Effect of huankuile on colon injury in rats with ulcerative colitis by reducing TNF-α and MMP9.

OBJECTIVE: To explore the mechanism of huankuile (HKL) in colon injury repair in rats with ulcerative colitis (UC). METHODS: Fifty SPF Wistar male rats were divided randomly into a normal group, a negative control group, an HKL intervention group ('HKL group') and a 5-aminosalicylic acid intervention group ('5-ASA group'). After 14 days of intervention with corresponding drugs, pathological scores were obtained using the results of immunohistochemical staining; morphological changes were observed by hematoxylin-eosin staining, and the mRNA expression levels of tumour necrosis factor-α (TNF-α), matrix metalloproteinase 9 (MMP9) and interleukin-13 (IL-13) were detected by real-time quantitative PCR. RESULTS: After the successful construction of the rat model, it was compared with the rats in the normal group. In the negative group, it was found that the expression of TNF-α and MMP9 was significantly increased in the colonic mucosal epithelia of the rats, the pathological score was significantly increased (P < 0.05), and the mRNA expression levels of TNF-α, MMP9 and IL-13 were increased (P < 0.05). After treatment with HKL, the colonic morphology of the rats returned to normal, the expression of TNF-α and MMP9 in the colonic mucosal epithelium of the rats returned to normal, the pathological score grade was significantly reduced (P < 0.05), and the mRNA expression levels of TNF-α, MMP9 and IL-13 were reduced; these results were largely consistent with those of the normal group, with no statistically significant difference. CONCLUSION: HKL effectively improved the general symptoms and tissue injury in UC rats, and the therapeutic effect was better than that of 5-ASA group. Ulcerative colitis in rats increased the expression of TNF-α, MMP9 and IL-13. HKL repaired UC-induced colonic injury in rats by decreasing the expression of TNF-α, MMP9 and IL-13.

Lactobacillus acidophilus and HKL Suspension Alleviates Ulcerative Colitis in Rats by Regulating Gut Microbiota, Suppressing TLR9, and Promoting Metabolism.

Ulcerative colitis (UC) is a chronic non-specific inflammatory bowel disease with complex pathogenesis. The intestinal flora disturbance affects the homeostasis of the intestinal environment, leading to metabolic imbalance and immune abnormalities of the host, contributing to the perpetuation of intestinal inflammation. We suggest that the combination of anti-inflammatory therapy and the regulation of intestinal flora balance may help in the treatment process. Previously, we used a combination treatment consisting of Lactobacillus acidophilus (Lac) and Chinese medicine Huan Kui Le (HKL) suspension in a UC rat model, where the combined intervention was more effective than either treatment alone. Herein, the mechanism of action of this combined treatment has been investigated using 16S rRNA sequencing, immunohistochemistry, and ELISA methods in the colon, and untargeted metabolomics profiling in serum. Colon protein expression levels of IL-13 and TGF-ß were upregulated, whereas those of TLR9 and TLR4 were downregulated, consistent with an anti-inflammatory effect. In addition, gut microbiota structure changed, shown by a decrease in opportunistic pathogens correlated with intestinal inflammation, such as Klebsiella and Escherichia-Shigella, and an increase in beneficial bacteria such as Bifidobacterium. The latter correlated positively with IL-13 and TGF-ß and negatively with IFN-γ. Finally, this treatment alleviated the disruption of the metabolic profile observed in UC rats by increasing short-chain fatty acid (SCFA)-producing bacteria in the colonic epithelium. This combination treatment also affected the metabolism of lactic acid, creatine, and glycine and inhibited the growth of Klebsiella. Overall, we suggest that treatment combining probiotics and traditional Chinese medicine is a novel strategy beneficial in UC that acts by modulating gut microbiota and its metabolites, TLR9, and cytokines in different pathways.

The Antitumor Effect and Mechanism of Total Flavonoids From Coreopsis Tinctoria Nutt (Snow Chrysanthemum) on Lung Cancer Using Network Pharmacology and Molecular Docking.

Coreopsis tinctoria Nutt (C. tinctoria), also known as Snow Chrysanthemum, is rich in polyphenols and flavonoids. It has important pharmacological effects such as lowering blood lipids, regulating blood glucose, and anti-tumor effect. However, its anti-tumor mechanism has not yet been investigated thoroughly. This study aimed to explore the anti-tumor effect of total flavonoids extracted from C. tinctoria (CTFs) on lung cancer and the possible mechanism. The components of CTFs were analyzed using Ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The active components of CTFs were screened according to oral bioavailability (OB) and drug-likeness (DL). Totally, 68 components of CTFs were identified and 23 active components were screened. Network pharmacological analysis on the active components identified 288 potential targets associated with lung cancer. After protein-protein interaction (PPI) network topology analysis, 17 key protein targets including Akt1, MAPK1, TP53, Bcl-2, Caspase-3, Bax, GSK3B and CCND1 were screened. The molecular docking results showed that the active components of CTFs had good binding activity with key targets. GO and KEGG analysis of candidate targets found that the main enrichment was in PI3K/Akt-mediated intrinsic apoptotic pathways. Finally, according to the results of network pharmacology, the potential molecular mechanism of CTFs intervention in lung cancer was validated experimentally in vitro and in vivo. The experimental validation results demonstrated that the antitumor activity of CTFs on lung cancer may be related to inhibiting the PI3K-Akt signaling pathway and activating the mitochondrial-mediated apoptosis pathway.

Exploration of the second class activities in experimental teaching of medical biochemistry / 中华医学教育探索杂志

Experimental teaching is an indispensable task in teaching work in colleges and universities.Basic medical experimental teaching is the foundation of clinical practice,techniques,and scientific research.Medical biochemistry experiment is the compulsory experimental teaching content for medical students.The existing basic medical experimental teaching is far from enough for medical students.Therefore,it is very important to tap the potential of students,cultivate their interest in learning,improve the ability to learn on their own,operate manually,be creative,and solve problems,which can start from basic medical experimental teaching and be achieved through the second class activities.

Thyroid Dysfunction, Neurological Disorder and Immunosuppression as the Consequences of Long-term Combined Stress.

Stress is a powerful modulator of neuroendocrine, behavioral, and immunological functions. So far, the molecular mechanisms of response to stressors still remain elusive. In the current study, after 10 days of repeated chronic stress (hot-dry environment and electric foot-shock), a murine model of combined-stress (CS) was created in the SPF Wistar rats. Meanwhile, we established an ulcerative-colitis (UC) rat model induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS)/ethanol enema according to previous studies. The blood, hypothalamus, and colon tissues of these rats from CS, normal control (NC), UC and sham (SH) groups, were collected for further investigations. Comparing to the NC group, the serum levels of T3, T4, fT3 and fT4 were obviously decreased in the CS group after chronic stress, indicating that thyroid dysfunction was induced by long-term combined stress. Moreover, the application of RNA-seq and subsequent analyses revealed that neurological disorder and immunosuppression were also caused in the hypothalamus and colon tissues, respectively. Comparing with SH group, besides the induced colon inflammation, thyroid dysfuntion and neurological disorder were also produced in the UC group, suggesting that hypothalamic-pituitary-thyroid (HPT) axis and gastrointestinal system might not function in isolation, but rather, have intricate crosstalks.

Liposome encapsulated luteolin showed enhanced antitumor efficacy to colorectal carcinoma.

Luteolin is a falconoid compound that is present in various types of plants and possesses remarkable potential as a chemopreventive agent. However, the poor aqueous solubility of luteolin limits its clinical application. In the present study, an approach towards chemoprevention was explored using liposomes to deliver luteolin, and the antitumor efficacy was investigated in colorectal carcinoma. The present findings demonstrated that luteolin was efficiently encapsulated into liposomes with an encapsulation efficiency as high as 90%. The particle size of the liposomal luteolin (Lipo­Lut) and ζ­potential were optimized. In vitro studies demonstrated that, Lipo­Lut had a significant inhibitory effect on the growth on the CT26 colorectal carcinoma cell line compared with free luteolin (Free­Lut). The in vivo study indicated that Lipo­Lut could achieve superior antitumor effects against CT26 tumor compared with luteolin alone. The present results suggested that liposome delivery of luteolin improved solubility, bioavailability and may have potential applications in chemoprevention in clinical settings.

[Metabonomic analysis of the urine from rat model with abnormal sapra syndrome].

OBJECTIVE: To investigate the correlation between the change in metabolic components of urine and the abnormal sapra syndrome by using a rat model of abnormal sapra syndrome.
 Methods: Multiple factors, such as dry environment, dry feed, and chronic electrical stimulation, were used to establish the abnormal sapra syndrome in Wistar rats by Uyghur medicine. The differences in metabolites were detected through the metabonomics method.
 Results: The urine of rats in abnormal sapra syndrome group showed significant high abundance metabolites as follows: Leucine, isoleucine, and glycoprotein. And that significant low abundance metabolites as follows: Glutamine, creatine, citric acid, and phenylalanine.
 Conclusion: The urine of rats with the abnormal sapra syndrome displays abnormal energy metabolism. It is likely that the dysfunctional metabolisms of three major nutrients might be the molecular basis for the abnormal sapra syndrome.

Uygur medicine Xipayi Kui Jie'an affects gene expression profiles in intestinal tissue lesions in a rat model of ulcerative colitis.

BACKGROUND: The aim of this study was to investigate the mechanisms underlying the therapeutic effect of Uygur medicine KJA on UC in a rat model. METHODS: UC was induced in Wistar rats by application of 2, 4-dinitrochlorobenzene and acetic acid and were then treated with three different doses of KJA, and normal saline as control. After treatment for 20 days, the gene expression profile of colonic tissue was analyzed by microarray and verified by quantitative real-time RT-PCR. RESULTS: Animals treated with the three different doses of KJA were compared with normal saline controls, wherein microarray analysis identified 1991, 2163, and 1677 differentially expressed genes respectively, of which 444 genes were raised and 670 genes were decrease spliced together in the three doses tested. The KEGG pathway analyses found commonly raised genes related to several different biological functions. Interesting genes included TRL2, IL-1ß, TGF-ß1, and NF-κB were confirmed by quantitative PCR. CONCLUSIONS: The therapeutic effect of KJA on UC is likely explained by specific effects on the expression of genes, which are the effector molecules known to be involved in the development of UC. Further studies on differentially expressed genes will help explain the mechanism of action of Uygur medicine KJA.

[Metabonomic analysis of the serum from rat model with abnormal balgam syndrome of Uyghur medicine].

OBJECTIVE: To investigate correlation between the change in metabolic components of serum and the abnormal balgam syndrome by using a rat model of abnormal balgam syndrome. METHODS: Male Wistar rats were randomly divided into a control group and a test group. According to Uyghur medicine theory, the test group of rats were given wet cold diet (seeds of spinach and parsley, 24 hours) in a cold (6 °C) and humid (85%-95%, 10 hours) environment for 40 days to establish the rat model of abnormal balgam syndrome. 1H MR based metabonomic analysis of serum was performed. Data was analyzed using Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA) software. RESULTS: Compared with the control group, the serum components including glutamate, phenylalanine, tyrosine, citric acid, ß-hydrocxy butyrate, acetoacetate, pyruvic acid and creatine were decreased, while the glucose, lactic acid, low density lipoprotein and very low density lipoprotein were increased in the test group (P<0.05). CONCLUSION: The low energy production and consumption in the rat model of abnormal balgam syndrome suggests that the dysfunctional metabolisms of three major nutrients might be the molecular basis for the abnormal balgam syndrome.

Attempt of seminar in biochemistry teaching for Uygur clinical medicine undergraduate / 中华医学教育探索杂志

Objective To discuss the significance of the seminar in biochemistry teaching for Uygur clinical medical undergraduates. Methods 2011 grade totally 86 Uygur clinical medical under-graduates in Xinjiang Medical University were enrolled as the research object and questionnaire was used to make satisfaction survey. Data descript and analysis was performed according to the percentage of each option. Results 86.9%students thought that topic seminar was good;100.0%students thought that semi-nar can help apply and mastery the knowledge;100.0%students considered that seminar conducted dur-ing extracurricular time was necessary to the growth of knowledge;97.8%students considered that search-ing for references in the spare time was helpful;100.0%students were interested in the content of the seminar and thought it was novel and practical;100.0%students thought that seminar can help broaden thinking way;43.5%students supported that contents exceeding the syllabus can be used in examination while 39.1%students opposed; preparation time for examination lasted for 5 to 7 days. Conclusions Seminar applied in biochemistry teaching is very necessary,which can mobilize students' initiatives to learn actively and help consolidate and understand knowledge.

Construction of NF-kappaB siRNA expression vector / 中南大学学报(医学版)

OBJECTIVE@#To construct a NF-kappaB siRNA expression vector and to detect the specific silencing effect of the siRNA on the expression of NF-kappaB protein.@*METHODS@#pcDNA3.1/CT-GFP-TOPO recombinant eukaryotic expression vector and pSilencer 1.0-U6-siRNA-NF-kappaB recombinant vector were constructed respectively. These 2 recombination plasmids were co-transfected into COS-7 cells, and the NF-kappaB silence induced by RNAi was detected by Western blot and the inverted fluorescence microscope.@*RESULTS@#The levels of NF-kappaB protein in COS-7 cells could be silenced effectively and specifically by pSilencer 1.0-U6-siRNA- NF-kappa recombinant vector. The expression of NF-kappaB protein was reduced gradually with the increase of pSilencer 1.0-U6-siRNA- NF-kappaB recombinant vector,which could be detected by Western blot under the inverted fluorescence microscope.@*CONCLUSION@#NF-kappaB siRNA expression vector is constructed successfully.