analysis showed that Ziziphi Spinosae Semen mainly acted on target proteins such as dopamine D2 receptor(DRD2), 5-hydroxytryptamine receptor 1 A(HTR1 A), and alpha-2 A adrenergic receptor(ADRA2 A) in the treatment of insomnia. It was closely related to neuroactive ligand-receptor interaction, serotonergic synapse, and calcium signaling pathway. Magnoflorine, N-nornuciferine, caaverine, oleic acid, palmitic acid, coclaurine, betulinic acid, and ceanothic acid in Ziziphi Spinosae Semen may be potential effective compounds in the treatment of insomnia. This study revealed that Ziziphi Spinosae Semen extract treated insomnia through multiple metabolic pathways and the overall correction of metabolic disorder profile in a multi-component, multi-target, and multi-channel manner. Briefly, this study lays a foundation for further research on the mechanism of Ziziphi Spinosae Semen in treating insomnia and provides support for the development of innovative Chinese drugs for the treatment of insomnia.

Syndrome Distribution of 9 323 Cough Cases in Ancient Chinese Medical Books Based on Latent Structure Model and Logistic Regression Analysis / 中国实验方剂学杂志

Objective:To explore the syndrome distribution of cough cases in ancient Chinese medical books. Method:A total of 9 323 cough cases in the database of lung diseases in ancient Chinese medical books were extracted. Lantern 5.0 was used to construct the latent structure model for the 100 manifest variables based on the first 50 symptoms and 50 Chinese herbal medicines， and different latent nodes were interpreted. The syndrome differentiation patterns of syndromes with symptoms and Chinese herbal medicine （formula） were quantitatively revealed by the comprehensive clustering weights of latent structure. The correlation of diseases with syndromes was fitted through the binary Logistic regression analysis. Result:There were 204 syndromes involved in 9 323 cough cases with 18 syndromes showing a frequency higher than 100. As demonstrated by the model established on the first 50 symptoms and 50 Chinese herbal medicines， 35 latent variables， 98 latent classes， and 10 comprehensive clustering models were obtained， where <italic>Z</italic>5<italic> </italic>was the highest in the threshold value （6.7）， <italic>Z</italic>6 in the information coverage of productive cough （52%）， and <italic>Z</italic>7 in the score （19）. The binary Logistic regression model fitted the correlation between different disease types and five syndromes， where the dominance ratio of external wind to the syndrome of wind-heat invading lung reaching up to 88.919， those of syndrome of phlegm-heat accumulating in lung to diseased heat and sputum 51.594 and 15.861， and those of the syndrome of phlegm-dampness obstructing lung to dampness， phlegm， and fluid retention 31.415， 34.370， and 4.936， respectively. Conclusion:The newly discovered cough syndromes included lung heat and yin deficiency， external cold and internal fluid retention， weakness of spleen and stomach， and phlegm and blood stasis in lung. In most cases， multiple syndromes were observed clinically， such as syndrome of deficiency of both Qi and Yin in lung combined with yin deficiency in lung and kidney. Since differentiation of traditional Chinese medicine （TCM） syndrome is subjective， the weight can indicate the difference in the contributions of different symptoms to the syndrome， which is of guiding significance for syndrome inference. The latent structure model combined with Logistic regression analysis can solve the problem of quantification in TCM syndrome differentiation and can be used to explore the syndrome distribution of diseases.

Spectrum-effect relationship of the blood-activating efficacy of Notoginseng Radix et Rhizoma / 中国中药杂志

This study aims to establish the high-performance liquid chromatography(HPLC) fingerprints of different batches of Notoginseng Radix et Rhizoma, determine their pharmacodynamic indexes of promoting blood circulation, and explore the spectrum-effect relationship between the chemical components of Notoginseng Radix et Rhizoma and the efficacy of promoting blood circulation. Firstly, the HPLC fingerprints of different batches of Notoginseng Radix et Rhizoma were established. Then, the pharmacodynamic indexes were determined after the capillary coagulation experiment and the cerebral ischemia-reperfusion in rats, including capillary coagulation time, percentage of cerebral ischemic area, cerebral water loss rate, and brain-body index. Afterward, the partial least-squares method was used to explore the spectrum-effect relationship between the chemical components of Notoginseng Radix et Rhizoma and the pharmacodynamic indexes. The results showed that this study successfully established the HPLC fingerprints of different batches of Notoginseng Radix et Rhizoma, found 23 common peaks, and identified 12 of them, all of which were saponins. The method was proved stable and reliable. Both the capillary coagulation experiment and the middle cerebral artery occlusion(MCAO)-induced cerebral ischemia-reperfusion experiment on rats revealed that there were obvious differences in the pharmacodynamic indexes of different batches of Notoginseng Radix et Rhizoma. The relationships between 23 common components of Notoginseng Radix et Rhizoma in different batches and the pharmacodynamic indexes were discussed by means of spectrum-effect correlation analysis, of which 17 components had positive effects while 6 components had negative effects on the pharmacodynamic indexes. This study provides a certain reference basis for the clinical rational use and quality control of Notoginseng Radix et Rhizoma.

Spectrum-effect relationship of hemostatic effects of Notoginseng Radix et Rhizoma with different commodity specifications / 中国中药杂志

This article aims to establish the fingerprints, determine the hemostatic pharmacodynamic indicators, and explore the spectrum-effect relationship of Notoginseng Radix et Rhizoma in 12 different specifications. Firstly, HPLC and liquid chromatography-mass spectrometry(LC-MS) were employed to establish the fingerprints of Notoginseng Radix et Rhizoma. The rat plasma recalcification experiment and the rat gastric bleeding experiment were conducted to determine the pharmacodynamic indicators, including plasma recalcification time(PRT), thrombin time(TT), prothrombin time(PT), and activated partial thromboplastin time(APTT). Afterwards, the partial least squares method was employed to explore the spectrum-effect relationship of Notoginseng Radix et Rhizoma in different specifications. Twenty-six common peaks were detected in the HPLC fingerprints of different specifications of Notoginseng Radix et Rhizoma, and 11 out of the 26 common peaks represented saponins. The content of dencichine was determined by LC-MS. The rat experiments showed that the pharmacodynamic indicators were significantly different among different specifications of Notoginseng Radix et Rhizoma. The spectrum-effect relationship was explored between 27 common components and pharmacodynamic indicators. Among them, 16 components had positive effects on the pharmacodynamic indicators of Notoginseng Radix et Rhizoma, and 11 exerted negative effects. This study provides a basis for the precision medication and quality control of Notoginseng Radix et Rhizoma.

Association of sex hormone and sex hormone-binding globulin levels in patients with Graves′ disease / 中华内分泌代谢杂志

Objective:To identify the association of sex hormone and sex hormone-binding globulin (SHBG) levels in patients with Graves′ disease.Methods:Between December 2017 and July 2019, 152 patients with Graves′ disease were enrolled in the Department of Endocrinology and Metabolism of Zhongshan Hospital, Fudan University. Parameters such as height, weight, thyroid function, sex hormone, fasting blood glucose, insulin, C-peptide, glycosylated albumin, HbA 1C, and liver function were collected. The associations between SHBG and bioclinical characteristics were analyzed. Results:The glycosylated albumin level was negatively associated with SHBG in all subjects ( β=-0.308, P<0.01), while this association was not significant after the adjustment for thyroid hormones. Male patients had significantly lower SHBG level than female patients ( P<0.01). In male patients, SHBG was associated with luteinizing hormone ( r=0.465, P<0.01), estradiol ( r=0.629, P<0.01), testosterone ( r=0.786, P<0.01). While in female patients, SHBG was also associated with testosterone ( r=0.191, P<0.05). In multivariate linear regression analysis, estradiol was independently associated with SHBG ( β=0.329, P<0.01) in male subjects. Conclusion:The hypogonadism and infertility in Graves′ disease patients could partially be attributed to the association between SHBG and Graves′ disease.

Prescriptions and Syndromes of 9 323 Cough Cases in Ancient Chinese Medical Books Based on Rules of Latent Structure Differentiation / 中国实验方剂学杂志

Objective:To explore cough cases recorded in ancient traditional Chinese medicine (TCM) books based on the Rules of Latent Structure Differentiation. Method:The 9 323 cough cases in the database of ancient TCM books and pulmonary diseases were extracted. At first, Lantern 5.0 software was used to construct the top 120 TCM hidden structure model with the frequency>100. Then, the obtained hidden variables were comprehensively clustered and interpreted. Result:The 46 hidden variables and 114 hidden classes were obtained through the modeling of 120 traditional Chinese medicines. According to the bayesian information measure (BIC) score, the model score was -161 242.92. The model diagram was formed with Y0, Y5, Y7, Y13, Y35 and Y38 as the cores, and Y36, Y38, Y24, Y19 and Y17 involved 4 hidden classes. Ten comprehensive clustering models were summarized according to the hidden structure flow chart, among which the highest score of Z5 external cold and internal decoction syndrome was 8.4, indicating that the result of syndrome differentiation rules of Z5 had a high degree of support, and the highest mutual information degree and information coverage of Pinellia sinensis were 0.31 and 63%. Z2 had a low score for the syndrome of deficiency of lung qi and Yin, which was -2， and Schisandra had the lowest score of -15, indicating that the syndrome differentiation rules of Z2 was less supportive. Forsythia and Notopterygium both had the highest score of Z6 wind-heat attack lung syndrome and Z10 phlegm and blood-stasis pulmonary syndrome, which was 19.1, indicating that these two TCMs had a great contribution to it. According to the test, the common syndromes of cough were cold and dryness attacking the lung, kidney-Yin deficiency, kidney-Yin deficiency of the lung, cold and internal decoction, wind heat invading the lung, liver fire invading the lung, lung heat burning, phlegm-dampness blocking the lung, phlegm-stasis blocking the lung. The newly discovered cough syndromes were external cold and internal rheum syndrome and phlegm-stasis blocking the lung syndrome. Conclusion:Based on the syndrome differentiation rules of hidden structure, the improved classification of cough syndromes provides ideas for the demonstration of tacit knowledge of TCM and methodological reference for the improvement of syndromes of other diseases, and is conducive to the development of valuable new prescriptions.

Is nonalcoholic fatty liver disease a complication of diabetes? / 临床肝胆病杂志

@# Although nonalcoholic fatty liver disease (NAFLD) is a liver disease, it has attracted more and more attention among endocrinologists. There are complex interactions between the metabolic status of diabetes and the pathology of NAFLD, and they promote the development of each other. In the population with type 2 diabetes mellitus (T2DM), the incidence rate of NAFLD is as high as 57%-80%, and NAFLD patients with diabetes are more likely to develop progressive liver diseases such as nonalcoholic steatohepatitis (NASH) and liver fibrosis. NAFLD also leads to the deterioration of glucose and lipid metabolism disorder in diabetes patients, as well as the development and progression of chronic macrovascular and microvascular complications in diabetes. The above evidence supports the view that NASH should be considered a complication of T2DM. Once a definite diagnosis of T2DM is made in clinical practice, the risk of NASH or progressive fibrosis should be evaluated and intervention should be given as early as possible.

Herbal Textual Research on Genuine Medicinal Materials “Deng Ju” and Quality Evaluation of Nan Yang Chrysanthemi Flos / 中国实验方剂学杂志

Objective: To study the origin of Nanyang Chrysanthemi Flos materia medica and its historical origin, in order to evaluate the quality of Nanyang Chrysanthemi Flos by HPLC method, and define the advantages of Nanyang Chrysanthemi Flos with the origin of Nanyang. Method: Records of Nanyang Chrysanthemi Flos in the "Chinese Medical Code" and related ancient documents were studied to explain the origin and application. The genetic relationship between Nanyang Chrysanthemi Flos and other pieces was revealed in ancient literatures. Then 8 chemical constituents in 14 batches of Chrysanthemi Flos were determined by HPLC multi-components quantitation,and the comprehensive weighted score analysis was performed based on the results. The HPLC fingerprints were established,and the similarity analysis and clustering analysis were made to comprehensive evaluation the quality of Nanyang Chrysanthemi Flos and define the genetic relationship between Nanyang and other pieces at the chemical composition level. Result: The results of the herbal textual research show that Nanyang Chrysanthemi Flos spread from place to place since the Han dynasty,and impact many medicinal chrysanthemums of later generations. HPLC fingerprints similarity and cluster analysis also indicated the genetic relationship between Nanyang Chrysanthemi Flos and other species at the chemical level. The comprehensive score analysis results show that Nanyang Chrysanthemi Flos is of good quality and very suitable for medicinal purposes. Conclusion: Nanyang has been a quality production area of Chrysanthemi Flos since ancient times to present. Nanyang Chrysanthemi Flos is very suitable for medicinal purposes and worthy of promotion.

Advances in metabolic engineering of heterotrophic microorganisms for CO₂ fixation: a review / 生物工程学报

Environmental protection and energy supply are our two major concerns. Greenhouse gases released from energy consumption have serious impact on the environment. CO₂ fixation can be used to convert CO₂ into fuels or chemicals. However, natural carbon-fixing organisms usually have some disadvantages such as slow growth and low carbon fixation efficiency. Enhancing or remodeling CO₂ fixation pathways in model microorganisms can realize CO₂ recycling, which can further increase fuel or chemical production and reduce greenhouse gas emission. This review describes in detail metabolic engineering of CO₂ fixation pathways to improve chemical production and sugar synthesis, elaborates the role of relevant metabolic pathways and key enzymes in CO₂ fixation, introduces the application of electro-biochemical synthesis system, shows the great potential of CO₂ fixation, and prospects the future research direction of CO₂ fixation.

The mechanisms of paeonol reversing multidrug resistance in ovarian cancer SKOV3/DDP cells / 药学学报

The purpose of this research is to investigate the effects and mechanisms of paeonol (PL), a phenolic compound found in many traditional Chinese formulations, on reversing drug resistance in the ovarian cancer resistant SKOV3/DDP cells. The results showed that PL had significant drug-resistant reversal effect on SKOV3/DDP cells. Flow cytometry showed that PL could inhibit P-glycoprotein (P-gp) function in a concentration-dependent manner. Fluorescent quantitative PCR and cell immunofluorescence techniques were used to detect mechanisms of action. Results revealed that both the inhibitory effect on MDR1/P-gp and metadherin (MTDH) expression and the induction effect on phosphatase and tensin homolog (PTEN), by 15, 30, and 60 μmol·L-1 PL, were increased with increased concentrations of PL (P P -1 PL treated group; however, the inhibition or induction effect on MTDH or PTEN protein were only comparable to the 15 μmol·L-1 PL treated group. The present study shows that the effect of PL on SKOV3/DDP cells may be related to the inhibition of P-gp function and expression, the inhibition of MDR1, MTDH expression, and the induction of PTEN expression, all which can provide a theoretical foundation for PL as a drug resistance reversal agent on the treatment of ovarian cancer chemotherapy resistance.

Guizhi Fuling Wan reverses drug resistance by regulating PTEN and MTDH in ovarian cancer SKOV3/DDP Cells / 中国药理学与毒理学杂志

OBJECTIVE To detect the reversal effect of Guizhi Fuling Wan on cisplatin-resistant ovarian cancer SKOV3/DDP cells and its relationship with protein expression of phosphatase and tensin homolog (PTEN) and metadherin (MTDH). METHODS Guizhi Fuling Wan (GFW) concentrated solution was prepared according to the Chinese Pharmacopoeia 2015 edition, Wistar rats were given GFW viagavage at 4 g·kg-1·d-1,8 g·kg-1·d-1,16 g·kg-1·d-1,or given saline as blank control for 5 days.Blood samples were taken and the corresponding drug-containing low-dose sera, medium-dose sear, high-dose sera and blank sera were prepared.The XCELLigence RTCA S16 real-time label-free cell analyzer was used to detect the reversal effect by the sera combined with cisplatin or paclitaxel in SKOV3/DDP cells. Annexin V-FITC and PI double-staining were used to detect the apoptosis-inducing effect of the sera in the cells. RT-qPCR and western blot were used to detect the mRNA and protein expression of PTEN and MTDH after the cells treated with the sera. RESULTS The inhibition rate of low-dose sera against SKOV3/DDP cells was less than 5%.After the low-dose sera combined with cisplatin or pacli-taxel, the IC50 of SKOV3/DDP cells against cisplatin and paclitaxel decreased by 3.01 and 1.79-fold, respectively.The total apoptosis rates induced by the low-dose sera,medium-dose sear,high-dose sera and blank sera in SKOV3/DDP cells were 11.08±0.13,19.42±0.30,24.23±0.31,and 3.21±0.24,respec-tively; there was a significant difference between the groups (P<0.01). RT-qPCR results showed that, compared with the blank serum, the sera can up-regulate the expression of PTEN mRNA and down-regulate the expression of MTDH mRNA in a dose-dependent manner. Western blot results showed that the induction effect to PTEN protein and the inhibition effect to MTDH protein by the sera were gradually enhanced with thesera dose increasement. CONCLUSION The resistance reversal effect of Guizhi Fuling Wan on ovarian cancer SKOV3/DDP cells may be related to the inhibition of MTDH, up-regulation of PTEN and induction of apoptosis, providing with an experiment basis for the applica-tion of Guizhi Fuling Wan as a reversal agent for chemotherapy resistance of ovarian cancer.

Effects of RARG on viability and migration ability of gastric cancer cells / 中国病理生理杂志

AIM:To investigate the effect of retinoic acid receptor gamma(RARG)on the viability and migra-tion ability of gastric cancer cells.METHODS:The expression of RARG in gastric cancer and normal gastric tissues and its correlation with the overall survival rate of gastric cancer patients were analyzed by bioinformatics.The expression of RARG was promoted and inhibited by over-expression plasmid transfection and RNA interference technique in gastric can-cer cells in vitro,respectively.MTT and Transwell assays were used to detect the effect of RARG on the viability and mi-gration ability of gastric cancer cells.The effect of RARG on regulating the Wnt/β-catenin signaling pathway was evaluated by Western blot and TOP/FOP dual-luciferase reporter assay.The protein interaction of RARG and β-catenin was deter-mined by co-immunoprecipitation and immunofluorescence co-localization assay.RESULTS:Over-expression of RARG en-hanced the viability and migration ability of gastric cancer SGC 7901 cells(P<0.05).Knockdown of RARG attenuated the viability and migration ability of gastric cancer MGC-803 cells(P<0.05).At the same time, RARG over-expression in-creased the protein expression levels of β-catenin, c-Myc, cyclin D1, Twist and Snail(P<0.05), and the activity of TOP/FOP dual-luciferase reporter gene(P<0.05).In addition, RARG interacted with β-catenin protein in the gastric cancer cells.CONCLUSION:RARG promotes the viability and migration ability of gastric cancer cells via activating the Wnt/β-catenin signaling pathway,thus playing an important role in the development of gastric cancer.

Promoting cell viability and migration of gastric cancer cells by PRRX2 via activation of Wnt/β-catenin signaling pathway / 中国病理生理杂志

AIM:To study the effect of paired-related homeobox 2(PRRX2)gene on the viability and migra-tion ability of gastric cancer cells,and to analyze the underlying mechanism of regulating Wnt /β-catenin signaling pathway. METHODS:The expression of PRRX2 in gastric cancer and normal gastric tissue and the correlation between PRRX 2 ex-pression in gastric cancer tissues with the overall survival rate of gastric cancer patients were analyzed by bioinformatics. The small interfering RNA(siRNA)and over-expressed plasmids of PRRX2 were transfected into gastric cancer cells MGC-803 and SGC-7901,respectively.MTT assay and Transwell assay were used to detect the viability and migration ability of gastric cancer cells.Western blot and TOPflash/FOPflash dual-luciferase reporter gene assay were used to detect the activi-ty of Wnt/β-catenin signaling pathway.Co-immunoprecipitation was used to detected the interaction between PRRX 2 and β-catenin proteins.RESULTS:Knockdown of PRRX2 attenuated the viability and migration ability of gastric cancer cell line MGC-803(P <0.05).Over-expression of PRRX2 enhanced the viability and migration ability of SGC-7901 cells (P<0.05),increased the protein levels of β-catenin, c-Myc and cyclin D1(P<0.05)and the activity of TOPflash/FOPflash dual-luciferase reporter gene(P<0.05).PRRX2 interacted with β-catenin protein in gastric cancer cells.CON-CLUSION:PRRX2 promotes the viability and migration ability of gastric cancer cells, which may be related to Wnt/β-catenin signaling pathway.

The basal rate characteristics of continuous subcutaneous insulin infusion in adult type 1 diabetes with islet failure / 中华内分泌代谢杂志

Objective To investigate the basal rate characteristics of continuous subcutaneous insulin infusion(CSII)in adult type 1 diabetes with islet failure. Methods Adult type 1 diabetic patients with islet failure, treated by CSII for more than 12 months but with poor glycemic control,were optimized for insulin dose through basal rate verification. The differences of blood glucose levels and insulin dosages were compared before and after CSII dose optimization. Results In 16 cases,HbA1Creduced from(7.6 ± 1.1)% by the time of enrollment to(7.0 ± 0.5)% after dose optimization(P<0.05),the minimal blood glucose level rose from(3.1 ± 1.1)mmol/L to(4.5 ± 0.7) mmol/L(P<0.01),and the maximal blood glucose level dropped from(18.6 ± 2.3)mmol/L to(12.0 ± 1.5)mmol/L (P<0.01). The total daily dosage of CSII reduced by about 17% (P<0.01)and the total basal insulin dosage reduced by about 43%(P<0.01)after dose optimization. The percentage of total basal insulin dosage to total daily dosage reduced from(43.6 ± 9.9)% by the time of enrollment to(29.4 ± 3.5)% after dose optimization(P<0.01). During 24 hours of the day,basal rate at 03: 00-08: 00 reached the peak level(1.3 times of the hourly average basal rate level)and basal rate at 20: 00-24: 00 fell to the valley level(60% of the hourly average basal rate level). Conclusions In adult type 1 diabetic patient with islet failure,the total basal insulin requirement of CSII is about 30% of the total daily dosage,with the peak level at 03: 00-08: 00 and the valley level at 20: 00-24: 00.

Effects of lithium chloride on transforming growth factor beta and connective tissue growth factor in cultured human Tenon's capsule fibroblasts / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To research the effects of lithium chloride on transforming growth factor beta (TGF-β) and connective tissue growth factor (CTGF) in cultured human Tenon capsule fibroblasts (HTFs) and explore its mechanism.METHODS:HTFs were cultured and identified by vimentin staining with immunofluorescence and the morphological characteristics.The experimental group was processed 48h with LiCl in concentration of 80mmol/L, the control group without LiCl.The mRNA expression of TGF-β and CTGF in two groups were analyzed with real-time fluorescent quantitative polymerase chain reaction (real time-qPCR) and the protein expression was detected with Western blot.RESULTS:The cultured HTFs expressed TGF-β and CTGF.The mRNA expression of TGF-β and CTGF significantly decreased compared with the control group(t=20.042, 14.995, P<0.05).the protein expression of TGF-β and CTGF also decreased significantly compared with the control group(t=46.058、12.452, P<0.05)CONCLUSION:The cultured HTFs can express TGF-β and CTGF in mRNA and proteins' level.LiCl can reduce the expression of TGF-β and CTGF both in gene and proteins' level.LiCl has the potential to modulate wound healing for glaucoma filtration surgery.

Prevention, diagnosis and treatment of perioperative complications of bariatric and metabolic surgery / 中华胃肠外科杂志

Surgical operation in treating obesity and type 2 diabetes is popularizing rapidly in China. Correct prevention and recognition of perioperation-related operative complications is the premise of ensuring surgical safety. Familiar complications of the operation include deep venous thrombosis, pulmonary artery embolism, anastomotic bleeding, anastomotic fistula and marginal ulcer. The prevention of deep venous thrombosis is better than treatment. The concrete measures contain physical prophylaxis (graduated compression stocking and intermittent pneumatic compression leg sleeves) and drug prophylaxis (unfractionated heparin and low molecular heparin), and the treatment is mainly thrombolysis or operative thrombectomy. The treatment of pulmonary artery embolism includes remittance of pulmonary arterial hypertension, anticoagulation, thrombolysis, operative thrombectomy, interventional therapy and extracorporeal membrane oxygenation (ECMO). Hemorrhage is a rarely occurred but relatively serious complication after bariatric surgery. The primary cause of anastomotic bleeding after laparoscopic gastric bypass is incomplete hemostasis or weak laparoscopic repair. The common bleeding site in laparoscopic sleeve gastrectomy is gastric stump and close to partes pylorica, and the bleeding may be induced by malformation and weak repair technique. Patients with hemodynamic instability caused by active bleeding or excessive bleeding should timely received surgical treatment. Anastomotic fistula in gastric bypass can be divided into gastrointestinal anastomotic fistula and jejunum-jejunum anastomotic fistula. The treatment of postoperative anastomotic fistula should vary with each individual, and conservative treatment or operative treatment should be adopted. Anastomotic stenosis is mainly related to the operative techniques. Stenosis after sleeve gastrectomy often occurs in gastric angle, and the treatment methods include balloon dilatation and stent implantation, and surgical treatment should be performed when necessary. Marginal ulcer after gastric bypass is a kind of peptic ulcer occurring close to small intestine mucosa in the junction point of stomach and jejunum. Ulcer will also occur in the vestige stomach after laparoscopic sleeve gastrectomy, and the occurrence site locates mostly in the gastric antrum incisal margin. Preoperative anti-HP (helicobacter pylorus) therapy and postoperative continuous administration of proton pump inhibitor (PPI) for six months is the main means to prevent and treat marginal ulcer. For patients on whom conservative treatment is invalid, endoscopic repair or surgical repair should be considered. Different surgical procedures will generate different related operative complications. Fully understanding and effectively dealing with the complications of various surgical procedures through multidisciplinary cooperation is a guarantee for successful operation.

Role of transient receptor potential vanilloid 1 in airway inflammation in asthmatic mice / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of the change in transient receptor potential vanilloid 1 (TRPV1) channel activity on the degree of airway inflammation in asthmatic mice.</p><p><b>METHODS</b>BALB/c mice were randomly divided into control, asthma, capsaicin (TRPV1 agonist), capsazepine (TRPV1 antagonist), and dexamethasone groups. The asthmatic mouse model was established by intraperitoneal injection of mixed ovalbumin-aluminium hydroxide solution and ultrasonic atomization with OVA for sensitization and challenge. The capsaicin, capsazepine, and dexamethasone groups were given intraperitoneal injection of capsaicin (30 μg/kg), capsazepine (10 μmol/kg), and dexamethasone (2 mg/kg) respectively, at 30 minutes before challenge. Hematoxylin and eosin staining was used to observe the degree of pulmonary inflammation. ELISA was used to measure the content of interleukin-8 (IL-8) and interleukin-13 (IL-13) in bronchoalveolar lavage fluid (BALF). Real-Time PCR was used to measure the relative content of TRPV1 mRNA in lung tissue.</p><p><b>RESULTS</b>Compared with the asthma group, the capsazepine and dexamethasone groups showed reduced pulmonary inflammation, while the capsaicin group showed aggravated pulmonary inflammation. Compared with the control group, the asthma and capsaicin groups showed increases in the content of IL-13 and IL-8 in BALF and the mRNA expression of TRPV1 in lung tissue (P<0.05). Compared with the asthma group, the capsazepine and dexamethasone groups showed reductions in the content of IL-13 and IL-8 in BALF and the mRNA expression of TRPV1 in lung tissue (P<0.05). The capsaicin group showed increases in the content of IL-13 and IL-8 in BALF (P<0.05).</p><p><b>CONCLUSIONS</b>TRPV1 channel agonist and antagonist can influence the degree of airway inflammation in asthmatic mice. Dexamethasone may reduce airway inflammation through regulating TRPV1 level.</p>