Exploring the molecular mechanism of Suoquan pill in the treatment of diabetic kidney disease based on network pharmacology,molecular docking,in vitro experiment

Background:Diabetic kidney disease(DKD)is a microvascular complication of diabetes mellitus and is the main cause of end-stage renal failure.Suoquan pills(SQP)has a variety of pharmacological activities and multiple therapeutic effects,and it is used clinically as a basic formula for the treatment of DKD.Methods:Public databases were used to identify SQP compounds and the potential targets of SQP and DKD.A drug-component-therapeutic target network was constructed.Protein-protein interaction network analysis,Gene Ontology functional analysis,and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to analyse the potential molecular mechanisms of SQP based on common targets of drugs and diseases.Molecular docking simulations were conducted to confirm the binding abity of the core compounds to key targets.The efficacy and predicted molecular mechanisms of SQP were validated using cell counting kit-8 assay,flow cytometry,and western blotting with HK-2 cells as a model.Results:Network pharmacology analysis showed that 26 compounds and 207 potential targets of SQP were involved in the treatment of DKD;boldine,denudatin B,pinocembrin,kaempferoid,and quercetin were considered core compounds,and epidermal growth factor receptor(EGFR)and proto-oncogene,non-receptor tyrosine kinase(SRC)were considered key targets.Gene Ontology enrichment analysis indicated that protein phosphorylation and negative regulation of apoptotic processes are important biological processes in the treatment of DKD by SQP.Molecular docking confirmed the excellent binding abilities of boldine,denudatin B,kaempferide,and quercetin to EGFR and SRC.The results of in vitro experiments showed that treatment with an ethanolic extract of SQP significantly protected HK-2 cells from high glucose-induced cell damage.In addition,the SQP ethanol extract inhibited the phosphorylation of EGFR and SRC,suppressed the apoptosis rate,and regulated apoptosis-related proteins in HK-2 cells under high glucose stress.Conclusion:This study systematically and intuitively illustrated the possible pharmacological mechanisms of SQP against DKD through multiple components,targets,and signalling pathways,especially the inhibition of EGFR and SRC phosphorylation and apoptosis.

Intravitreal slow-release dexamethasone alleviates traumatic proliferative vitreoretinopathy by inhibiting persistent inflammation and Müller cell gliosis in rabbits

AIM:To evaluate the effects of intravitreal slow-release dexamethasone on traumatic proliferative vitreoretinopathy(PVR)and Müller cell gliosis and preliminarily explored the possible inflammatory mechanism in a rabbit model induced by penetrating ocular trauma.METHODS:Traumatic PVR was induced in the right eyes of pigmented rabbits by performing an 8-mm circumferential scleral incision placed 2.5 mm behind the limbus,followed by treatment with a slow-release dexamethasone implant(Ozurdex)or sham injection.Left eyes were used as normal controls.The intraocular pressure(IOP)was monitored using an iCare tonometer.PVR severity was evaluated via anatomical and histopathological examinations every week for 6wk;specific inflammatory cytokine and proliferative marker levels were measured by quantitative real-time polymerase chain reaction,Western blot,protein chip analysis,or immunofluorescence staining.RESULTS:During the observation period,PVR severity gradually increased.Intense Müller cell gliosis was observed in the peripheral retina near the wound and in the whole retina of PVR group.Ozurdex significantly alleviated PVR development and Müller cell gliosis.Post-traumatic inflammation fluctuated and was persistent.The interleukin-1β(IL-1β)mRNA level was significantly upregulated,peaking on day 3 and increasing again on day 21 after injury.The expression of nod-like receptor family pyrin domain containing 3(NLRP3)showed a similar trend that began earlier than that of IL-1βexpression.Ozurdex suppressed the expression of IL-1β,NLRP3,and phosphorylated nuclear factor-kappa B(NF-κB).The average IOP after treatment was within normal limits.CONCLUSION:The present study demonstrates chronic and fluctuating inflammation in a traumatic PVR rabbit model over 6wk.Ozurdex treatment significantly inhibites inflammatory cytokines expression and Müller cell gliosis,and thus alleviates PVR severity.This study highlights the important role of IL-1β,and Ozurdex inhibites inflammation presumably via the NF-κB/NLRP3/IL-1βinflammatory axis.In summary,Ozurdex provides a potential therapeutic option for traumatic PVR.

Evaluation of the effect of pyrrolidine dithiocarbamate in suppressing inflammation in mice with dextran sodium sulfate-induced colitis

AIM: To evaluate the effect of pyrrolidine dithio- carbamate (PDTC; an NF-κB inhibitor) administered at low (50 mg/kg) and high (100 mg/kg) doses in suppressing colitis in mice with dextran sodium sulfate (DSS)-induced colitis. METHODS: Mice were divided into a DSS-untreated group (normal group), DSS-treated control group, DSS+PDTC-treated groupⅠ(low-dose group), and DSS+PDTC-treated groupⅡ (high-dose group). In each group, the disease activity index score (DAI score), intestinal length, histological score, and the levels of activated NF-κB and inflammatory cytokines (IL-1β and TNF-α) in tissue were measured. RESULTS: The DSS+PDTC-treated groupⅡ exhibited suppression of shortening of intestinal length and reduction of DAI score. Activated NF-κB level and IL-1β and TNF-α levels were significantly lower in DSS+PDTC- treated groupⅡ. CONCLUSION: These findings suggest that PDTC is useful for the treatment of ulcerative colitis.

Helicobacter pylori infection and inflammatory bowel disease in Asians:A meta-analysis

AIM: To investigate the relationship between Helicobacter pylori infection and inflammatory bowel disease(IBD) in an Asian population.METHODS: The Pub Med, EMBASE, and CochraneLibrary databases were searched for observational studies published up until June 2014, without language restrictions. Additional references were obtained from reviewed articles. RESULTS: Ten studies involving 1299 IBD patients and 1817 controls were included in the meta-analysis(24.9% of IBD patients had H. pylori infection vs 48.3% of the controls). The pooled risk ratio for H. pylori infection in IBD patients compared with controls was 0.48(95%CI: 0.43-0.54; P < 0.001). There was no significant heterogeneity in the included studies(I2= 21%). Egger's linear regression indicated that there was no significant publication bias(P = 0.203).CONCLUSION: The H. pylori infection rate in Asian IBD patients is significantly lower than in non-IBD patients, indicating that infection protects against the development of IBD.

Usefulness of fecal lactoferrin and hemoglobin in diagnosis of colorectal diseases

AIM:To evaluate prospectively usefulness of fecal lactoferrin(Lf)and fecal hemoglobin(Hb)in the diagnosis of colorectal diseases.METHODS:Fecal Lf and Hb were measured using ELISA in 872 patients before they underwent colorectal endoscopy.RESULTS:Lf was positive in 18(50%)of 36 patients with colorectal cancer,25(15.9%)of 157 with colorectal polyps,29(46.8%)of 62 with ulcerative colitis,and 25(62.5%)of 40(62.5%)with Crohn's disease.The Hb-positive rates were 50%,12.1%,41.9% and 32.5%,respectively.Of the 318 patients free of abnormalities by colorectal endoscopy,Lf was positive in 29(9.1%)and Hb was positive in 15(4.7%).Among patients with Crohn's disease,the Lf-positive rate was significantly higher than the Hb-positive rate.If either high Lf or Hb levels were considered positive,the positive rates rose to 61.1%,51.6%,and 67.5% in the colorectal cancer group,ulcerative colitis group,and Crohn's disease group,respectively.If both high Lf and Hb levels were rated positive,the positive predictive values(PPV)were 21% for colorectal cancer,33% for ulcerative colitis,and 17% for Crohn's disease,and PPV of high Hb level alone was 18%,25% and 13%,respectively.CONCLUSION:Fecal Lf and Hb were found useful in the detection of colorectal diseases,and the combination of the two measurements appears to increase the sensitivity and efficacy of diagnosis.

Characteristic expression of γ-aminobutyric acid and glutamate decarboxylase in rat jejunum and its relation to differentiation of epithelial cells

AIM: To investigate the expression between γ-aminobutyric acid (GABA) and glutamate decarboxylase and its relation with differentiation and maturation of jejunal epithelial cells in rat jejunum.METHODS: Immunohistochemical expression of GABA and glutamate decarboxylase (GAD, including two isoforms,GAD65 and GAD67) was investigated in rat jejunum.Meanwhile, double staining was performed with GAD65 immunohistochemistry, followed by lectin histochemistry of fluorescent wheat germ agglutinin. Furthermore,evaluation of cell kinetics in jejunum was conducted by ^3Hthymidine autoradiography and immunohistochemistry using a monoclonal antibody to proliferating cell nuclear antigen (PCNA).RESULTS: The cells showing positive immunoreactivity GABA and GAD65 were mainly distributed in the villi in rat jejunum, while jejunal epithelial cells were negative for GAD67. Positive GABA or GAD65 staining was mainly located in the cytoplasm and along the brush border of epithelial cells in the middle and upper portions. In addition, a few GABA and GAD65 strongly positive cells were scattered in the upper two thirds of jejunal villi. Double staining showed that GAD65 immunoreactivity was not found in goblet cells.^3H-thymidine-labeled nuclei were found in the lower and middle portions of jejunal crypts, which was consistent with PCNA staining. Therefore, GABA and GAD65 were expressed in a maturation or functional zone.CONCLUSION: The characteristic expression of GABA and GAD suggests that GABA might be involved in regulation of differentiation and maturation of epithelial cells in rat jejunum.

Confusing untypical intestinal Behcet's disease:Skip ulcers with severe lower gastrointestinal hemorrhage

Behcet's disease(BD) is a rare and life-long disorder characterized by inflammation of blood vessels throughout the body. BD was originally described in 1937 as a syndrome involving oral and genital ulceration in addition to ocular inflammation. Intestinal BD refers to colonic ulcerative lesions documented by objective measures in patients with BD. Many studies have shown that over 40% of BD patients have gastrointestinal complaints. Symptoms include abdominal pain, diarrhea, nausea, anorexia and abdominal distension. Although gastrointestinal symptoms are common, the demonstration of gastrointestinal ulcers is rare. This so-called intestinal BD accounts for approximately 1% of cases. There is no specific test for BD, and the diagnosis is based on clinical criteria. The manifestations of intestinal BD are similar to those of other colitis conditions such as Crohn's disease or intestinal tuberculosis, thus, it is challenging for gastroenterologists to accurately diagnose intestinal BD in patients with ileocolonic ulcers. However, giant ulcers distributed in the esophagus and ileocecal junction with gastrointestinal hemorrhage are rare in intestinal BD. Here, we present a case of untypical intestinal BD. The patient had recurrent aphthous ulceration of the oral mucosa, and esophageal and ileo-colonic ulceration, but no typical extra-intestinal symptoms. During examination, the patient had massive acute lower gastrointestinal bleeding. The patient underwent ileostomy after an emergency right hemicolectomy and partial ileectomy, and was subsequently diagnosed with incomplete-type intestinal BD by pathology. The literature on the evaluation and management of this condition is reviewed.

Unfavorable outcome of antiviral therapy in cytomegaloviruspositive ulcerative colitis may be due to inappropriate study inclusion in meta-analysis

Some previous articles reported that antiviral treatment was effective to reduce the colectomy rate in ulcerative colitis(UC) patients with cytomegalovirus(CMV) infection.Kopylov et al recently carried out a systematic review and meta-analysis to evaluate the impact of antiviral therapy on CMV-positive UC.The results showed that patients who received antiviral treatmenthad a higher risk of 30-d colectomy.We found that in this meta-analysis,some studies were inappropriately included,leading to an unfavorable outcome of antiCMV therapy in UC patients.

Histopathological and genetic differences between polypoid and non-polypoid submucosal colorectal carcinoma

AIM： To investigate the histopathological and geneticdifferences between polypoid growth （PG） and nonpolypoid growth （NPG） submucosal invasive colorectal carcinoma （CRC）.METHODS： A total of 96 cases of submucosal CRC were divided into two groups according to their growth type;60 cases of PG and 36 cases of NPG. The size, histological degree of dysplasia, depth of submucosal invasion and lymph node metastasis were compared between the two groups. Furthermore, expression of p53 was detected by immunohistochemical staining, and K-ras gene mutation was examined by polymerase chain reaction based single-strand conformation polymorphism （SSCP）.RESULTS： The average size of the lesions in the NPG group was significantly smaller than those in the PG group （7.5 mm vs 13.8 mm, P 〈 0.001）. The histological degree of dysplasia tended to be more severe in NPG group, while the incidence of submucosal massive invasion and the lymph node metastasis were both significantly higher in the NPG type than in the PG group （64.3% vs 43.3%, P = 0.004; 43% vs 7%, P =0.008, respectively）. In addition, K-ras gene mutations were detected in 67% of lesions in the PG group, but none in the NPG group, while no difference in p53immunohistochemical expression was found between the two groups.CONCLUSION： Compared with PG submucosal CRC,NPG type demonstrates more frequent submucosal massive invasion, more lymph node metastasis and a higher degree dysplasia. Genetically, NPG type shows much less frequent K-ras mutation.

Can admission lipoprotein-associated phospholipase A2 predict the symptomatic cerebral vasospasm following aneurysmal subarachnoid hemorrhage?

Background:Inflammation has been believed to be related to the development of cerebral vasospasm following aneurysmal subarachnoid hemorrhage(aSAH).A potential biomarker for vascular inflammation that is well recognized is the lipoprotein-associated phospholipase A2(Lp-PLA2).However,whether Lp-PLA2 can predict the occurrence of symptomatic cerebral vasospasm(SCV)in aSAH patients is still unknown.Thus,this study aimed to assess the value of Lp-PLA2 for predicting SCV in patients with aSAH.Methods:Between March 2017 and April 2018,we evaluated 128 consecutive aSAH patients who were admitted in the First Affiliated Hospital of Fujian Medical University.Their Lp-PLA2 level was obtained within 24 h of the initial bleeding.Factors might be related to SCV were analyzed.Results:Compared to patients without SCV,those with SCV(9.4%,12/128)had significantly higher Lp-PLA2 level.Multivariate logistic analysis revealed that worse modified Fisher grade(OR=10.08,95%CI=2.04–49.86,P=0.005)and higher Lp-PLA2 level(OR=6.66,95%CI=1.33–3.30,P=0.021)were significantly associated with SCV,even after adjustment for confounders.Based on the best threshold,Lp-PLA2 had a sensitivity of 83.3%and a specificity of 51.7%for predicting SCV,as shown by the receiver operating characteristic curve analysis.In the poor World Federation of Neurosurgical Societies grade patient sub-group,patients with Lp-PLA2>200μg/L had significantly higher SCV rate than that of patients having Lp-PLA2≤200μg/L.Conclusion:The admission Lp-PLA2 level might be a helpful predictor for SCV in aSAH.

Patient-specific ventricular puncture trajectory plane and puncture trajectory: a novel method of frontal ventricular puncture

Ventricular puncture is a common neurosurgical procedure,which is usually performed freehand using body surface anatomic landmarks as targeting references.[1,2]However,it is reported that the failure rate of freehand lateral ventricle puncture could be as high as 43%in traumatic brain injury cases if body surface anatomic landmarks are used as targeting references,[3]and the occurrence of hemorrhages associated with ventricular puncture could reach 16.2%.[4]Classic puncture sites,such as Kocher point,are helpful in improving the success rate of ventricular puncture.However,it is still risky to perform freehand puncture with classic puncture sites.Several causes could lead to this issue,including the following:(1)Individual patients have different conditions;a puncture approach suitable for all patients might not be possible.(2)Selection of puncture approach is largely dependent on the judgment of the surgeon,because the puncture approach does not usually fall on the coronal,sagittal,or axial images of the regular computed tomography(CT)scan;surgeons need to use spatial imagination based on CT images to design the ventricular puncture trajectory(VPT),which could lead to errors.(3)In freehand puncture,directional deviations could occur easily without the guidance of a frame as a fixed space reference to the puncture path.