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The present study investigated the therapeutic effect and mechanism of Di'ao Xinxuekang(DXXK) on non-alcoholic steatohepatitis(NASH) in mice. Sixty-five C57 BL/6 J mice were randomly divided into a normal group and an experimental group for model induction with the high-fat diet for 16 weeks. Then the mice in the experimental group were randomly divided into a model group, an atorvastatin group(4 mg·kg~(-1)·d~(-1)), and high-(200 mg·kg~(-1)·d~(-1)), medium-(60 mg·kg~(-1)·d~(-1)), and low-dose(20 mg·kg~(-1)·d~(-1)) DXXK groups, with 10 mice in each group. Drugs were administered by gavage for eight weeks. Serum lipid, liver lipid, serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione reductase(GSH-Px) were determined. Interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay(ELISA). The liver index was calculated. The liver pathological change and lipid accumulation were observed by HE and oil red O staining. The liver ultrastructure was observed by the transmission electron microscope. The mRNA and protein expression of nuclear factor-erythroid 2 related factor 2(Nrf2) and heme oxygenase-1(HO-1) was detected by real-time fluorescence-based quantitative PCR and Western blot, respectively. The results showed that compared with the normal group, the model group displayed serum lipid and liver lipid metabolism disorders, elevated transaminase, lipid deposition, steatosis, and inflammation, suggesting that the NASH model in mice was properly induced. Compared with the model group, the DXXK groups showed decreased serum lipid, liver lipid, ALT, AST, MDA, IL-1β, and TNF-α, increased SOD and GSH-Px, alleviated hepatic steatosis, ballooning, and inflammation, and up-regulated Nrf2 and HO-1 gene and protein expression. In conclusion, DXXK can significantly alleviate NASH in mice, which is related to the inhibition of oxidative stress and inflammatory damage by up-regulating the Nrf2/HO-1 signaling pathway.
Subject(s)
Animals , Mice , Drugs, Chinese Herbal , Inflammation/metabolism , Lipids , Liver , NF-E2-Related Factor 2/metabolism , Non-alcoholic Fatty Liver Disease/metabolism , Oxidative Stress , Signal Transduction , Superoxide Dismutase/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Nonalcoholic fatty liver disease （NAFLD） is a metabolic stress-induced liver injury characterized by excessive lipid accumulation in hepatocytes， which is closely related to insulin resistance and genetic susceptibility. It falls into the category of "liver lump" in traditional Chinese medicine （TCM）. NAFLD affects about 25% of the population worldwide and has become a major burden of the world health care system. However， its exact pathogenesis remains unclear. Conducting the basic research on NAFLD is of great clinical significance and social value. As an important tool for NAFLD research， animal model plays a particularly important role in clarifying the pathophysiological mechanism of NAFLD. In recent years， the modeling methods for NAFLD in China and abroad have been constantly updated， and in particular， certain progress has been made in the duplication of TCM syndrome models. By consulting and sorting out the relevant literature published in recent years in China and abroad， the author summarized the replication methods of NAFLD animal models. This paper reviewed the advantages and disadvantages of models established via dietary induction （high-fat feed， high-fat and high-fructose feed， high-fat and high-cholesterol feed， and methionine choline-deficient feed）， models with genetic defects ［leptin-deficiency （Lepob/Lepob）， autosomal recessive diabetes gene homozygous deficiency （ob/ob）， Alms1 gene （foz/foz） mutation， and FATZO mice］ and exposure to special diets， and models for TCM syndromes （liver depression and spleen deficiency syndrome， phlegm-dampness syndrome， blood stasis syndrome， combined phlegm and stasis syndrome， and qi stagnation and blood stasis syndrome）， in order to provide reference for the preparation of more scientific， reasonable， economical， and convenient animal models of NAFLD， thus laying a foundation for in-depth study of the pathogenesis， prevention， and treatment of NAFLD.
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Objective:To explore the effect and mechanism of Guizhitang with different proportions of Cinnamomi Ramulus and Paeoniae Alba Radix on overactive cardiac sympathetic nerves in salt-sensitive hypertensive rats. Method:Randomly divide the forty male 6-week-old salt-sensitive hypertensive rats into five groups： the normal control group， the model group， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶2 group，and the Cinnamomi Ramulus and Paeoniae Alba Radix 2∶1 group， each group has 8 animals， the normal control group was fed with low-salt feed， and the remaining four groups were fed with 8% high-salt feed. After 4 weeks of feeding， gastric feeding was started. Give both the normal control group and model group saline and the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group， the 1∶2 group，and the 2∶1 group， were given Guizhitang aqueous solution at 4.0， 5.5 and 5.5 g·kg-1， respectively. Continuous gavage intervention was held for 4 weeks. IITC multi-channel non-invasive sphygmomanometer was used to detect changes of systolic blood pressure before and after treatment in rats. Left ventricular anterior wall end-diastolic thickness （LVAWd） and interventricular septal diastolic thickness （IVSd） were detected by echocardiography. Hematoxylin-eosin（HE）staining and Masson staining were used to observe the myocardial morphological changes of rats in each group， Western blot was used to detect the expression of nerve growth factor （NGF）， growth-associated protein 43 （GAP43） and tyrosine hydroxylase （TH） protein. Result:After 4 weeks of intervention with Guizhitang， compared with the normal control group， the blood pressure， LVAWd and IVSd of the model group were significantly increased， and the expressions of NGF， TH and GAP43 protein were significantly increased （P<0.01）. HE and Masson staining results showed that the model group had myocardial cell edema， a large number of inflammatory cell infiltration， myocardial fiber hyperplasia and disordered arrangement， and a large amount of collagen deposition could be seen in the intercellular substance. Compared with model group， the systolic blood pressure of rats in each Guizhitang group increased slowly， and the expression of NGF， TH and GAP43 protein decreased （P<0.05，P<0.01），the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group has the best effect. The results of echocardiography shows that the 1∶1 Guishao group could reduce LVAWd and IVSd levels （P<0.05，P<0.01）， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶2 group could reduce IVSd level （P<0.05）， there was no statistical difference in LVAWd， there was no statistical difference in LVAWd and IVSd in 2∶1 group. In terms of myocardial morphology， each group of Guizhitang can reduce cell edema and inflammatory cell infiltration， reduce myocardial fiber hyperplasia and collagen deposition， and improve the disorder of myocardial fiber arrangement. Among them， the 1∶1 group has the best effect. Conclusion:Guizhitang can inhibit the overactive activation of cardiac sympathetic nervous system，reduce the extent of myocardial fibrosis， inflammatory infiltration and myocardial hypertrophy， and protect salt-sensitive hypertension rats， whose mechanism may be related to regulating the expression of heart NGF.Among them， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group is better than the 1∶2 and 2∶1 group in reducing myocardial fibrosis， inflammatory infiltration and myocardial hypertrophy.
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@#AIM:To investigate the expression of glucose key metabolic enzymes in human lens epithelial cells(HLEB3)induced by high glucose.<p>METHODS:HLEB3 cells cultured <i>in vitro</i> were divided into normal control group(DMEM medium containing 5mmol/L of glucose), oxidative stress group(DMEM medium containing 5mmol/L of glucose and 200μmol/L of hydrogen peroxide), high glucose induction group(DMEM medium containing 30mmol/L of glucose). Apoptosis and cells were detected 24h after culture. Activity and expression of six key glucose metabolic enzymes(fructose-6-phosphate kinase-1, pyruvate kinase, hexokinase, citrate synthase, α-ketoglutarate dehydrogenase and 6-phosphate glucose dehydrogenase)were studied.<p>RESULTS:Apoptosis of HLEB3 cells was induced by high glucose. The cell viability of high glucose-induced group(63.43%±3.40%)was lower than that of normal control group(100.00%±0.00%)and oxidative stress group(91.90%±5.11%), and the expression levels of six key enzymes of glucose metabolism in high glucose-induced group were lower than that of normal control group and oxidative stress group(all <i>P</i><0.05).<p>CONCLUSION:High glucose can induce the expression of glucose-key metabolizing enzymes in HLEB3 cells to decrease, induce apoptosis and affect cell activity.
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[Objective] To investigate the significance of liver biopsy in differential diagnosis and prognosis of congenital biliary atresia (CBA) and infant hepatitis syndrome (IHS).[Methods] Totally 77 children with congenital biliary atresia and 48 infants with hepatitis syndrome treated in Guangdong Women and Children Hospital from December 2012 to December 2016 were examined by liver biopsy and follow-up.Combined with immunohistochemistry and PAS staining,reticular fiber staining,Masson staining techniques,we make comparative analysis of both histopathological features and prognosis.[Results] The liver fibrosis grade,hepatic lobule inflammation activity staging,the degree of bile duct hyperplasia and the prognosis of CBA and IHS infants were statistically significant (P＜0.05).S2-S3-based liver fibrosis grading in infants with CBA,mainly in G2-G3 hepatic lobule inflammation staging,bile duct hyperplasia significantly;IHS infants with liver fibrosis grading as S0-S1,liver Slice inflammatory activity stage to G1-G2-based.The prognosis of infants with CBA was significantly worse than IHS,and the difference was statistically significant (P＜0.05).[Conclusion] The early liver biopsy of infants with congenital biliary atresia and infant hepatitis syndrome,combined with immunohistochemistry and PAS staining,reticular fiber staining,Masson staining techniques has important clinical significance to the differential diagnosis and prognosis of both.
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[Objective] To investigate the molecular mechanism of miR-192/-215 targeting BIVM in human gastric cancer.[Methods] First,BIVM was the target gene of miR-192/-215 screened by the target gene prediction in solico and gene microarrays.Real-time quantitative PCR verified the results of the microarrays.Then the double-luciferase reporter plasmids were constructed to test that BIVM was the target gene of miR-192/-215.Subsequently,BIVM-siRNA was transfected,to know the effects of BIVM-siRNA on proliferation and apoptosis of gastric cancer cells.8 nude mice were randomly divided into two groups:BIVM-siRNA experimental group and NSC-siRNA control group.Gastric cancer cells transfected with BIVM-SiRNA were implanted under the skin of nude mice to observe the effect of BIVM on the tumorigenicity of gastric cancer cells.[Results] BIVM was screened as miR-192/-215 target gene by gene microarrays and quantitative PCR.Double-luciferase reporting assays were performed to identify the BIVM as miR-192/-215 target gene.The cell proliferation assays showed that BIVM promoted the proliferation of gastric cancer cells (P＜0.05).Test of flow cytometry showed that BIVM inhibited the apoptosis of gastric cancer cells (P＜0.05).Mouse tumorigenesis test confirmed that BIVM could promote gastric cancer cells growth in vivo (P＜0.05).[Conclusion] BIVM plays a role in the carcinogenesis of gastric cancer,and miR-192/-215 targeting BIVM promotes the development of gastric cancer.
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The chemical constituents of Dioscorea opposite Thunb. were isolated and purified by Diaion HP-20, Sephadex LH-20, Toyopearl HW-40, MCI Gel CHP-20, ODS, silica gel column chromatography and semi-preparative-HPLC. Nine compounds were found and their structures were elucidated by spectral data and physicochemical properties, which were identified as 2-(1',2',3'-trihydroxybutyl-4'-O-α-D-glucopyranoside)-6-(2",3",4"-trihydroxybutyl)-pyrazine (1), uracil (2), xanthine (3), hypoxanthine (4), thymine (5), adenosine (6), uridine (7), inosine (8), and 5'-deoxy-5'-methylsulphinyladenosine (9). The compound 1 is a new pyrazine derivative, and the compound 3-5, 8, 9 are found in this plant for the first time.
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Objective: To establish the HPLC fingerprint and to determin gallic acid, methyl gallate, corilagin, and ellagic acid in Terminalia billerica, in order to provide the scientific foundation for quality control of T. billerica. Methods: The analysis was performed on Atlantic T3 (250 mm × 4.6 mm, 5 μm) C18 column, mobile phase was acetonitrile-0.2% glacial acetic acid aqueous solution with gradient elution, flow rate was 1.0 mL/min, injection size was 20 μL column, and temperature was maintained at 30 oC. The common mode of T. billerica HPLC fingerprint was established, the hidden information was analyzed in the fingerprint by Chemometrics, and the components in T. billerica by HPLC-MSn and quantitative analysis characteristic peaks were identified. Results: There were 21 common peaks in the diagram and the similarity of the fingerprints was over 0.9 in all 11 batches. The information of the 18 common peaks in T. billerica was summarized by HPLC-MSn technology. The samples were broadly divided into three kinds by principal component analysis and clustering analysis. The five key compounds were verified by partial least squares discriminant analysis method in quantitative analysis, and identified that the No.12 peak was chabulagic acid, and the average recoveries were in the range of 97.24%-98.58%. Conclusion: The HPLC fingerprint method and content determination method are reliable, accurate, rapid, simple, and reproducible, and this study could control the quality of T. billerica.
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Salvia miltiorrhiza is one of the most common traditional Chinese medicines. It has rich resources in China. According to modern studies, phenolic acids are the main effective components in S. miltiorrhiza. These components have cardiovascular and cerebrovascular protective effect, and anti-tumor, antioxidant, anti-inflammatory, and antifibrotic activities, etc. It has been widely used for the treatment of cardiovascular and cerebrovascular diseases and others. In this paper, the chemicals and pharmacological effects of phenolic acids from S. miltiorrhiza were summarized in the last decade. Its researches and development prospects were also analyzed for further studying and comprehensive utilization of these phenolic acids.
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<p><b>OBJECTIVE</b>To explore the clinicopathological characteristics of primary thyroid-like follicular carcinoma of the kidney.</p><p><b>METHODS</b>A case of primary thyroid-like follicular carcinoma of the kidney was studied with histology and immunohistochemical staining, and its clinical and pathological findings were further analyzed with review of the literature.</p><p><b>RESULTS</b>The patient was a 26-year-old asymptomatic woman who had a kidney mass during her annual physical examination. The tumor was well-circumscribed. Pathologically, the tumor showed follicular structures with colloid-like material in the lumina. Immunohistochemically, the tumor cells showed intense staining for CK7 and vimentin and negative for thyoid transcripation factor-1, thyroglobulin, thyoid peroxidase and RCC.</p><p><b>CONCLUSIONS</b>The diagnosis of primary thyroid-like follicular carcinoma of the kidney is based on the characteristic follicular architecture with colloid-like material, and the metastasis from a thyroid follicular carcinoma must be excluded clinically and pathologically before making the final diagnosis.</p>
Subject(s)
Adult , Female , Humans , Adenocarcinoma, Follicular , Metabolism , Pathology , DNA-Binding Proteins , Metabolism , Diagnosis, Differential , Follow-Up Studies , Keratin-7 , Metabolism , Kidney Neoplasms , Metabolism , Pathology , Nephrectomy , Methods , Neprilysin , Metabolism , Thyroid Neoplasms , Metabolism , Pathology , Transcription Factors , Vimentin , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the cause for allelic drop-out at short tandem repeat (STR) loci upon paternity testing with a PowerPlex® 16 kit.</p><p><b>METHODS</b>A total of 10 642 DNA confirmed paternity testing cases (18 314 parent/child allelic transfers) were analyzed with the PowerPlex® 16 kit. Samples suspected for having allelic drop-out were verified with an Identifiler™ kit and/or locus-specific singleplex amplification systems. PCR products of null alleles were separated and directly sequenced.</p><p><b>RESULTS</b>Eight cases of allelic drop-out were found. The overall rate of null allele in the PowerPlex® 16 system was 0.437 × 10(-3). DNA sequencing has confirmed single base variations within the binding region of published primers, in which 4 cases involved the D18S51 locus (2 cases with G>A transitions at 79 bp upstream of the repeats, 1 case with G>T transversion at 162 bp downstream of the repeats and 1 case with G>C transversion at 74 bp upstream of the repeats), 2 cases involved the D21S11 locus (1 case with C>A transversion at 17 bp upstream of the repeats and 1 case with A>G transition at 12 bp upstream of the repeats). One case involved the FGA locus (1 case with G>A transition at 142 bp downstream of the repeats) and 1 case involved TPOX locus (1 case with G>A transition at 198 bp downstream of the repeats).</p><p><b>CONCLUSION</b>Base variation in the primer binding region may cause failed PCR and result in null allele reports. Alternative primer sets should be used to verify the suspected allelic drop-out. Attention should be paid to this during paternity testing and data exchange for personal identification.</p>
Subject(s)
Humans , Male , Alleles , Genetic Variation , Microsatellite Repeats , Paternity , Sequence Analysis, DNAABSTRACT
<p><b>OBJECTIVE</b>To investigate the genetic polymorphisms of 12 X chromosome short tandem repeat (X-STR) loci of Investigator Argus X-12 amplification kit in Guangdong Han population.</p><p><b>METHODS</b>DNA samples from 200 unrelated individuals (100 males and 100 females) and 103 families (59 father-mother-daughter trios and 44 mother-son duos) were extracted and amplified with fluorescence labeled multiplex PCR system. PCR products were separated and genotyped with capillary array electrophoresis.</p><p><b>RESULTS</b>One hundred and thirty-seven alleles,including 9 off ladder alleles (OL allele) were observed at the 12 X-STR loci in the population. Six mutations were observed in 162 meioses. The combined power of discrimination (DP) was 0.999 999 997 in males and 0.999 999 999 in females, and the combined mean exclusion chance (MEC) was 0.999 999 988 in the trio cases and 0.999 998 013 in the duo cases.</p><p><b>CONCLUSION</b>Investigator-Argus X-12 amplification system is highly polymorphic in Guangdong Han population and it is powerful for personal identification and paternity testing.</p>
Subject(s)
Female , Humans , Male , Alleles , China , Chromosomes, Human, X , Gene Amplification , Gene Frequency , Genetics, Population , Genotype , Microsatellite Repeats , Mutation , Paternity , Polymerase Chain Reaction , Methods , Polymorphism, Genetic , RecordsABSTRACT
<p><b>OBJECTIVE</b>To investigate 16 to 22-year-old young adults' perception of malocclusion, dental health and aesthetic firstly. To investigate the perception differences between young adults and orthodontists, and factors on perception difference.</p><p><b>METHODS</b>A sample of 16 to 22-year-old young adults was selected randomly in Department of Stomatology, Hospital of Nanjing Army Command College. A questionnaire, which consisted of socioeconomic status, index of orthodontic treatment need and dental health component (DHC) questionnaire, was proposed to them. Index of orthodontic treatment need comprised aesthetic component (AC) and DHC. An orthodontist measured orthodontic dental models of participants. Participants chose AC degree and DHC degree by their subjective perception. An orthodontist chose AC degree and DHC degree by their objective measurement.</p><p><b>RESULTS</b>71.08% and 70.59% of 204 participants respectively chose 'no need treatment' of AC degree and DHC degree by their subjective perception. 19.12% and 9.80% of participants were respectively chosen 'no need treatment' of AC degree and DHC degree by orthodontist's objective measurement. Objective measurement degree of orthodontist was higher than subjective perception degree of participants. Subjective perception of participants was little related to objective measurement of orthodontists. AC degree of male participants' subjective perception was lower than the female. DHC degree of urban participants' subjective perception was higher than the rural. DHC and AC degree of only child participants' subjective perception was higher than non-only child.</p><p><b>CONCLUSION</b>Perception about malocclusion, dental health and aesthetic had significant difference between 16 to 22-year-old young adults and orthodontists. Young adults tended to think that their teeth were more beautiful and healthy. The female young adults than the male tended to think that their teeth were not enough beautiful. Urban young adults were more unsatisfactory about their dental health than the rural. Only child young adults were more unsatisfactory about their dental aesthetic and health than the non-only child.</p>
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Adolescent , Adult , Child , Female , Humans , Male , Young Adult , Esthetics, Dental , Health Services Needs and Demand , Malocclusion , Oral HealthABSTRACT
<p><b>OBJECTIVE</b>To compare the difference between J-hook and micro-implant anchorage in the treatment of patient with bimaxillary protrusion.</p><p><b>METHODS</b>Thirty patients with bimaxillary protrusion were divided into two groups (J-hook and micro-implant groups) and treated with MBT appliance. Four first premolars were extracted in all patients. Cephalometric analyses were carried out before and after treatment.</p><p><b>RESULTS</b>In J-hook group and micro-implant group,computerized cephalometric analysis revealed that before treatment U6C-PP was (12.4 +/- 0.2) mm and (12.5 +/- 0.1) mm, respectively,and after treatment U6C-PP was (12.6 +/- 0.1) mm and (12.8 +/- 0.1) mm,respectively. The difference between J-hook group and microimplant group was significant (P < 0.01). The other differences of cephalometric analyses between J-hook group and micro-implant group was not significant.</p><p><b>CONCLUSIONS</b>Both J-hook and micro-implant could provide adequate anchorage in the treatment of patients with bimaxillary protrusion.</p>
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Adolescent , Adult , Child , Female , Humans , Male , Young Adult , Malocclusion, Angle Class I , Therapeutics , Orthodontic Anchorage Procedures , Orthodontic Appliances , Orthodontics, Corrective , MethodsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the shape and structure of different separators affecting the mechanical behavior.</p><p><b>METHODS</b>The stress of different separators was analyzed by ANSYS software. Various separators were meshed into one-dimensional solid elements and the material character parameters were inputted. The loads were added gradually.</p><p><b>RESULTS</b>The stress was related to the arm of force and the point of force application.</p><p><b>CONCLUSIONS</b>The force applied can be controlled by selecting different separators.</p>