Mechanism of Reversal of Precancerous Lesions of Gastric Cancer by Celastrus orbiculatus Extract via Regulating Lgr5/Wnt/β-catenin Signaling Pathway / 中国实验方剂学杂志

ObjectiveThis study aims to investigate the mechanism in which Celastrus orbiculatus extract （COE） affects the proliferation and differentiation of gastric organoids and the expression of Lgr5 and thus reverses the precancerous lesions of gastric cancer （PLGC） by regulating the leucine-rich repeat containing G protein-coupled receptor 5 （Lgr5）/Wingless （Wnt）/β-catenin signaling pathway based on a gastric organoid injury model. MethodGastric organoids were established based on stem cells of the mouse gastric gland. Gastric organoid injury models were constructed by treating gastric organoids with N-methyl-N'-nitro-N-nitrosoguanidine （MNNG， 0.02 mg·L-1）. Gastric organoid injury models were randomly divided into normal group， model group （0.02 mg·L-1 MNNG）， low， medium， and high dose （5， 10， 20 mg·L-1） groups of COE， and Wnt inhibitor Dickkopf-related protein 1 （DKK1） （0.5 mg·L-1） group， and they were treated with respective agents for 24 h. The number and volume of gastric organoids under different drug concentrations were observed under a microscope. The viability of the gastric organoid injury models was detected by Methyl thiazolyl tetrazolium （MTT） assay. The morphology and pathology of gastric organoids were observed using Hematoxylin and Eosin （HE） staining. The expression levels of Lgr5， Mucin2 （MUC2）， Mucin5AC （MUC5AC）， Mucin6 （MUC6）， Wnt， and β-catenin in gastric organoids under different drug concentrations were detected by Western blot （WB）. ResultCompared with the normal group， the number， volume， and activity of gastric organoids in the model group were decreased （P<0.01）， while the expressions of Lgr5， MUC2， Wnt， and β-catenin were significantly increased （P<0.01）. The expressions of MUC5AC and MUC6 were significantly decreased （P<0.01）. Compared with the model group， the number and volume of gastric organoids in the low， medium， and high dose groups of COE were all improved （P<0.01）， and the vitality of gastric organoids was significantly enhanced （P<0.01）. The effect was the most significant at a COE concentration of 20 mg·L-1 （P<0.01）. The expressions of Lgr5 and MUC2 in the medium and high dose groups of COE were significantly reduced （P<0.01）， while the expression of MUC5AC and MUC6 were significantly increased in the low， medium， and high dose groups of COE （P<0.05， P<0.01）. Compared with the model group， Wnt inhibitors could promote the expression of MUC5AC and MUC6 in gastric organoids （P<0.05， P<0.01） and reduce the expression of MUC2， Wnt， and β-catenin. In addition， the combined use of COE at high concentrations and Wnt inhibitors could further promote this trend （P<0.01）. ConclusionCOE inhibits the Wnt/β-catenin pathway by inhibiting the expression of Lgr5， MUC2， Wnt， and β-catenin and promoting the expression of MUC5AC and MUC6， thus promoting the proliferation and differentiation of gastric organoids and reversing the PLGC process.

Correlation between the Expression of Serum Elabela,LRG1 Levels and Disease Activity Index in Patients with Ulcerative Colitis / 现代检验医学杂志

Objective To investigate the expression of serum Elabela and leucine-rich-alpha-2-glycoprotein-1(LRG1)in ulcerative colitis(UC)patients and their correlation with disease activity index(DAI).Methods A total of 98 patients with UC admitted to Yuncheng Central Hospital from January to December 2022 were selected as the UC group,including 62 patients in active stage and 36 patients in remission stage.According to the severity of the disease,these patients were divided into mild group(n=26),moderate group(n=43)and severe group(n=29).In addition,these patients were grouped into gradeⅠ group(n=25),grade Ⅱ group(n=40)and grade Ⅲ group(n=33)based on the endoscopic activity index(EAI).According to the mucosal healing condition under endoscopy,these patients were divided into the healed group(n=65)and the unhealed group(n=33).Another 51 patients with colonic polyps were selected as control group 1,and 50 healthy individuals were selected as control group 2.Serum Elabela and LRG1 levels were detected by enzyme-linked immunosorbent assay(ELISA).Pearson method was used to analyze the correlation between serum Elabela,LRG1 levels and DAI in UC patients.Receiver operating characteristic(ROC)curve was applied to analyze the predictive value of serum Elabela and LRG1 for endoscopic mucosal healing.Results The levels of Elabela(4.77±1.36 ng/ml)and LRG1(352.12±39.45 ng/ml)in UC group were higher than those in control group 1(2.51±0.53 ng/ml,121.02±21.06 ng/ml)and control group 2(2.35±0.42 ng/ml,120.35±23.49 ng/ml),and the differences were statistically significant(t= 11.410～39.000,all P＜0.05).The levels of Elabela(5.26±0.54 ng/ml)and LRG1(370.42±12.49 ng/ml)in the active group were higher than those in the remission group(3.93±0.42 ng/ml,320.60±8.47 ng/ml),and the differences were statistically significant(t=12.705,21.242,all P＜0.05).The levels of Elabela(5.89±0.20 ng/ml)and LRG1(369.92±16.59 ng/ml)in the severe group were higher than those in the moderate groups(4.51±0.67 ng/ml,356.12±18.75 ng/ml)and mild groups(3.95±0.21 ng/ml,325.65±10.14 ng/ml),and the differences were statistically significant(t=3.205～35.077,all P＜0.05).The levels of Elabela(5.80±0.18 ng/ml)and LRG1(369.16±13.47 ng/ml)in grade Ⅲ group were higher than those in grade Ⅱ group(4.49±0.35 ng/ml,355.46±16.34 ng/ml)and grade Ⅰgroup(3.86±0.16 ng/ml,324.15±8.71 ng/ml),and the differences were statistically significant(t= 3.854～48.725,all P＜0.05).The levels of Elabela(5.12±0.42 ng/ml)and LRG1(367.12±14.27 ng/ml)in unhealed group were higher than those in healed group(4.08±0.37 ng/ml,322.57±10.35 ng/ml),and the differences were statistically significant(t=12.043,15.917,all P＜0.05).The serum levels of Elabela and LRG1 in UC patients were positively correlated with EAI and ESR(r=0.602,0.298;0.576,0.302,all P＜0.05),but negatively correlated with hemoglobin level(r=-0.351,-0.334,all P＜0.05).The area under the curve predicted by the combination of serum Elabela and LRG1 for endoscopic mucosal healing was 0.926(95%CI:0.880～0.958),was higher than the 0.803(95%CI:0.741～0.856)and 0.783(95%CI:0.720～0.838)predicted by Elabela and LRG1 alone,and the difference was statistically significant(Z=4.101,4.228,all P＜0.05).Conclusion The serum levels of Elabela and LRG1 in UC patients increased,and they were related to the increase of DAI and worsening of the condition.Testing serum Elabela and LRG1 can provide a reference for evaluating mucosal healing under UC endoscopy.

Discussion on the Targeting Mechanism of Qingre Huoxue Formula （清热活血方） for Treatment of Rheumatoid Arthritis by Regulating “Inflammation-Bone Destruction”：Based on LRG1/TGF- β Signalling Axis / 中医杂志

Based on the previous publications， it is believed that damp-heat syndrome is the core syndrome of rheumatoid arthritis （RA）， and Qingre Huoxue Formula （清热活血方） is an effective formula for the treatment of damp-heat syndrome of RA. “Inflammation-bone destruction” is a key pathological link of RA， and it is also the advantage of the effectiveness of Qingre Huoxue Formula. Leucine rich α-2-glycoprotein 1 （LRG1） can mediate the transforming growth factor-β （TGF-β） signalling pathway to participate in the pathogenic process of “inflammation-bone destruction” of RA， and it can be used as a target protein in the treatment of damp-heat syndrome of RA by Qingre Huoxue Formula. Accordingly， a scientific hypothesis was proposed that Qingre Huoxue Formula may regulate TGF-β signalling pathway mediated by LRG1 to improve “inflammation-bone destruction” of RA， and it was envisioned that the clinical effect of Qingre Huoxue Formula on LRG1 could be confirmed through clinical studies， and the mechanism of action of Qingre Huoxue Formula on the LRG1/TGF-β signalling axis as well as the influence of the expression or non-expression of the LRG1/TGF-β signalling axis on the therapeutic effectiveness of Qingre Huoxue Formula could be clarified through animal experiments.

Methylene blue alleviates dopaminergic neuronal pyroptosis to improve motor dysfunction in Parkinson's disease mouse models / 中华神经医学杂志

Objective:To investigate the effect of methylene blue (MB) on motor dysfunction and its mechanism in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) mouse models.Methods:Forty healthy male C57BL/6 mice were randomly divided into 4 groups: control group, model group, low-dose treatment group and medium-dose treatment group ( n=10); PD mouse models were established by intraperitoneal injection of 25 mg/kg/d MPTP for a consecutive 7 d; low-dose treatment group and medium-dose treatment group were pretreated intraperitoneally with MB 2 mg/kg/d or MB 10 mg/kg/d for a consecutive 3 d, respectively; and then, MPTP 25 mg/kg/d+MB 2 mg/kg/d or MPTP 25 mg/kg/d+MB 10 mg/kg/d were injected intraperitoneally into the low-dose treatment group or medium-dose treatment group for a consecutive 7 d (MPTP and MB were given at 12 h of interval). Eight d after modeling, open field experiment, pole climbing experiment and rod rotating experiment were carried out to evaluate the spontaneous movement, coordination, endurance and motor ability. And then, the mice were sacrificed; immunofluorescent staining was used to observe tyrosine hydroxylase (TH) expression in the substantia nigra; Western blotting was used to detect the expressions of TH, α-synuclein, nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), cleaved-Caspase-1 and Gasdermin D (GSDMD) in the striatum and substantia nigra of mice. Contents of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-18 in the substantia nigra and striatum of mice were detected by ELISA. Results:Compared with the control group, the model group had shortened residence time in rod rotating, prolonged descent time in rod climbing, reduced total movement distance in open field, decreased number of TH-positive cells in the substania nigra, decreased TH protein levels in the substania nigra and striatum, and increased NLRP3, ASC, cleaved-Caspase-1, GSDMD and GSDMD-N protein levels in the substania nigra and striatum, and increased TNF-α, IL-1β and IL-18 contents in the substania nigra and striatum, with significant differences ( P<0.05). Compared with the model group, low-dose treatment group and medium-dose treatment group had prolonged residence time in rod rotating, shortened descent time in rod climbing, increased total movement distance in open field, increased number of TH-positive cells in the substania nigra, and increased TH protein levels in the substania nigra and striatum, decreased NLRP3, ASC, and cleaved-Caspase-1 levels in the substania nigra and striatum, and decreased TNF-α, IL-1β and IL-18 contents in the substania nigra and striatum, with significant differences ( P<0.05). No statistical differences in the above indexes were noted between the low-dose treatment group and medium-dose treatment group ( P>0.05). Conclusion:Low-/medium-dose MB can ameliorate motor dysfunction in PD mouse models, whose mechanism may be related to downregulate NLRP3 inflammasome and inhibit neuroinflammatory response to reduce dopaminergic neuron pyroptosis.

Role of post-translational modification of basic leucine zipper transcription factors in response to abiotic stresses in plants / 生物工程学报

Abiotic stresses substantially affect the growth and development of plants. Plants have evolved multiple strategies to cope with the environmental stresses, among which transcription factors play an important role in regulating the tolerance to abiotic stresses. Basic leucine zipper transcription factors (bZIP) are one of the largest gene families. The stability and activity of bZIP transcription factors could be regulated by different post-translational modifications (PTMs) in response to various intracellular or extracellular stresses. This paper introduces the structural feature and classification of bZIP transcription factors, followed by summarizing the PTMs of bZIP transcription factors, such as phosphorylation, ubiquitination and small ubiquitin-like modifier (SUMO) modification, in response to abiotic stresses. In addition, future perspectives were prospected, which may facilitate cultivating excellent stress-resistant crop varieties by regulating the PTMs of bZIP transcription factors.

Causal relationship between branched-chain amino acids and peripheral atherosclerosis： a Mendelian randomization study / 上海预防医学

ObjectiveWe conducted a two-sample Mendelian randomization （MR） study to assess the causal relationship between circulating isoleucine， leucine and valine levels and the risk of peripheral atherosclerosis. MethodsBased on the large-scale genome-wide association study （GWAS） database， single nucleotide polymorphisms （SNPs） closely related to the circulating levels of isoleucine， leucine and valine were identified as instrumental variables （IVs）. Two-sample MR analysis applying the inverse variance weighted （IVW） method and the weighted median estimator （WME） method were performed to estimate the causal relationship between the risk of peripheral atherosclerosis and the exposure with more than three SNPs that were available as IVs. The pleiotropy was evaluated by using the MR-Egger regression and MR-PRESSO method， and the leave-one-out method was used in sensitivity analysis. ResultsFour， one and one SNPs were identified as IVs for circulating isoleucine， leucine and valine levels， respectively. For isoleucine， the IVW model demonstrated there was no evidence of heterogeneity among the IVs （P=0.715）， and there was a significant causal relationship between the increase of circulating isoleucine level and a higher risk of peripheral atherosclerosis risk. Per every 1 elevated standard deviation （SD） of circulating isoleucine level resulted in increasing 31% of peripheral atherosclerosis risk （OR=1.31， 95%CI： 1.07‒1.61）. Similarly， the OR（95%CI） was 1.33 （1.04‒1.71） in the WME model. The MR-Egger regression and MR-PRESSO analysis indicated no evidence of pleiotropy in IVs （all P>0.05）. The result of the leave-one-out sensitivity analysis was stable. The Wald ratio model displayed that the causal relationship between circulating leucine and valine levels and the risk of peripheral atherosclerosis was not statistically significant. The OR （95%CI） for leucine and valine was 1.13 （0.78‒1.63） and 1.11 （0.82‒1.50）， respectively. ConclusionThere is a significant causal relationship between the increase of circulating isoleucine level and a higher peripheral atherosclerosis risk. The causal relationships between circulating leucine and valine levels and the risk of peripheral atherosclerosis need to be further confirmed in future studies.

LRG1 inhibits hepatic macrophage activation by enhancing TGF-β1 signaling to alleviate MAFLD in mice / 南方医科大学学报

OBJECTIVE@#To explore the effect of leucine-rich α-2-glycoprotein (LRG1) derived from hepatocytes on activation of hepatic M1 Kupffer cells.@*METHODS@#A metabolic dysfunction-associated fatty liver disease (MAFLD) model was established in BALB/c mice by high-fat diet (HFD) feeding for 16 weeks. Oleic acid was used to induce steatosis in primary cultures of mouse hepatocytes. The mRNA and protein expressions of LRG1 in mouse liver tissues and hepatocytes were detected by real-time PCR and Western blotting. Primary hepatic macrophages were stimulated with the conditioned medium (CM) from steatotic hepatocyte along with LRG1 or transforming growth factor-β1 (TGF-β1), or both for 24 h, and the expression levels of inducible nitric oxide synthase (iNOS) was detected with Western botting, and the mRNA expressions of iNOS, chemokine ligand 1 (CXCL-1) and interleukin-1β (IL-1β) were measured by RT-PCR. The MAFLD mice were injected with LRG1 (n=6), TGF-β1 (n=6), or both (n=6) through the caudal vein, and the live tissues were collected for HE staining and immumohistochemical detection of F4/80 expression; the mRNA expressions of iNOS, CXCL-1 and IL-1β in liver tissues were detected using RT-PCR.@*RESULTS@#The mRNA and protein expression levels of LRG1 were significantly downregulated in the liver tissues of MAFLD mice and steatotic hepatocytes (P < 0.05). Treatment of the hepatic macrophages with CM from steatosis hepatocytes significantly enhanced the mRNA expression levels of iNOS, CXCL-1 and IL-1β, and these changes were significantly inhibited by the combined treatment with TGF-β1 and LRG1 (P < 0.05). In MAFLD mice, injections with either LRG1 or TGF-β1 alone reduced hepatic lipid deposition and intrahepatic macrophage infiltration, and these effects were significantly enhanced by their combined treatment, which also more strongly inhibited the mRNA expression levels of iNOS, CXCL-1 and IL-1β (P < 0.05).@*CONCLUSION@#LRG1 inhibits hepatic macrophage infiltration by enhancing TGF-β1 signaling to alleviate fatty liver inflammation in MAFLD mice.

Transcription factor EB attenuates neuronal damage following cerebral ischemia via regulating autophagy-lysosomal pathway / 国际脑血管病杂志

The disorder of autophagy lysosomal pathway (ALP) is an important pathogenesis of neuronal damage after cerebral ischemia, and the restoration of ALP may alleviate neuronal damage after cerebral ischemia. As the main transcription factor regulating ALP, transcription factor EB (TFEB) can directly regulate autophagosome generation, autophagosome-lysosome fusion, and autophagic flux by regulating the expression of autophagic genes and lysosomal genes. Therefore, regulating TFEB can alleviate ALP dysfunction and thereby reduce cerebral ischemic damage. This article reviews the structure, biological function of TFEB and its role in regulating ALP to alleviate neuronal damage after cerebral ischemia.

Correlation of serum antibody titers with clinical features and prognosis of anti leucine-rich glioma inactivated 1 antibody encephalitis / 中华神经科杂志

Objective:To analyze the relationship between serum antibody titers, clinical characteristics, and prognosis in patients with encephalitis induced by anti leucine-rich glioma inactivated 1 (LGI1) antibody.Methods:Clinical data of 20 patients diagnosed with encephalitis in the Department of Neurology, First Hospital of Shanxi Medical University from February 2015 to February 2021 were collected and retrospectively analyzed. Patients were divided into 2 groups based on their serum anti LGI1 antibody titers, namely the high titer group (1∶100, 1∶320) and the low titer group (1∶10, 1∶32). The clinical characteristics, laboratory test results, and prognosis of the 2 groups of patients were compared. Relusts The age of the 20 patients with anti LGI1 antibody encephalitis ranged from 27 to 69 (53.5±11.2) years, with a male to female ratio of 1∶1. There were 9 patients in the low titer group and 11 patients in the high titer group. There was no statistically significant difference in the types and quantities of clinical symptoms between the 2 groups. Patients in the high titer group were more prone to abnormal lesions on cranial magnetic resonance imaging (10/11 vs 3/9, P=0.014). There was no statistically significant difference between the 2 groups in the presence or absence of cerebrospinal fluid anti LGI1 antibodies (9/11 vs 4/9, P=0.160). During the follow-up, it was found that 1/20 patient died of pulmonary embolism, 7/20 of patients were able to recover to their predisease state, and 9/20 of patients had residual memory impairment. In the high titer group, 3/11 of patients experienced recurrence, while there was no recurrence in the low titer group. There was no statistically significant difference in the neurological prognosis between the 2 groups at 3 months of discharge and follow-up (the number of patients whose modified Rankin Scale score≤2: 10/10 vs 8/9, P=0.474). Conclusions:Patients with high serum anti LGI1 antibody titers are more likely to develop intracerebral lesions. Higher antibody titers may be associated with a higher risk of disease recurrence. There was no significant correlation between serum antibody titers and neurological outcomes.

Expression and clinical significance of maternal embryo leucine zipper kinase in renal clear cell carcinoma / 国际生物医学工程杂志

Objective:To investigate the expression of maternal embryonic leucine zipper kinase (MELK) in renal clear cell carcinoma and its relationship with clinical and pathological characteristics of the patients with renal clear cell carcinoma, and to evaluate its effect on the proliferation of renal clear cell carcinoma.Methods:The expression of MELK in renal clear cell carcinoma and its relationship with the overall survival of patients was analyzed using the gene expression profiling interactive analysis (GEPIA) online database. Cancer tissue samples from 77 patients with renal clear cell carcinoma treated at Xinxiang Central Hospital were collected, all of which have been confirmed as renal clear cell carcinoma through immunohistochemistry experiments. The relationship between the expression of MELK in renal clear cell carcinoma tissue and the clinical and pathological characteristics of the patients was analyzed. The effect of MELK on the proliferation of renal clear cell carcinoma cells was explored through cloning experiments and CCK-8 assay.Results:Information from the GEPIA database revealed that the expression level of the MELK gene was upregulated in renal clear cell carcinoma compared with normal tissues. The clinicopathological analysis results showed that MELK expression was significantly correlated with tumor size and diameter of renal clear cell carcinoma ( P < 0.05), and had no statistically significant correlation with patient age, gender, or tumor differentiation. The results of the clonogenic assay and the CCK-8 assay showed that MELK expression promoted the proliferation of renal clear cell carcinoma cells (all P < 0.05). Conclusions:MELK promotes the proliferation of kidney cancer cells, which may provide a new therapeutic strategy for the treatment of renal clear cell carcinoma.

Clinical characteristics of 31 patients with anti-leucine-rich glioma inactivated-1 antibody-associated encephalitis / 临床神经病学杂志

Objective To improve the clinical recognition of anti-leucine-rich glioma inactivated-1(LGI1)antibody associated encephalitis through investigating the clinical characteristics of the disease,so as to improve diagnosis and prognosis.Methods A total of 31 consecutive patients with anti-LGI1 antibody associated encephalitis were recruited during January 2020 to March 2022 in Brain Hospital Affiliated to Nanjing Medical University including their clinical manifestations.Patients were divided into first-line immunotherapy group and mycophenolate mofetil(MMF)adding-on group according to the initial treatment regimen.Univariate and multivariate COX regression analysis were used to analyze the factors of the recurrence of anti-LGI1 antibody encephalitis.mRS was used to evaluate the prognosis.Results Seizure(93.5%),memory defect(80.6%)and psychosis(58.1%)were the most common manifestations in 31 patients.Fourteen patients suffered with hyponatremia,1 patients with elevated white cells and 17 patients with elevated proteins in CSF.The corresponding positive rates of anti-LGI1 antibody were 90.3%and 93.1%in serum and CSF.Fourteen patients had abnormal cranial MR.All patients received first-line immunotherapy,with 13 patients followed by MMF.Eleven patients were recurred in the follow-up.Univariate COX regression analysis found female(HR =3.85,95%CI:1.12-13.24,P =0.032),MMF adding-on therapy(HR = 4.03,95%CI:1.07-15.22,P = 0.04)were associated with relapse.However no independent associations were found after multivariate COX regression analysis.Twenty-two patients had sequeleas during follow-up,with memory decline as the most common symptom.All patients achieved good prognosis.Conclusions Anti-LGI1 antibody associated encephalitis may have good prognosis,but relapse is not rare.The majority patients have sequelaes,with memory defect and psychosis being the most common symptoms.MMF might have no prevention effect on anti-LGI1 antibody encephalitis recurrence.

Prokaryotic expression and polyclonal antibody preparation of LRRC15 protein, an excretory-secretory antigen of Cysticercus cellulosae / 中华地方病学杂志

Objective:To construct a recombinant plasmid pET30a-leucine-rich repeat (LRR) containing 15 (LRRC15) of Taenia solium, prokaryotically express and purify the LRRC15 recombinant protein, and prepare a rabbit polyclonal antibody. Methods:The LRRC15 protein encoding gene of Taenia solium was obtained by whole gene synthesis; it was cloned into pET30a vector, and the recombinant plasmid pET30a-LRRC15 was constructed and identified by double-enzyme PCR; the recombinant plasmid was transformed into competent cells of Escherichia coli BL21 (DE3), and the recombinant protein LRRC15 was induced to express by isopropyl-beta-D-thiogalactopyranoside (IPTG), the expression product was analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); the LRRC15 recombinant protein was purified by Ni-IDA affinity columns, the purified recombinant protein was analyzed and identified by SDS-PAGE, and the specificity of the purified recombinant protein was identified by Western blot (WB); the New Zealand rabbits were immunized with purified LRRC15 recombinant protein to prepare polyclonal antibodies against LRRC15, and the potency of the purified polyclonal antibody was determined by indirect enzyme-linked immunosorbent assay (ELISA). Results:After PCR identification, a band with a length of 1 506 bp was amplified, which was consistent with the LRRC15 gene; after SDS-PAGE and WB identification, the LRRC15 target protein with a relative molecular mass ( Mr) of about 55.36 × 10 3 was obtained; after immunizing New Zealand rabbits with purified LRRC15 recombinant protein, a polyclonal antibody against LRRC15 was obtained, and its potency was 1∶1 587 200. Conclusion:The recombinant plasmid pET30a-LRRC15 is successfully constructed, the LRRC15 recombinant protein of Taenia solium is prepared, and a high purity and high potency rabbit anti polyclonal antibody against LRRC15 recombinant protein is obtained.

Correlation of serum and follicular fluid SREBP1c and LRG1 levels with insulin resistance in PCOS patients / 中华内分泌代谢杂志

Objective:To investigate the serum and follicular fluid levels of sterol regulatory element-binding protein 1c(SREBP-1c), leucine-rich α-2-glycoprotein 1(LRG1) and the correlation with insulin resistance(IR) in non-ovarian etiology infertility patients and polycystic ovary syndrome(PCOS) patients with or without IR.Methods:Forty-nine PCOS patients and 66 infertility patients with non-ovarian etiology were collected in this retrospective study, homeostasis model assessment for insulin resistance(HOMA-IR) was used to evaluate IR, and were divided into control group( n=36), IR group( n=30), PCOS alone group( n=28) and PCOS-IR group(PCOS with IR group, n=21). The concentrations of serum, follicular fluid LRG1 and SREBP1c levels in each group were compared, and their correlation with relevant hormones and glycolipid metabolism were analyzed. Results:The levels of serum, follicular fluid LRG1 and SREBP1c in IR group, PCOS alone group and PCOS-IR group were significantly higher than those in control group; The PCOS-IR group showed a more significant increase in the levels of serum, follicular fluid LRG1 and SREBP1c( P<0.05). Correlation analysis showed that serum, follicular fluid LRG1 was positively correlated with body mass index, fasting plasma glucose(FPG), fasting insulin(FINS), triglycerides(TG), and HOMA-IR( P<0.05). Serum, follicular fluid SREBP1c was positively correlated with body mass index, FPG, FINS, TG, total cholesterol, LDL-C, LH, total teststerone, DHEAS, FAI, and HOMA-IR( P<0.05). Binary logistic regression analysis showed that serum SREBP1c was a risk factor for PCOS( P<0.05). Conclusion:The serum and follicular fluid levels of LRG1 and SREBP-1c were elevated in PCOS patients, especially in those with IR. The elevated levels of serum and follicular fluid LRG1 and SREBP-1c may be associated with IR and glucose-lipid metabolism abnormalities in PCOS patients. Serum LRG1 and SREBP-1c levels may serve as new indicators for predicting IR, early diagnosis, and intervention in PCOS patients.

Expression of LRG-1 in mice with hypertensive renal damage and its significance / 中南大学学报(医学版)

OBJECTIVES@#Long-term elevated blood pressure may lead to kidney damage, yet the pathogenesis of hypertensive kidney damage is still unclear. This study aims to explore the role and significance of leucine-rich alpha-2-glycoprotein-1 (LRG-1) in hypertensive renal damage through detecting the levels of LRG-1 in the serum and kidney of mice with hypertensive renal damage and its relationship with related indexes.@*METHODS@#C57BL/6 mice were used in this study and randomly divided into a control group, an angiotensin II (Ang II) group, and an Ang II+irbesartan group. The control group was gavaged with physiological saline. The Ang II group was pumped subcutaneously at a rate of 1.5 mg/(kg·d) for 28 days to establish the hypertensive renal damage model in mice, and then gavaged with equivalent physiological saline. The Ang II+irbesartan group used the same method to establish the hypertensive renal damage model, and then was gavaged with irbesartan. Immunohistochemistry and Western blotting were used to detect the expression of LRG-1 and fibrosis-related indicators (collagen I and fibronectin) in renal tissues. ELISA was used to evaluate the level of serum LRG-1 and inflammatory cytokines in mice. The urinary protein-creatinine ratio and renal function were determined, and correlation analysis was conducted.@*RESULTS@#Compared with the control group, the levels of serum LRG-1, the expression of LRG-1 protein, collagen I, and fibronectin in kidney in the Ang II group were increased (all P<0.01). After treating with irbesartan, renal damage of hypertensive mice was alleviated, while the levels of LRG-1 in serum and kidney were decreased, and the expression of collagen I and fibronectin was down-regulated (all P<0.01). Correlation analysis showed that the level of serum LRG-1 was positively correlated with urinary protein-creatinine ratio, blood urea nitrogen, and blood creatinine level in hypertensive kidney damage mice. Serum level of LRG-1 was also positively correlated with serum inflammatory factors including TNF-α, IL-1β, and IL-6.@*CONCLUSIONS@#Hypertensive renal damage mice display elevated expression of LRG-1 in serum and kidney, and irbesartan can reduce the expression of LRG-1 while alleviating renal damage. The level of serum LRG-1 is positively correlated with the degree of hypertensive renal damage, suggesting that it may participate in the occurrence and development of hypertensive renal damage.

Efectos de la proteína whey, leucina y entrenamiento de fuerza en la composición corporal / Effects of whey protein, leucine and resistance training on body composition

INTRODUCCIÓN: Los deportistas y las personas activas que entrenan la fuerza para lograr un aumento de su masa muscular tienen requerimientos calóricos y proteicos aumentados, por esta razón emplean diversas estrategias como la suplementación con proteína whey aislada y aminoácidos aislados como la leucina después del entrenamiento sustentados en la premisa de que promueven el incremento en la síntesis de proteínas musculares, y por ende un aumento de la masa magra. OBJETIVO: Comparar cambios de la composición corporal aplicando el método de Absorciometría Dual de Rayos X (DEXA) en personas físicamente activas expuestas a un programa de entrenamiento de la fuerza con la suplementación de proteína whey aislada y/o leucina durante 12 semanas. METODOLOGÍA: estudio aleatorizado prospectivo longitudinal en un grupo de 10 varones físicamente activos, los cuales fueron asignados al azar en 3 grupos de suplementación con proteína whey aislada, leucina o la mezcla de ambas bajo el mismo protocolo de entrenamiento de la fuerza. Se evaluó la composición corporal en 3 momentos: semana 0, semana 6 y semana 12 de la intervención. RESULTADOS: En las condiciones evaluadas no se observaron cambios estadísticamente significativos de las variables de composición corporal y fuerza en los diferentes protocolos de suplementación: leucina, proteína whey aislada con leucina y proteína whey aislada. CONCLUSIONES: Se observaron resultados superiores de la masa magra y la fuerza en el grupo de la suplementación de proteína whey aislada con leucina con respecto a los demás grupos, cabe aclarar que por el tamaño de muestra no alcanzó a ser estadísticamente significativo.

INTRODUCTION: Athletes and active people who work hard to increase their muscle mass have increased caloric and protein requirements. For this reason, they use different strategies such as whey protein isolate supplementation and/or branched-chain amino acids after sustained training on the premise that they promote an increase in muscle protein synthesis, and therefore an increase in lean mass. OBJECTIVE: To compare body composition changes in physically active people exposed to a strength training program withwhey protein isolate and/or leucine supplementation for 12 weeks. METHODS: Longitudinal prospective randomized study in a group of 10 physically active males, which were randomly assigned to 3 supplementation groups with whey protein isolate, leucine, or a mixture of both, under the same strength training protocol. Body composition was evaluated in 3 moments: week 0, week 6 and week 12 of the intervention. RESULTS: In the evaluated conditions, no statistically significant changes were observed in the variables of body composition and strength based on the different supplementation protocols: Leucine, protein with leucine and protein. CONCLUSIONS: Compared with the other groups, higher results were observed in lean mass and strength in group using the whey protein isolate supplementation with leucine. It should be noted that due to the size of the sample, the difference was not statistically significant.

Expression of BZW2 and IVD in hepatocellular carcinoma tissues and effect on prognosis of liver transplant recipients with hepatocellular carcinoma / 器官移植

Objective To investigate the expression levels of basic leucine zipper and W2 domain 2 (BZW2) and isovaleryl-CoA dehydrogenase (IVD) in hepatocellular carcinoma (HCC) and evaluate their effect on clinical prognosis of liver transplant recipients with HCC. Methods Pathological specimens and clinical data of 87 liver transplant recipients with HCC were collected and retrospectively analyzed. The recurrence and metastasis of HCC after liver transplantation were assessed. Immunohistochemical staining was used to detect the expression levels of BZW2 and IVD. The relationship between BZW2, IVD and clinicopathological parameters of HCC and their effect on postoperative recurrence and clinical prognosis of the recipients was analyzed. Results Among 87 recipients, 31 cases recurred with a recurrence rate of 36%. HCC recurred at postoperative 2-49 months and the median recurrence time was postoperative 7 months. Immunohistochemical staining demonstrated that the positive expression rate of BZW2 in the HCC tissues was significantly higher than that in normal liver tissues (76% vs. 30%), and the positive expression rate of IVD was significantly lower compared with that in normal liver tissues (51% vs. 69%) (both P < 0.01). BZW2 expression was significantly correlated with tumor diameter and tumor capsule (both P < 0.05), whereas IVD expression was significantly associated with tumor diameter, alpha-fetoprotein (AFP) level, tumor, node and metastasis (TNM) staging and whether vascular invasion was found or not (all P < 0.05). In the high BZW2 expression group, the cumulative recurrence rate of HCC was significantly higher and the cumulative survival rate was significantly lower than those in the low BZW2 expression group. In the low IVD expression group, the cumulative recurrence rate of HCC was significantly higher and the cumulative survival rate was significantly lower compared with those in the high IVD expression group (all P < 0.05). Conclusions The expression level of BZW2 protein is up-regulated, whereas that of IVD protein is down-regulated in the HCC tissues. Moreover, the cumulative recurrence rate of HCC is relatively high and the cumulative survival rate is relatively low in liver transplant recipients with high BZW2 expression and low IVD expression.

Effects of small leucine-rich proteoglycans on corneal transparency / 中华实验眼科杂志

Small leucine-rich proteoglycans (SLRPs) are necessary structural ingredients of the cornea, which are vital for the establishment and maintenance of corneal transparency.SLRPs are mainly located in the corneal stroma and can be divided into class Ⅰ, class Ⅱ, and class Ⅲ.The compensatory and cooperative interactions among SLRPs regulate the formation and assembly of stromal collagen fibrils, thereby maintaining the highly ordered arrangement of collagen fibrils, and establishing corneal transparency.Decorin and lumican are the main functional components of class Ⅰ and class Ⅱ SLRPs, respectively, and changes in their expression or abnormities in the structure of their core proteins affect the natural content and arrangement of other stromal extracellular matrix components, ultimately resulting in abnormal fibril formation, assembly, and arrangement, causing corneal opacity.SLRPs can regulate corneal wound healing and stromal matrix remodeling via binding to fibrotic molecules and their receptors, which provides bases for corneal diseases therapy and study of molecular mechanisms of corneal transparency.The bioactivities and the role of SLRPs in corneal transparency were reviewed in this article.

The expression of leucine zipper EF-hand domain transmembrane protein 1, sodium hydrogen exchange protein 1 and adenylate cyclase related protein 2 in gastric cancer tissues and their predictive value for postoperative recurrence / 中国医师进修杂志

Objective:To investigate the expression of leucine zipper EF-hand domain transmembrane protein 1 (LETM1), sodium hydrogen exchange protein 1 (NHE-1) and adenylate cyclase related protein 2 (CAP2) in gastric cancer tissues and the predictive value for postoperative recurrence.Methods:The clinical data of 92 patients with early gastric cancer who underwent surgical treatment from January 2017 to January 2020 in Jiangsu Haimen People′s Hospital were retrospectively analyzed. According to the recurrence condition 6 months after operation, the patients were divided into recurrence group (16 cases) and non recurrence group (76 cases). The expression levels of LETM1, NHE-1 and CAP2 mRNA in cancer tissues and adjacent tissues were detected by real time fluorescent quantitative polymerase chain reaction. Multifactor Logistic regression analysis was used to analyze the related influencing factors of postoperative recurrence. The receiver operating characteristic (ROC) curve was drawn, and the effectiveness of LETM1, NHE-1 and CAP2 mRNA in predicting recurrence was analyzed. The Kaplan-Meier survival curve was drawn, and the survival rate was analyzed in patients with different expression levels of LETM1, NHE-1 and CAP2 mRNA.Results:The LETM1, NHE-1 and CAP2 mRNA in cancer tissues were significantly higher than those in adjacent tissues (0.41±0.12 vs. 0.22±0.07, 0.85±0.27 vs. 0.49±0.15 and 0.31±0.10 vs. 0.19±0.06), and there were statistical differences ( P<0.01). The proportion of N 1 stage in recurrent group was significantly higher than that in non recurrent group: 9/16 vs. 22.37% (17/76), and there was statistical difference ( P<0.05); the LETM1, NHE-1 and CAP2 mRNA of cancer tissues in recurrent group were significantly higher than those in non recurrent group (0.61±0.20 vs. 0.37±0.13, 1.24±0.38 vs. 0.77±0.21 and 0.60±0.19 vs. 0.25±0.10), and there were statistical differences ( P<0.01). Multifactor Logistic regression analysis result showed that, after controlled N stages, the LETM1, NHE-1 and CAP2 mRNA were still independent risk factors for postoperative recurrence in patients with gastric cancer ( OR = 1.52, 3.11 and 1.21; 95% CI 1.26 to 1.82, 2.36 to 4.09 and 1.04 to 1.41; P<0.01). ROC curve analysis result showed that the area under the curve (AUC) of LETM1, NHE-1 and CAP2 mRNA for predicting postoperative recurrence in patients with gastric cancer were 0.768, 0.802 and 0.850, respectively. The AUC of combination the indexes for predicting postoperative recurrence in patients with gastric cancer was 0.965. According to the cut-off value of ROC curve, the patients were divided into LETM1 mRNA high expression (>0.54, 30 cases) and low expression (≤0.54, 62 cases), NHE-1 mRNA high expression (>1.09, 35 cases) and low expression (≤1.09, 57 cases), CAP2 mRNA high expression (>0.49, 28 cases) and low expression (≤0.49, 64 cases). Kaplan-Meier survival curve analysis result showed that the survival rates in patients with high expression of LETM1, NHE-1 and CAP2 mRNA were significantly lower than those in patients with low expression (73.33% vs. 98.39%, 80.00% vs. 96.49% and 78.57% vs. 95.31%), and there were statistical differences ( χ2 = 15.08, 6.95 and 6.75, P<0.01). Conclusions:LETM1, NHE-1 and CAP2 mRNA are related to the recurrence of early gastric cancer after surgery. The detection of the 3 markers is expected to provide a new strategy for the prediction of postoperative recurrence.

Morvan syndrome with positive anti LGI1/CASPR2 antibodies in serum/cerebrospinal fluid:a case report and literature review / 中华内科杂志

To report a typical case of Morvan syndrome with positive anti-leucine rich glioma-inactivated 1(LGI1) and contactin-associated protein 2 (CASPR2) antibodies in serum and cerebrospinal fluid. A 39-years-old female initially presented weakness of extremeties. The main symptoms included paroxysmal limb pain, wheezing, itching, muscle twitching, epilepsy, hypomnesia, dysphoria, apathy, intractable insomnia, salivation and sweating. Tests of electrolytes found hypokalemia (2.7-3.1 mmol/L) and hyponatremia (130-136 mmol/L). Arterial blood gas analysis showed hypoxemia (oxygen saturation 50%-70%). Total thyroxine (TT4) was elevated to 207 nmol/L with positive thyroid peroxidase antibody (TPO-Ab) and thyroglobulin antibody (TG-Ab). LGI1and CASPR2 antibodies (CBA method) were positive in both serum and cerebrospinal fluid, and the remaining antibodies related to autoimmune encephalitis and paraneoplastic syndrome were negative. Head MRI was almost normal, while mild abnormalities were found in electroencephalogram. Electromyography showed slightly increased voltage of left quadriceps motor unit potential. After treated with corticosteroids, IVIG and mycophenolate mofetil, the patient completely improved. Cognitive function scores recovered from MoCA/MMSE (16/24) to MoCA/MMSE (26/29). Positivity of LGI1/CASPR2 antibodies both in serum/cerebrospinal fluid are rarely seen in patients with Morvan syndrome. Steroids and immunosuppressants are suggested for treatment as early as possible.

Expression and promoter methylation status of leucine rich repeat containing 55 in pancreatic carcinoma tissue and the clinical value / 中华胰腺病杂志

Objective:To detect the mRNA expression and methylation status of leucine rich repeat containing 55(LRRC55) gene in pancreatic carcinoma tissues, and discuss the clinical value.Methods:Resected pancreatic ductal adenocarcinoma and normal adjacent specimens from 37 patients admitted in General Surgery of First Affiliated Hospital of Naval Medical University were collected from May 2019 to May 2021. Another two normal pancreas specimens and two blood samples from healthy adults were also collected. All patients′ age, gender, tumor location, tumor size, tumor differentiation, TNM staging, lymphatic metastasis, CEA and CA19-9 level were recorded. Bisulfite treatment of genomic DNA and sequencing analysis was used to study methylation patterns in CpG islands of the promoter for LRRC55 gene in fresh tissues from 2 pancreatic adenocarcinoma and adjacent tissues, 2 normal pancreatic tissues, 2 pancreatic cancer cell lines (PaTu8988 and ASPC1). LRRC55 mRNA in 35 pancreatic adenocarcinoma and adjacent tissues was detected by real-time quantitative PCR and the correlations with clinical parameters were analyzed.Results:CpG islands of LRRC55 in pancreatic adenocarcinoma tissues and pancreatic cancer cell lines was highly methylated and the mean methylation rate was 53% and 71%, respectively; while LRRC55 gene in pancreatic adjacent tissues and normal pancreatic tissues was lowly methylated, and the mean methylation rate was 8% and 11%. The relative expression in the pancreatic adenocarcinoma tissues and the paired adjacent normal tissues was 0.21 (0.02, 1.00 ) and 0.98 (0.33, 3.66 ), respectively; the former was significantly lower than the later and the difference was statistically significant ( P=0.003). Correlation analysis showed that LRRC55 mRNA expression level was related to tumor differentiation and CEA, but not correlated with patients′ age, gender, tumor location and size, CA19-9 level, lymphatic metastasis and TNM staging. Conclusions:Pancreatic cancer tissue and cell lines had abnormal methylation of LRRC55 gene; LRRC55 gene hypermethylation was related with its lower mRNA expression level in pancreatic cancer, which was correlated with the tumor differentiation and CEA level. LRRC55 may be a potential suppressor gene for pancreatic cancer.