OBJECTIVES: To investigate the effect of a variety of kidney-tonifying Chinese medicines on thymus regeneration after acute degeneration in mice. METHODS: Forty-eight 8-week-old male BALB/c mice were randomly divided into normal control group, model control group, negative control group, positive control group, the fructus of Cnidium monnieri (L.) Cuss. group, the fructus of Psoralea corylifolia (L.) group, the fructus of Rubus chingii Hu group, and the tuber onion seed group, with 6 mice in each group. Except for the normal control group, mice in the other groups received intraperitoneal injections of rapamycin (1 mg·kg^－1·d^－1) for 5 consecutive days followed by 14 h of starvation to induce acute thymus degeneration. After successful modeling, in treatment groups ethanol extract of the fructus of Cnidium monnieri (L.) Cuss. (0.78 g·kg^－1·d^－1), fructus of Psoralea corylifolia (L.) (0.39 g·kg^－1·d^－1), fructus of Rubus chingii Hu (0.78 g·kg^－1·d^－1) or the tuber onion seed(0.39 g·kg^－1·d^－1) was intraperitoneally injected once a day for 5 days; while the negative control group was given equal volume of normal saline, and the positive control group was given metformin (300 mg·kg^－1·d^－1). The grip strength was measured with a grip tester 2 h after the last administration. The pathological changes of thymus were observed by hematoxylin and eosin (HE) staining. The structure and distribution of thymic epithelial cells were observed by multiple immunofluorescence method. The proportion of T cell subsets in thymus and peripheral blood was analyzed by flow cytometry. The level of T cell receptor excision circles (TREC) in the genomic DNA of mouse spleen mononuclear cells was detected by quantitative polymerase chain reaction (PCR) for evaluation of thymic output function. The expression of thymus aging- and function-related factors in the thymus tissue were detected by quantitative reverse transcription PCR. The expression of cyclin-dependent kinase inhibitor 1A (p21) and tumor protein 53 (p53) were verified by immunohistochemistry. RESULTS: Rapamycin induced thymic atrophy and significantly reduced limb grip strength in mice (P<0.01). Compared with the negative control group, the limb grip strength of mice in the fructus of Psoralea corylifolia (L.) group, the fructus of Rubus chingii Hu group and the tuber onion seed group was significantly enhanced (all P<0.05), and the level of TREC in spleen of the mice in each administration group was reduced (all P<0.05). Among Chinese herb medicine-treatment groups, the recovery of thymus function and tissue structure in the tuber onion seed group was most obvious. Further study showed that compared with the negative control group, the proportion of CD4 single positive cells (CD3⁺TCR-β⁺CD4⁺CD8^－) in the thymus of the tuber onion seed group was significantly increased (P<0.01), and the proportion of CD3⁺CD28⁺ T cell and CD3⁺CD8⁺CD28⁺ T cell in peripheral blood was significantly increased (all P<0.01). The mRNA levels of IL-1α, IL-6, p21 and p53 in thymocytes were decreased (all P<0.05). The results of immunohistochemistry further confirmed the decrease in p21 and p53 expression. In normal mice, tuber onion seed was observed to enhance limb grip strength (P<0.01), while suppressing thymus output and change the distribution of T cell subsets, and there was no significant effect on thymus weight and the expression of Foxn1, SIRT1, p21, CXCL2 and PTMα. CONCLUSIONS The tuber onion seed and other kidney-tonifying traditional Chinese medicines can accelerate the regeneration process of mouse thymus after acute degeneration induced by rapamycin in mice, and the tuber onion seed exhibits the most pronounced therapeutic effect.
Animals ; Mice ; Male ; Mice, Inbred BALB C ; Thymus Gland/physiology* ; Drugs, Chinese Herbal/pharmacology* ; Sirolimus/adverse effects* ; Regeneration/drug effects*

ObjectiveTo investigate the therapeutic effect of ganoderic acid A （GA-A） on a mouse model of concanavalin A （ConA）-induced autoimmune hepatitis （AIH）. MethodsA total of 35 mice were randomly divided into control group （NC group）， model group （ConA group）， and low-， middle-， and high-dose GA-A treatment groups （GA-A-L， GA-A-M， and GA-A-H groups， respectively）， with 7 mice in each group. ConA was injected via the caudal vein of mice to establish a classic mouse model of AIH， and different doses of GA-A were administered via intraperitoneal injection 1 hour later for treatment. Proteomic techniques were used to investigate the protective mechanism of GA-A on hepatocytes， and HL-60 cells were differentiated into dHL-60 neutrophils by all-trans retinoic acid in vitro to validate the mechanism of action of GA-A. Related indicators were measured， including inflammatory markers （the activities of serum alanine aminotransferase ［ALT］ and aspartate aminotransferase ［AST］， HE staining， and inflammation-related genes）， apoptosis markers （TUNEL staining）， neutrophils， and neutrophil extracellular trap （NET） markers （myeloperoxidase ［MPO］， citrullinated histone H3 ［CitH3］， Ly6G， and free double-stranded DNA ［dsDNA］）， and p38 phosphorylation markers. The independent samples t-test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the NC group， the ConA group had significant increases in the serum levels of ALT and AST （both P<0.001）， and compared with the ConA group， GA-A treatment significantly reduced the levels of ALT and AST （both P<0.01）. HE staining showed that the mice in the ConA group had significant liver necrosis， while GA-A treatment significantly reduced the area of liver necrosis and the number of TUNEL-positive cells （both P<0.05）. Compared with the ConA group， the GA-A group had significant reductions in the expression levels of the inflammatory factors interleukin-6， tumor necrosis factor-α， and interferon gamma in serum and liver tissue （all P<0.05）. The proteomic analysis showed that GA-A alleviated ConA-induced acute immune liver injury by inhibiting the release of NET and the p38 MAPK pathway. Immunofluorescent staining of mouse liver tissue showed that compared with the ConA group， the GA-A group had significant reductions in the number of MPO-positive neutrophils and the number of cells with positive Ly6G and CitH3 （all P<0.01）. Western Blot and dsDNA testing showed that GA-A significantly inhibited the levels of the NET markers dsDNA and CitH3 and the level of p38 phosphorylation in liver tissue and dHL-60 cells （all P<0.05）. ConclusionGA-A alleviates liver inflammatory response and hepatocyte death by inhibiting the p38 MAPK pathway and the release of NET， thereby alleviating ConA-induced acute immune liver injury. This study provides a theoretical basis for the use of GA-A to treat immune liver injury by regulating neutrophil function.

Objective:To determine lysophosphatidic acid receptor 6 (LPAR6) expression in patients with mycosis fungoides (MF) , a variant of cutaneous T-cell lymphoma (CTCL) , and to investigate its role and mechanism of action in the development and prognosis of CTCL.Methods:A total of 110 patients with confirmed MF were collected from Department of Dermatology, Peking University First Hospital from 2011 to 2020, including 24 with large-cell transformation (LCT) and 25 with non-large cell transformation (NLCT) in the discovery cohort, and 24 with LCT and 37 with NLCT in the validation cohort. RNA sequencing and RT-PCR were conducted to determine the LPAR6 expression in patients in the discovery cohort and validation cohort respectively. LPAR6 expression was compared between patients with LCT and those with NLCT, and its effect on the prognosis of patients was evaluated. Two LPAR6-overexpressing CTCL cell lines MyLa and Sz4 were constructed to evaluate the effect of LPAR6 overexpression on proliferative activity of MyLa and Sz4 cells, with the cells normally expressing LPAR6 as the control group; after the treatment with LPAR6-related ligand lysophosphatidic acid (LPA) , 2S-OMPT, adenosine triphosphate (ATP) or adenosine (ADO) , the effects of LPAR6 activation on the proliferative activity and apoptosis of LPAR6-overexpressing MyLa and Sz4 cells were evaluated by the MTS method and flow cytometry respectively. Log-rank test was used for prognostic analysis, and t test or Mann-Whitney U test was used for comparisons between two groups. Results:As RNA sequencing showed, LPAR6 was one of the significantly underexpressed genes in the LCT group in the discovery cohort; in the validation cohort, LPAR6 expression (median[ Q1, Q3]) was significantly lower in the LCT group (204.90[81.90, 512.70]) than in the NLCT group (809.40[417.50, 1 829.20], U= 242.00, P= 0.002) ; in the two cohorts, the underexpression of LPAR6 was significantly associated with increased risk of poor prognosis (both P < 0.01) . Cell proliferation assay showed no significant difference in the proliferative activity of MyLa or Sz4 cells between the LPAR6 overexpression group and control group at 0, 24, 48 and 72 hours during the experiment (all P > 0.05) ; 48 hours after activation of LPAR6 by LPA, 2S-OMPT, ATP and ADO in MyLa cells, the LPAR6 overexpression group showed significantly decreased cellular proliferative activity (1.38 ± 0.01, 1.04 ± 0.01, 1.09 ± 0.03, 1.23 ± 0.01, respectively) compared the control group (1.73 ± 0.04, 1.23 ± 0.01, 1.24 ± 0.01, 1.42 ± 0.03, t= 30.33, 18.38, 4.78, 5.75, respectively, all P < 0.05) , but significantly increased cell apoptosis rate (17.93% ± 0.88%, 17.75% ± 0.35%, 23.97% ± 0.57%, 31.44% ± 0.34%, respectively) compared the control group (3.98% ± 0.03%, 7.81% ± 0.59%, 11.95% ± 0.85%, 12.02% ± 0.48%, t= 15.93, 14.49, 11.74, 33.01, respectively, all P < 0.05) ; 48 hours after activation of LPAR6 by 2S-OMPT and ADO in Sz4 cells, compared with the control group, the LPAR6 overexpression group also showed significantly decreased cellular proliferative activity (2S-OMPT: 1.29 ± 0.04 vs. 1.48 ± 0.01; ADO: 1.27 ± 0.01 vs. 1.51 ± 0.02; both P < 0.05) , but significantly increased cell apoptosis rate (2S-OMPT: 41.70% ± 0.70% vs. 29.35% ± 0.55%; ADO: 37.05% ± 0.15% vs. 24.60% ± 1.00%; both P < 0.05) . Conclusions:LPAR6 was underexpressed in the patients with LCT, and its underexpression was significantly associated with increased risk of poor prognosis. In vitro activation of LPAR6 could inhibit the proliferation of CTCL cells and promote their apoptosis, suggesting that the decrease of LPAR6 expression may be one of the important mechanisms underlying disease progression in patients with LCT.

Object:This study aims to compare the efficacy, acceptability and tolerability of cognitive behavior therapy (CBT), pharmacotherapy, and their combination therapy for acute panic disorder via network Meta-analysis.Methods:Electronic databases including PubMed, Web of Science, Embase, Cochrane Library, CNKI, and Wanfang Data were searched for relevant randomized controlled clinical trials from their inception up to March 24, 2021. The outcomes were efficacy (remission rate and response rate), acceptability and tolerability. Network Meta-analysis was adopted with random effects, and estimated relative risk (RR) for all effects of CBT, CBT plus medications, selective serotonin-reuptake inhibitors(SSRIs), serotonin-noradrenaline-reuptake inhibitors(SNRIs), tricyclic antidepressants(TCAs), monoamine oxidase inhibitors(MAOI), benzodiazepines(BZD), noradrenergic-reuptake inhibitors(NRI), and others. This study used gemtc package in R for network analysis, Review Manager for quality assessment, and GRADEPro for evidence assessment, respectively.Results:Identified citations (7 173) included 72 trials comprising 12 293 patients. The treatment measures with higher remission rate and response rate than placebo included CBT+drug( RR:2.4, 95% CI: 1.7-3.5), CBT( RR:1.8, 95% CI:1.4-2.3), SNRI( RR:1.8, 95% CI:1.5-2.2), BZD( RR:1.6, 95% CI:1.4-1.9), SSRI( RR:1.5, 95% CI:1.4-1.7), TCA( RR:1.5, 95% CI: 1.4-1.8)(remission rate in brackets); Among them, CBT combined with medications was better than SSRI, SNRI and TCA. The acceptability of BZD was better than placebo but its tolerability was inferior. Besides, the tolerability of SSRI and TCA was inferior to placebo. Conclusions:CBT combined with medications is more effective than BZD alone and the antidepressants alone. CBT combined with medications may selected the first-line treatment for panic disorder.

Objective:To reveal the mechanism of Mahuang Lianqiao Chixiaodou decoction and its disassembled formula for improving the skin barrier function in a mouse model of atopic dermatitis (AD).Methods:Sixty specific-pathogen free male BALB/c mice were randomly divided into the control group,model group,whole formula group (WF),exterior-releasing formula group (ERF),interior-clearing for-mula group (ICF),and positive control group (PC).A mouse model of AD was established using the semi-antigen 2,4-dinitrofluorobenzene induction method.The lesion scores,transepidermal water loss and pH,and skin histopathology of mice in each group were observed.The expressions of filaggrin,loricrin,and involucrin were detected by the streptavidin peroxidase immunohistochemical method and western blotting,and their mRNA expressions were detected by quantitative polymerase chain reaction.Results:Mice in the WF,ERF,ICF,and PC groups showed reduced skin lesion performance,improved histopathology,decreased skin lesion score,transepidermal water loss and pH,and upregulated ex-pressions of proteins including filaggrin,loricrin,and involucrin,and their mRNAs.The most obvious regulatory effect was observed in the WF group,followed by the ICF,ERF,and PC groups,accordingly.Conclusions:Mahuang Lianqiao Chixiaodou decoction and its disassembled formula can improve the skin barrier function in a mouse model of AD by upregulating filaggrin,loricrin,and involucrin,and their mRNA expressions,and the most optimal effect was noted in the WF group,followed by the ICF and ERF groups,which suggests that the effect of clearing heat and resolving dampness in improving the skin barrier function of AD is more obvious and is one of the key treatments for AD.

Object:This study aims to compare the efficacy, acceptability and tolerability of cognitive behavior therapy (CBT), pharmacotherapy, and their combination therapy for acute panic disorder via network Meta-analysis.Methods:Electronic databases including PubMed, Web of Science, Embase, Cochrane Library, CNKI, and Wanfang Data were searched for relevant randomized controlled clinical trials from their inception up to March 24, 2021. The outcomes were efficacy (remission rate and response rate), acceptability and tolerability. Network Meta-analysis was adopted with random effects, and estimated relative risk (RR) for all effects of CBT, CBT plus medications, selective serotonin-reuptake inhibitors(SSRIs), serotonin-noradrenaline-reuptake inhibitors(SNRIs), tricyclic antidepressants(TCAs), monoamine oxidase inhibitors(MAOI), benzodiazepines(BZD), noradrenergic-reuptake inhibitors(NRI), and others. This study used gemtc package in R for network analysis, Review Manager for quality assessment, and GRADEPro for evidence assessment, respectively.Results:Identified citations (7 173) included 72 trials comprising 12 293 patients. The treatment measures with higher remission rate and response rate than placebo included CBT+drug( RR:2.4, 95% CI: 1.7-3.5), CBT( RR:1.8, 95% CI:1.4-2.3), SNRI( RR:1.8, 95% CI:1.5-2.2), BZD( RR:1.6, 95% CI:1.4-1.9), SSRI( RR:1.5, 95% CI:1.4-1.7), TCA( RR:1.5, 95% CI: 1.4-1.8)(remission rate in brackets); Among them, CBT combined with medications was better than SSRI, SNRI and TCA. The acceptability of BZD was better than placebo but its tolerability was inferior. Besides, the tolerability of SSRI and TCA was inferior to placebo. Conclusions:CBT combined with medications is more effective than BZD alone and the antidepressants alone. CBT combined with medications may selected the first-line treatment for panic disorder.

Objective To investigate the correlation between methylation status of UNC 5C gene promoter with colorectal cancer UNC5C .Methods 54 cases of sporadic colorectal cancer and related normal mucosal tissue ,as well as 6 colon cancer cell lines and fiber cell lines(FCL) in the oncology department of the Kailuan General Hospital from February 2010 to March 2013 were selected as the research objects and performed the mRNA and methylation analysis for exploring the correlation between the netrin‐1 receptor UNC5C defects with colorectal cancer disease .Results mRNA of UNC5A and UNC5B was expressed in the detected colorectal cancer cell lines .Except FCL for UNC5C ,all of the cancer cell lines had no mRNA expression .In colorectal cancer tissue , UNC5C methylation was significantly higher than that in the normal mucosa ,the difference was statistically significant (P<0 .05) . The methylation levels of UNC5C gene in the stage Ⅰ to Ⅱ ,and the stage Ⅲ to Ⅳwere significantly higher than non‐methylation levels ,the differences were statistically significant (P<0 .05) .The correlation analysis by the Spearman method showed that the UNC5C defects was positively correlated with colorectal cancer disease (r= 0 .856 ,P< 0 .05) .Conclusion Netrin‐1 receptor UNC5C defect has certain correlation with colorectal cancer disease .

Objective To study the significance of Helicobacter pylori (Hp) infection and the expression of caudal-related homeobox transcription factor 2 (CDX2),mucin 2 (MUC2) proteins in gastric cancer,and elucidate the prognostic significance of CDX2,MUC2 proteins in gastric cancer.Methods Ninety cases of gastric cancer were available for the study.Hp was studied by Warthin-Starry bacterium stain and CDX2 and MUC2 proteins were studied by immunohistochemical stain.Results There was no Hp infection and no expression of CDX2 and MUC2 proteins in normal gastric mucosa.The positive rate in the cases of intestinal-type gastric cancer was 67.86%(38/56) for Hp,75.00% (42/56) for CDX2,and 66.07%(37/56) for MUC2,respectively,and it was higher than that in the cases of diffuse-type gastric cancer,which was 35.29%(12/34),41.18%(14/34) and 38.24%(13/34) respectively (P<0.01 or < 0.05).In intestinaltype gastric cancer,the positive rate of CDX2 and MUC2 proteins was also higher in Hp positive cases than that in Hp negative cases,and it was the same as CDX2 protein in diffuse-type gastric cancer.Higher positive rates of CDX2 and MUC2 proteins were observed in the cases of early gastric cancer than those in the cases of advanced gastric cancer (P < 0.01 or < 0.05),also it was observed in the cases of no lymph node metastasis than in the cases of lymph node metastasis (P < 0.01 or < 0.05).Moreover,the positive rate of CDX2 protein in the cases of stage Ⅰ～Ⅱwas higher than that in the cases of stage Ⅲ～Ⅳ (P < 0.05).Conclusions Hp infection and the expression of CDX2 and MUC2 proteins are related with gastric cancer,and has close relationship with intestinal-type gastric cancer.The expressions of CDX2 and MUC2 proteins may inhibit the infiltration and metastasis of gastric cancer.

AIM To study whether cyproheptadine(Cyp) affects endocrine functions in reproductive system with gender difference. METHODS Sixty SD rats were randomly distributed into 3 groups according to gender, respectively, and they were administered NS(5 mL*kg-1*d-1), Cyp 2.4, 4.8 mg*kg-1*d-1 accordingly by ig for 14 d or 21 d. The serum levels of luteinizing hormone(LH), follicle stimulating hormone(FSH), testosterone(T), progesterone(P), estrodiol(E2) were measured by radio-immunoassay and the ultrastructure of gonadotropin-releasing hormone(GnRH) cells, gonadotropin cells, Leydig cells, Sertoli cells, luteal cells, granulocytes and so on were observed by electronmicroscopy and microscopy. The calmodulin(CaM) mRNA expression in hypothalamus-pituitary-testis axis(HPTA) was detected by reverse transcriptase-polymerase chain reaction. RESULTS Cyp(2.4 mg*kg-1*d-1×14 d, ig) increased serum LH concentration while decreased serum FSH, P concentrations in female rats. Cyp(4.8 mg*kg-1*d-1×14 d, ig)increased serum LH, T concentrations in males, and increased serum LH concentration while decreased serum FSH, E2 and P concentrations significantly in females. The retrograde changes of ultrastructure were observed in part of gonadotropin and ovary endocrine cells, while a stimulating one in testis endocrine cells. CaM mRNA expression levels were elevated in testis but not in hypothalamus and pituitary in male rats. CONCLUSIONCyp had a negative effect on endocrine function in females, but a positive one in males. The ultrastructure showed relevant changes in target gland. Cyp promoted CaM mRNA expression in testis,which had close connections with Cyp′s stimulative effect in HPTA.

AIM To study the effects of cyproheptadine (Cyp) on endocrine functions and pituitary adrenalcortex axid in rats and its mechanism METHODS Radioimmunoassay(RIA)was used to measure the serum levels of ACTH and cortisone Ultramicrostructure of the ACTH cells and adrenalcortex and the calmodulin(CaM) mRNA expression of the pituitary and adrenalcortex were observed by electron microscope and the quantity PCR technique RESULTS Cyp 2 3 and 4 6 mg?kg -1 ?d -1 , ig for 14 d had significantly reduced the serum levels of ACTH and cortisone ( P