Chemoprotective effect of arbutin on azoxymethane-induced aberrant crypt foci in rat colon via modulation of PCNA/Bax protein

Arbutin is utilized in traditional remedies to cure numerous syndromes because of its anti-microbial, antioxidant, and anti-inflammatory properties. This study aimed to evaluate chemopreventive effects of arbutin on azoxymethane (AOM)-induced colon aberrant crypt foci (ACF) in rats. Five groups of rats were used: normal control group (rats injected hypodermically with sterile phosphate-buffered saline once per week for two weeks) and groups 2-5, which were subcutaneously inoculated with 15 mg/kg AOM once a week for two weeks. AOM control and 5-fluorouracil (5-FU) control groups were fed 10% Tween orally daily for 8 weeks using a feeding tube. The treated groups were fed 30 and 60 mg/kg arbutin every day for 2 months. ACF from the AOM control group had aberrant nuclei in addition to multilayered cells and an absence of goblet cells. The negative control group displayed spherical cells and nuclei in basal positions. Histological examination revealed a reduced number of AFC cells from colon tissues of the 5-FU reference group. Arbutin-fed animals showed down-regulation of proliferating cell nuclear antigen (PCNA) and up-regulation of Bax protein compared to AOM control. Rats fed with arbutin displayed a significant increase of superoxide dismutase (SOD) and catalase (CAT) activities in colon tissue homogenates compared to the AOM control group. In conclusion, arbutin showed therapeutic effects against colorectal cancer, explained by its ability to significantly decrease ACF, down-regulate PCNA protein, and up-regulate Bax protein. In addition, arbutin significantly increased SOD and CAT, and decreased malondialdehyde (MDA) levels, which might be due to its anti-proliferative and antioxidant properties.

Relationship between expressions of P21,P27 and PCNA in glomerular mesangial tissue and poor renal prognosis in patients with IgA nephropathy / 中国免疫学杂志

Objective:To explore the relationship between the expressions of P21,P27 and proliferating cell nuclear antigen(PCNA)in glomerular mesangial tissue and poor renal prognosis in patients with immunoglobulin A(IgA)nephropathy.Methods:A total of 145 patients with IgA nephropathy treated in Xiaogan Central Hospital from April 2017 to August 2019 were selected as the research object.The expressions of P21,P27 and PCNA in glomerular mesangial tissue were detected by immunohistochemistry.All patients were followed up for 24 months,and the prognosis were counted.The expressions of P21,P27 and PCNA in glomerular mesangial tissue of patients with different prognosis were compared and the influencing factors of poor prognosis in patients with IgA nephropathy were analyzed by Logistic regression analysis.Results:The expression rates of P21,P27 and PCNA positive cells in glomerular mesangial tissue of patients with IgA nephropathy were(38.69±6.83)%,(55.94±8.08)%,(33.47±5.72)%,respectively.The incidence rete of poor prognosis in patients with IgA nephropathy was 17.24%,and the expression rates of P21 and PCNA positive cells in glomerular mesangial tissue of patients with poor prognosis were higher than those in good prognosis group(P<0.05),while the expression rate of P27 positive cells was lower than that in good prognosis group(P<0.05).Logistic multiple regression analysis showed that elevated diastolic blood pressure,increased 24 h proteinuria,mesangial cell proliferation,segmental glomerulosclerosis,renal tubular atrophy/interstitial fibrosis,crescentic body,increased expression rates of P21 and PCNA positive cells and decreased expression rate of P27 positive cells were all risk factors affecting the poor prognosis of patients with IgA nephropathy(P<0.05).Conclusion:There are positive expressions of P21,P27 and PCNA in glomerular mesangial tissue of IgA nephropathy.The expression rates of P21 and PCNA positive cells in glomerular mesangial tissue of of patients with poor prognosis of IgA nephropathy are higher than those with good prognosis,while the expression rate of P27 protein positive cells is lower than those with good prognosis,which are risk factors for poor prognosis of patients with IgA nephropathy.

Gentiopicroside injection promotes the healing of pressure injury wounds by upregulating the expression of bFGFR1 / A injeção de gentiopicrosídeo promove a cicatrização de feridas de lesão por pressão pela suprarregulação da expressão de bFGFR1 / La inyección de gentiopicrósido promueve la cicatrización de heridas de lesión por presión al aumentar la expresión de bFGFR1

ABSTRACT Objective: To observe the therapeutic effect of gentiopicroside, as the main component of Gentianaceae, on wounds in pressure injury (PI) model rats and explore its mechanism. Method: Male Sprague Dawley rats were randomly divided into control group, model group and gentiopicroside groups (50, 100 and 200 mg·kg-1·d-1 for 9 consecutive days). The mice's skeletal muscle fibroblast line NOR-10 cells were collected after being treated with gentiopicroside (0.2~5.0 M) and basic fibroblast growth factor receptor 1 (bFGFR1) inhibitor (5.0 M SU5402) for 7 days. Results: Compared to the model group, the gentiopicroside groups showed significantly increased wound healing rates, reduced inflammatory cells in the wound tissues, and significantly increased expression levels of proliferating cell nuclear antigen (PCNA) and bFGFR1, accompanied by increased proliferation of new myofibroblasts. Gentiopicroside upregulated the mRNA expression of bFGFR1 and PCNA in NOR-10 cells in a dose-dependent manner; however, SU5402 reversed the effect of gentiopicroside. Conclusion: Gentiopicroside may promote myofibroblast proliferation by upregulating the expression of bFGFR1 and PCNA and ultimately accelerating the healing of PI wounds.

RESUMO Objetivo: Observar o efeito terapêutico do gentiopicrosídeo como principal componente das Gentianáceas em feridas de lesão por pressão (LP) em modelos de ratos e explorar seu mecanismo. Métodos: Ratos Sprague Dawley machos foram divididos aleatoriamente em grupo controle, grupo modelo e grupos gentiopicrosídeo (50, 100 e 200 mg·kg-1·d-1 por 9 dias consecutivos). As células NOR-10 da linha de fibroblastos do músculo esquelético de camundongos foram coletadas após serem tratadas com gentiopicrosídeo (0,2~5,0 μM) e inibidor do receptor 1 do fator de crescimento fibroblástico básico (bFGFR1) (5.0 μM SU5402) por 7 dias. Resultados: Em comparação com o grupo modelo, os grupos gentiopicrosídeo apresentaram taxas de cicatrização de feridas significativamente maiores, menos células inflamatórias nos tecidos da ferida e níveis de expressão de antígeno nuclear de proliferação celular (PCNA) e bFGFR1 significativamente maiores, acompanhados por aumento da proliferação de novos miofibroblastos. O gentiopicrosídeo regulou positivamente a expressão de mRNA de bFGFR1 e PCNA em células NOR-10 de maneira dependente da dose, enquanto o SU5402 reverteu o efeito do gentiopicrosídeo. Conclusão: O gentiopicrosídeo pode promover a proliferação de miofibroblastos, suprarregulando a expressão de bFGFR1 e PCNA e, em última análise, acelerando a cicatrização de feridas de LP.

RESUMEN Objetivo: Observar el efecto terapéutico del gentiopicrósido como componente principal de la Gentianaceae en heridas por lesión por presión (LP) en modelos de ratas y explorar su mecanismo. Método: Se dividieron aleatoriamente ratas macho Sprague Dawley en grupo control, grupo modelo y grupos gentiopicrósido (50, 100 y 200 mg·kg-1·d-1 durante 9 días consecutivos). Se recogieron células NOR-10 de la línea de fibroblastos de músculo esquelético de ratón después de ser tratadas con gentiopicrósido (0.2~5.0 μM) y un inhibidor del receptor 1 del factor de crecimiento de fibroblastos básico (bFGFR1) (5.0 μM SU5402) durante 7 días. Resultados: En comparación con el grupo modelo, los grupos de gentiopicrósido mostraron tasas de curación de heridas significativamente más altas, menos células inflamatorias en los tejidos de la herida y niveles de expresión significativamente mayores del antígeno nuclear de proliferación celular (PCNA) y bFGFR1, acompañados de una mayor proliferación de nuevos miofibroblastos. El gentiopicrósido podría regular positivamente la expresión de ARNm de bFGFR1 y PCNA en células NOR-10 de manera dependiente de la dosis, sin embargo, SU5402 revirtió el efecto del gentiopicrósido. Conclusión: El gentiopicrósido puede promover la proliferación de miofibroblastos al aumentar la expresión de bFGFR1 y PCNA y, en última instancia, acelerar la cicatrización de las heridas de LP.

Mechanism of CD137 signal regulating P53/P21 pathway to promote senescence of vascular smooth muscle cells / 中华老年心脑血管病杂志

Objective To explore the mechanism by which CD137 signal regulates the aging of vas-cular smooth muscle cells(VSMCs).Methods Thirty 8-week-old male C57BL/6J mice were ran-domly divided into a young group(8 weeks old)and an aged group(80 weeks old),with 30 mice in each group.After corresponding periods of feeding,the mice were euthanized,and the plasma and aortic blood vessels were isolated.In the cell experiments,normal VSMCs were divided into a control group,bleomycin(BLM)group,combined agonist group,and combined inhibitor group.The cellular senescence level of VSMCs was assessed using a cellular senescence β-galactosidase staining kit.Western blotting and PCR were employed to examine the expression of senescence-related proteins in tissues and cells,while ELISA was utilized to measure the expression of senes-cence-related inflammatory factors.Results The expression of CD137 and γ-H2AX in the aorta was significantly higher,while that of PCNA was obviously lower in the aged group than the young group(P＜0.05).The plasma level of CD137 was notably higher in the aged group than the young group(154.0±4.1 pg/ml vs 98.0±2.3 pg/ml,P＜0.05).Compared with the normal control group,there were significantly more aged VSMCs in the BLM group(P＜0.05).While,treatment of combined agonist resulted in larger amount of aged VSMCs when compared with the BLM group(P＜0.05),which was reversed by combined inhibitor treatment(P＜0.05).The levels of TNF-α,IL-6 and IL-1β were significantly elevated in the BLM group than the normal control group(P＜0.05).The combined agonist group had even higher levels of TNF-α,IL-6,and IL-1βthan the BLM group(P＜0.05),but the levels were decreased in the combined inhibitor group(P＜0.05).Compared with the normal control group,the expression of Bcl-2,γ-H2AX,P53,and P21 were significantly increased in the BLM group,combined agonist group,and combined inhibi-tor group,while that of PCNA was significantly decreased(P＜0.05).Compared with the BLM group,the expression of P53 and P21 in the combined agonist group showed an increase(P＜0.05),and the expression of P53 was significantly decreased in the combined inhibitor group(P＜0.05).Conclusion CD137 signal regulates the P53/P21 pathway to promote VSMC aging.

The relationship between the expression levels of monocyte chemotactic protein-1, proliferating cell nuclear antigen, nuclear factor-κBp65 and pregnancy outcome in pregnant women with gestational diabetes mellitus / 中国医师进修杂志

Objective:To investigate the placenta tissue expression levels of monocyte chemotactic protein-1 (MCP-1), nuclear factor-κBp65 (NF-κBp65) and proliferating cell nuclear antigen (PCNA) in pregnant women with gestational diabetes mellitus (GDM), and to analyze their correlation with pregnancy outcomes.Methods:The clinical data of 124 pregnant women with GDM from May 2020 to December 2021 in PLA Northern Theater General Hospital were retrospectively analyzed. Among them, 62 pregnant women were willing to receive treatment (treatment group), while 62 pregnant women were unwilling to receive treatment (untreated group). In addition, 80 healthy pregnant women in the same period were selected as the healthy control group. The natural birth rate, neonatal Apgar score and the incidences of macrosomia, neonatal hypoglycemia, neonatal hyperbilirubinemia were record. The placenta tissue expression levels of MCP-1, NF-κBp65 and PCNA were detected by immunohistochemical.Results:The natural birth rate in untreated group was significantly lower than that in treatment group and healthy control group: 24.19% (15/62) vs. 75.81% (47/62) and 88.75% (71/80), the natural birth rate in treatment group was significantly lower than that in healthy control group, and there was statistical difference ( P<0.05). The Apgar score in untreated group was significantly lower than that in treatment group and healthy control group: (8.45 ± 2.02) scores vs. (9.46 ± 2.59) and (9.71 ± 3.21) scores, the incidences of macrosomia, neonatal hypoglycemia and neonatal hyperbilirubinemia were significantly higher than those in treatment group and healthy control group: 35.48% (22/62) vs. 11.29% (7/62) and 3.75% (3/80), 29.03% (18/62) vs. 8.06% (5/62) and 2.50% (2/80), 24.19% (15/62) vs. 9.68% (6/62) and 2.50% (2/80), and there were statistical differences ( P<0.05); there were no statistical difference in the indexes treatment group and healthy control group ( P>0.05). The positive expression rates of MCP-1, NF-κBp65 and PCNA in untreated group were significantly higher than those in treatment group and healthy control group: 72.58% (45/62) vs. 25.81% (16/62) and 12.50% (10/80), 69.35% (43/62) vs. 27.43% (17/62) and 13.75% (11/80), 69.35% (43/62) vs. 24.19% (15/62) and 11.25% (9/80), the indexes in treatment group were significantly higher than those in healthy control group, and there were statistical differences ( P<0.05). Conclusions:The placenta tissue expression levels of MCP-1, NF-κBp65 and PCNA in pregnant women with GDM are associated with adverse pregnancy outcomes. After active treatment, the positive rates of the three indexes in pregnant women with GDM significantly decrease, and the prognosis got improved.

Protective effect of nebivolol on rat ovary exposed to deltamethrin toxicity

Purpose: We aimed to investigate the antioxidant activity of nebivolol against possible damage to the ovarian tissue due to the application of deltamethrin as a toxic agent, by evaluating histopathological proliferating cell nuclear antigen (PCNA) and tumor necrosis factor-alpha (TNF-α) signal molecules immunohistochemically. Methods: The animals were divided into three groups as control, deltamethrin and deltamethrin + nebivolol groups. Vaginal smears were taken after the animals were mated and detected on the first day of pregnancy. After the sixth day, deltamethrin (0.5 mL of 30 mg/kg BW undiluted ULV), and 2 mL of sterile nebivolol solution were administered intraperitoneally every day for 6-21 periods. After routine histopathological follow-up, the ovarian tissue was stained with hematoxylin and eosin stain. Results: Control group showed normal histology of ovarium. In deltamethrin group, hyperplasic cells, degenerative follicles, pyknotic nuclei, inflammation and hemorrhagic areas were observed. Nebivolol treatment restored these pathologies. Deltamethrin treatment increased TNF-α and PCNA reaction. However, nebivolol decreased the expression. Conclusions: It was thought that deltamethrin toxicity adversely affected follicle development by inducing degeneration and apoptotic process in preantral and antra follicle cells, and nebivolol administration might reduce inflammation and slow down the apoptotic signal in the nuclear phase and regulate reorganization.

Effect of Inhibiting Upstream Transcription Factor 2 Expression on Proliferation and Apoptosis of Gastric Cancer BGC-823 Cells / 肿瘤防治研究

Objective To investigate the effect of upstream transcription factor 2 (USF2) on the proliferation and apoptosis of human gastric cancer BGC-823 cells. Methods Lipofectamine?3000 transfection reagent was used to transfect USF2 siRNA into BGC-823 cells (siRNA-USF2 group). Blank control and negative control (siRNA-NC) groups were also prepared. The mRNA and protein expression levels of USF2 in transfected BGC-823 cells were detected by real-time fluorescence quantitative PCR and Western blot, respectively. The proliferation and clone formation abilities of BGC-823 cells in each group were investigated by CCK-8 and plate cloning assay. The apoptosis of gastric cancer cells was examined by flow cytometry. The expression levels of PCNA and apoptosis-related proteins Bax and Bcl-2 in BGC-823 cells were measured by Western blot. Results Compared with those in the blank control and siRNA-NC groups, the mRNA and protein expression levels of USF2 significantly decreased in the siRNA-USF2 group (P < 0.05). At 72 h after transfection, the absorbance in the siRNA-USF2 group was lower than that in the blank control group (P < 0.05). Compared with that in the blank control and siRNA-NC groups, the number of BGC-823 cell clones significantly decreased in the siRNA-USF2 group (P < 0.05). The apoptosis rate of BGC-823 cells significantly differed among the blank control, siRNA-NC, and siRNA-USF2 groups (P < 0.05). Compared with those in the blank control and siRNA-NC groups, the expression of PCNA and Bcl-2 protein decreased and that of Bax protein increased in the siRNA-USF2 group (P < 0.05). Conclusion Inhibiting USF2 expression can suppress the proliferation of human gastric cancer cells and induce their apoptosis. USF2 inhibitors may have important value in the treatment of gastric cancer.

Effect of silencing HDAC9 on the proliferation and differentiation of periodontal ligament stem cells / 口腔疾病防治

Objective @#To investigate the effect of silencing histone deacetylase 9 (HDAC9) expression on the proliferation and osteogenic differentiation of periodontal ligament stem cells (PDLSCs).@*Methods@# PDLSCs were isolated, cultured and identified in vitro. An siRNA construct specific for HDAC9 was transfected into PDLSCs (siHDAC9 group), and a nontargeting siRNA was used as a control (siNC group). The interference effect was determined by qRT-PCR. The cell cycle progression of PDLSCs was detected using flow cytometry. The proliferation activity of PDLSCs was detected via CCK-8 assay. Western blotting was used to detect the protein expression of proliferating cell nuclear antigen (PCNA). The mRNA expression of runt-related transcription factor 2 (RUNX2) and alkaline phosphatase (ALP) was investigated by qRT-PCR. The protein expression of RUNX2 was detected by western blotting. In addition, the formation of mineralized nodules was assessed by alizarin red staining. @*Results@#Compared with that in the siNC group, the mRNA expression of HDAC9 in the siHDAC9 group was lower (P < 0.01). Moreover, compared with those in the siNC group, the proliferation index (P<0.01), proliferation activity (P<0.05) and protein expression of PCNA (P<0.01) in the siHDAC9 group were all increased. Compared with the siNC group, the siHDAC9 group exhibited higher mRNA expression of RUNX2 and ALP (P < 0.05), and the protein expression of RUNX2 showed the same results (P < 0.01). The results of alizarin red staining showed that compared to the siNC group, the siHDAC9 group formed more mineralized nodules.@* Conclusion@#Silencing HDAC9 expression can promote the proliferation and osteogenic differentiation of PDLSCs.

Expression of minichromosome maintenance protein 6 (MCM6) in colon cancer tissues and its correlation with proliferating cell nuclear antigen (PCNA) / 国际生物医学工程杂志

Objective:To analyze the expression level of minichromosome maintenance protein 6 (MCM6) in colon cancer tissues, the correlation between the expression level of MCM6 and the clinicopathological characteristics of colon cancer patients, and the correlation between MCM6 and PCNA expression.Methods:The expression levels of MCM6 in different tumor tissues were analyzed based on the Human Protein Atlas (HPA) database. The expression levels and correlations of MCM6 and PCNA in colon cancer tissues were analyzed based on The Cancer Genome Atlas (TCGA) database and immunohistochemical experiments. The correlation between MCM6 expression level and clinical characteristics of colon cancer patients was analyzed. The correlation between MCM6 and PCNA expression in colon cancer was analyzed based on TCGA database and Gene Expression Profile Interaction Analysis (GEPIA) database.Results:Bioinformatics analysis and immunohistochemical results confirmed that MCM6 was highly expressed in colon cancer tissues, and its expression level was correlated with the tumor stage of patients ( P=0.01). In colon cancer, the expression of MCM6 and PCNA was correlated with statistical significance ( P<0.05). Conclusions:MCM6 is highly expressed in colon cancer tissue and is related to the clinical characteristics of patients, suggesting that MCM6 can be used as a potential marker of colon cancer.

Effect and Mechanism of "Liver-Soothing and Spleen-Invigorating" Therapies Against Liver Injury in Rats： An Exploration Based on TGR5 and Intestinal Mucosal Barrier Function / 中国实验方剂学杂志

ObjectiveTo explore the effects of different treatment methods of "soothing liver， invigorating spleen， soothing liver and invigorating spleen， soothing liver first and then soothing liver and invigorating spleen， as well as invigorating spleen first and then soothing liver and invigorating spleen" on liver depression combined with liver injury in rats and their action mechanisms. MethodA six-week rat model of liver depression combined with liver injury was established by restraint stress and subcutaneous injection of carbon tetrachloride （CCl4， 5.89 g·kg-1， once every three days）. At the same time， the drugs were given by gavage. Forty-eight male SD rats of clean grade were randomly divided into eight groups， namely the normal group， model group， bicyclol （0.2 g·kg-1） group， Sinisan （4.32 g·kg-1） group， Liu Junzitang （9.26 g·kg-1） group， Chaishao Liu Junzitang A （Chai A， soothing liver and invigorating spleen，13.57 g·kg-1） group， Chaishao Liu Junzitang B （Chai B， soothing liver first and then soothing liver and invigorating spleen， 13.57 g·kg-1） group， and Chaishao Liu Junzitang C （Chai C， invigorating spleen first and then soothing liver and invigorating spleen， 13.57 g·kg-1） group， with six rats in each group. The pathological changes in liver and colon tissues of each group were observed under light microscope and electron microscope. The serum biochemical indexes of the liver were detected using an automatic biochemical analyzer. The relative mRNA expression levels of Takeda G protein-coupled receptor 5 （TGR5） and intestinal mucosal zona occluden-1 （ZO-1）， Occludin， and Claudin-1 in the liver and colon were detected by reverse-transcription polymerase chain reaction （RT-PCR）. The positive expression rate of proliferating cell nuclear antigen （PCNA） in the colon was detected by immunohistochemistry. ResultCompared with normal group， the model group exhibited significantly elevated serum alkaline phosphatase （ALP）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， and direct bilirubin （DBIL） （P<0.01）， lowered TGR5 mRNA expression in liver tissue， up-regulated TGR5 mRNA expression in the colon tissue （P<0.05，P<0.01）， and down-regulated ZO-1， Occludin， and tight junction protein-1 (Claudin-1) mRNA expression and PCNA in the colon tissue （P<0.01）. Compared with the model group， bicyclol and Chai C remarkably decreased the levels of serum ALP， ALT， AST， TBIL， and DBIL （P<0.05，P<0.01）， while Liu Junzitang， Chai A， Chai B， and Chai C significantly up-regulated the TGR5 mRNA expression in the liver and down-regulated its expression in the colon （P<0.01）. Bicyclol， Chai A， Chai B， and Chai C enhanced the ZO-1 and Claudin-1 mRNA expression in the colon （P<0.05，P<0.01）. Bicyclol， Sinisan， and Chai C increased PCNA expression （P<0.01）. The comparison with the Chai C group showed that the TGR5 mRNA expression in the liver and ZO-1 mRNA expression in the colon of the bicyclol and Sinisan groups were lower， whereas the TGR5 mRNA expression in the colon was higher （P<0.01）. However， the PCNA expression in the colon of the Liu Junzitang and Chai B groups declined significantly （P<0.05）. ConclusionIn the presence of liver injury， invigorating spleen first helps to relieve the liver injury， and the efficacy of "spleen-invigorating" therapy in increasing the intestinal mucosal tight junction proteins and improving the gastrointestinal function is related to its activation of TGR5 to improve the intestinal mucosal barrier function， promote the renewal of intestinal stem cells， and drive the regeneration after injury.

Cohort study of changes in expression of three serum cytokines in patients with advanced non-small cell lung cancer after intensity-modulated radiotherapy combined with chemotherapy / 中华放射肿瘤学杂志

Objective:To investigate the changes of the expression levels of serum proliferating cell nuclear antigen (PCNA), tumor-specific growth factor (TSGF), soluble E-cadherin (SE-CAD) and the relationship with clinical prognosis of advanced non-small cell lung cancer (NSCLC) patients treated with intensity-modulated radiotherapy combined with chemotherapy.Methods:Eighty-four patients (29 cases of Ⅲ A, 30 Ⅲ B and 25 Ⅳ) with advanced NSCLC treated in our hospital from January 2016 to January 2018 were selected, and all patients were given with intensity-modulated radiotherapy combined with chemotherapy. The expression levels of serum PCNA, TSGF, and SE-CAD were compared among different TNM stages and before and after treatment. The serum PCNA, TSGF, SE-CAD levels were compared among patients with different clinical efficacy. The relationship between serum PCNA, TSGF and SE-CAD levels and clinical efficacy was assessed by Logistic regression analysis. The survival analysis was performed with Kaplan- Meier method. Results:The expression levels of serum PCNA, TSGF and SE-CAD before treatment in stage Ⅳ patients were significantly higher than those in stage Ⅲ B and Ⅲ A patients (584.11±60.25 pg/ml vs. 531.06±51.37 pg/ml and 477.54±46.49 pg/ml, 96.13±7.54 U/ml vs. 8.52±5.91 U/ml and 82.41±5.0 U/ml, 3.02±0.26 ng/ml vs. 2.87±0.22 ng/ml and 2.71±0.15 ng/ml, all P<0.05), and the serum levels of three cytokines in Ⅲ B stage patients were significantly higher than those in their Ⅲ A stage counterparts (all P<0.05). After treatment, the serum levels of PCNA, TSGF and SE-CAD were significantly lower than those before treatment (396.11±50.23 pg/ml vs. 528.37±75.09 pg/ml, 74.81±4.72 U/ml vs. 88.68±6.13 U/ml, 1.92±0.24 ng/ml vs.2.86±0.31 ng/ml, all P<0.05). At 18 months after treatment, the serum levels of PCNA, TSGF and SE-CAD in surviving patients were significantly lower than those of dead patients (332.51±54.32 pg/ml vs. 444.92±60.07 pg/ml, 70.59±6.20 U/ml vs. 78.05±8.44 U/ml, 1.71±0.24 ng/ml vs. 2.08±0.27 ng/ml, all P<0.05). The serum levels of PCNA, TSGF and SE-CAD were significantly associated with clinical prognosis (all P<0.05). Among 84 NSCLC patients, the objective response rate after treatment was 29%(24/84). The survival curves in patients with high expression levels of serum PCNA, TSGF and SE-CAD were significantly lower than those in the low-expression group (all P<0.05). Conclusion:Serum PCNA, TSGF and SE-CAD are highly expressed in patients with advanced NSCLC, which are closely correlated with clinical staging and prognosis and contribute to predicting survival status.

Ultrasonographic features of cervical lymph node metastasis and their relationship with the expression of cell proliferation-associated nuclear antigen / 中国基层医药

Objective:To observe the ultrasonographic features of cervical lymph node metastasis and analyze their relationship with the expression of cell proliferation-associated nuclear antigen (Ki-67).Methods:The clinical data of 92 patients with cervical lymph node metastasis who received treatment in Zhejiang Quhua Hospital, China between June 2017 and July 2019 were retrospectively analyzed. The correlation between ultrasonographic features of cervical lymph node metastasis and Ki-67 expression was analyzed.Results:Among the 92 patients, 158 metastatic lymph nodes were confirmed by pathological examination. The main ultrasonic imaging features were the length ratio of long diameter to short diameter < 2 in 119 (75.32%) lymph nodes, disappearance of portal hyperechoic signal in 127 (80.38%) lymph nodes, punctate hyperechoic signal in 108 (68.35%) lymph nodes, cystic degeneration in 57 (36.08%) lymph nodes, mixed types of peripheral blood flow signal in 124 (78.48%) lymph nodes, microcalcification in 123 (77.85%) lymph nodes. The length ratio of long diameter to short diameter < 2, punctate hyperechoic signal and mixed types of peripheral blood flow signal were correlated with high expression of Ki-67 ( χ2 = 24.252, 15.732, 17.033, all P < 0.05). Conclusion:High expression of Ki-67 is correlated with the length ratio of long diameter to short diameter < 2, punctate hyperechoic signal and mixed types of peripheral blood flow signal in cervical lymph node metastasis.

Expressions of cytokeratin 14 and proliferating cell nuclear antigen in the Hertwig's epithelial root sheath of a Vps4b knockout mouse / 华西口腔医学杂志

OBJECTIVES@#The effect of Vps4b gene mutation on the expressions of cytokeratin 14 (CK14) and proliferating cell nuclear antigen (PCNA) in the Hertwig's epithelial root sheath (HERS) is investigated.@*METHODS@#The bilateral mandibular tissues of mouse on postnatal days 5, 9, 11, 15, and 19 were removed. The mandibular first molar tissue sections were obtained after paraffin embedding. The CK14 and PCNA expressions in the epithelial root sheath of the normal mouse and Vps4b knockout mouse were compared through immunohistochemistry.@*RESULTS@#On postnatal day 5, the normal mouse began to form HERS and had a strong positive PCNA expression in the HERS cells; on postnatal day 9, the HERS structure was continuous, and PCNA was positive in the HERS cells; on postnatal day 11, a small portion of HERS began to break, and PCNA was weakly positive in the HERS cells; on postnatal day 15, HERS continued to fracture; PCNA was weakly and positively expressed in the HERS cells on the root surface; on postnatal day 19, the tooth root reached normal physiological length, and PCNA was positively expressed in the HERS cells of the terminal part. Similar to the normal mouse, the gene knockout mouse also formed a HERS structure on postnatal day 5. However, HERS began to break on postnatal day 9. On postnatal day 19, only a few fragments of HERS were found on the root surface, and the root development was immature. Moreover, the expression intensity of PCNA in the gene knockout mouse was decreased.@*CONCLUSIONS@#The Vps4b gene mutation may change the CK14 and PCNA expressions, leading to abnormal root development.

Expression of MCM4/PCNA in gastric cancer and its potential as a potential biomarker / 国际生物医学工程杂志

Objective:To analyze the expressions of minichromosome maintenance protein 4(MCM4) and proliferating cell nuclear antigen (PCNA) in gastric cancer tissues, to explore the relationship between MCM4/PCNA and gastric cancer, and to investigate the possibility of MCM4/PCNA as a potential biomarker for gastric cancer.Methods:Bioinformatics methods were used to analyze the mRNA expressions of MCM4 and PCNA in gastric cancer tissues and adjacent normal tissues. The clinicopathological data of 69 patients with gastric cancer who underwent surgery were retrospectively analyzed. The expression levels of MCM4 and PCNA in gastric cancer tissues and adjacent normal tissues were detected by immunohistochemistry, and their relationship with the clinicopathological characteristics of gastric cancer patients was analyzed.Results:The mRNA levels of MCM4 and PCNA in gastric cancer tissues are significantly higher than those in adjacent normal tissues (all P<0.05). The expression of MCM4 is correlated with the tumor size of gastric cancer ( P=0.037), but there is no significant correlation with gender, age and tumor grade (all P>0.05). Both MCM4 and PCNA proteins are highly expressed in gastric cancer patients. Conclusions:The expression levels of MCM4 and PCNA have a clear correlation with the occurrence of gastric cancer. MCM4 and PCNA are expected to be potential biomarkers for the diagnosis and treatment of gastric cancer.

PCNA Confers the Conversion of Hepatitis B Virus Relaxed Circular DNA to Covalently Closed Circular DNA by Its DNA Binding Domain / 中国生物化学与分子生物学报

Chronic hepatitis B virus (HBV) infection is a primary cause for liver cancer. And the main challenge of curing hepatitis B is the elimination of the stable covalently closed circular DNA (cccDNA) of the viral genome. The formation of HBV cccDNA requires the filling of single-stranded region and the ligation of nicks in relaxed circular DNA (rcDNA) strands. Previously, our group reported that proliferating cell nuclear antigen (PCNA) was involved in the formation of HBV cccDNA. However, the underlying mechanism of the conversion of HBV rcDNA to cccDNA is poorly understood. In the present study, we aim to explore the mechanism by which PCNA contributes to the conversion of HBV rcDNA to cccDNA. Our data showed that PCNA was involved in the process of HBV rcDNA repair. The knockout of PCNA by the CRISPR/Cas9 system remarkably blocked the conversion of HBV rcDNA to cccDNA, while the ectopic expression of PCNA could effectively rescue the event (P<0. 001). Knockout of PCNA significantly slowed down the conversion kinetics of HBV rcDNA to cccDNA (P<0. 01). Mechanically, the DNA binding domain of PCNA was required for the process of HBV rcDNA repair to cccDNA (P<0. 01). Thus, we conclude that PCNA confers the conversion of HBV rcDNA to cccDNA by its DNA binding domain. Clinically, PCNA might serve as a novel target for antiviral therapy.

Nesiritide protects endothelial function after balloon-induced trauma in the iliac artery in rabbits

To assess the effect of nesiritide on the endothelial function of iliac arteries following endothelia trauma. Right iliac artery trauma was created with a balloon catheter. Ten rabbits were treated with a 4-week subcutaneous injection of nesiritide at a fixed daily dose of 0.1mg/kg. Ten rabbits received daily normal saline injection. Plasma endothelin 1 (ET-1), nitric oxide (NO), and Von Willebrand Factor (vWF) were measured before and after the therapies. Tissue proliferating cell nuclear antigen (PCNA) was measured after the treatment. After the treatment, in the therapeutic group, the area under internal elastic membrane and the residual lumen area were higher than in the normal saline group (P <0.05). The plasma levels of ET-1 (91.6±6.8 vs 114.9±6.3 ng/L, P =0.001), vWF (134.6±10.8% vs 188.8±10.4%, P =0.001) and the ratio of PCNA positive expression (11.7±4.2% vs 36.2±11.4%, P =0.005) in the therapeutic group was lower than in the normal saline group, while the plasma levels of NO was higher (89.7±9.3 vs 43.5±5.3 µmol/L, P =0.001). Nesiritide inhibited remodeling of rabbit iliac artery following endothelial trauma. The inhibition of vascular remodeling may be related to the alleviated endothelial dysfunction and reduced expression of tissue proliferating cell nuclear antigen

Anti-scarring effect of rapamycin following filtering surgery in rabbit eyes / 南方医科大学学报

OBJECTIVE@#To study the effect of rapamycin on scar formation in rabbit eyes following filtering operation and explore the possible mechanism.@*METHODS@#Ninety-six healthy adult rabbits were subjected to trabeculectomy of the left eye and subsequently randomly divided into 4 groups (=24) for treatment with castor oil (control) or rapamycin (1%, 3%, or 5%) eye drops of the operated eyes 4 times a day. The morphology and function of the filtering blebs of the rabbits were compared at 7, 14, 21 and 28 days after the operation; at each of the time points, 6 rabbits from each group were euthanized for detection of expressions of proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) in the tissues in the surgical area using immunohistochemistry. Cultured rabbit subconjunctival fibroblasts (RTFSs) were treated with different concentrations of rapamycin (0.06, 0.25, 1, and 4 mg/L) and the cell apoptosis was detected using flow cytometry.@*RESULTS@#In the first, second and third weeks after the operation, the rate of functional follicle formation was significantly higher in the 3 rapamycin groups than in the control group ( < 0.05), and the number of α- SMA-positive fibroblasts decreased over time in the 3 rapamycin groups. In cultured RTFSs, treatment with rapamycin at different concentrations resulted in increased apoptosis of the cells, and rapamycin above 0.25 mg/L significantly increased the cell apoptosis in a dose-dependent manner.@*CONCLUSIONS@#Rapamycin can inhibit hyperplasia of the filtering passage tissue, helps to preserve the functional filtering blebs and prolong their life span, and induces apoptosis of RTFS.

Effect of aminolevulinic acid-based photodynamic therapy on expression of vascular endothelial growth factor and proliferating cell nuclear antigen in condyloma acuminatum tissues / 中华皮肤科杂志

Objective@#To evaluate the effect of aminolevulinic acid-based photodynamic therapy (ALA-PDT) on the expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in condyloma acuminatum (CA) tissues.@*Methods@#A total of 56 patients with CA were enrolled from Department of Dermatology, The First Affiliated Hospital of Zhengzhou University from October 2016 to September 2017, and skin lesions were obtained before and 1 week after the first ALA-PDT treatment. Immunohistochemical SP method was used to determine the expression of VEGF and PCNA in keratinocytes in the CA tissues. Chi-square test and rank sum test were carried out to analyze differences between pre- and post-treatment expression rate and intensity of VEGF and PCNA, and Spearman correlation analysis was conducted to analyze the correlations between the protein expression of VEGF and PCNA.@*Results@#The expression rates of VEGF and PCNA in keratinocytes in the CA tissues were 71.43% (40/56) and 73.21% (41/56) respectively before ALA-PDT, and 44.64% (25/56) and 41.07% (23/46) respectively after ALA-PDT. There were significant differences between pre- and post-treatment expression rate and intensity of VEGF and PCNA (expression rate: χ2 = 8.25, 11.81 respectively, both P < 0.05; expression intensity: H = 11.29, 12.22 respectively, both P < 0.05) . The expression of VEGF was positively correlated with the expression of PCNA in the CA tissues before and after the ALA-PDT treatment (rs = 0.202, 0.273, respectively, both P < 0.05) .@*Conclusion@#The expression of VEGF and PCNA decreased in CA tissues after ALA-PDT treatment, which may be one of the mechanisms underlying the treatment of CA with ALA-PDT.

Effect of aminolevulinic acid-based photodynamic therapy on expression of vascular endothelial growth factor and proliferating cell nuclear antigen in condyloma acuminatum tissues / 中华皮肤科杂志

Objective To evaluate the effect of aminolevulinic acid-based photodynamic therapy (ALA-PDT) on the expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in condyloma acuminatum (CA) tissues.Methods A total of 56 patients with CA were enrolled from Department of Dermatology,The First Affiliated Hospital of Zhengzhou University from October 2016 to September 2017,and skin lesions were obtained before and 1 week after the first ALA-PDT treatment.Immunohistochemical SP method was used to determine the expression of VEGF and PCNA in keratinocytes in the CA tissues.Chi-square test and rank sum test were carried out to analyze differences between pre-and post-treatment expression rate and intensity of VEGF and PCNA,and Spearman correlation analysis was conducted to analyze the correlations between the protein expression of VEGF and PCNA.Results The expression rates of VEGF and PCNA in keratinocytes in the CA tissues were 71.43％ (40/56) and 73.21％ (41/56) respectively before ALA-PDT,and 44.64％ (25/56) and 41.07％ (23/46)respectively after ALA-PDT.There were significant differences between pre-and post-treatment expression rate and intensity of VEGF and PCNA (expression rate:x2 =8.25,11.81 respectively,both P ＜ 0.05;expression intensity:H =11.29,12.22 respectively,both P ＜ 0.05).The expression of VEGF was positively correlated with the expression of PCNA in the CA tissues before and after the ALA-PDT treatment (rs =0.202,0.273,respectively,both P ＜ 0.05).Conclusion The expression of VEGF and PCNA decreased in CA tissues after ALA-PDT treatment,which may be one of the mechanisms underlying the treatment of CA with ALA-PDT.

Improved histoarchitectural changes with angiotensin receptor blockers in early testicular and cauda toxicity in rats / Mejora de los cambios en la histoarquitectura con bloqueadores de receptores de angiotensina en la toxicidad precoz testicular y de cauda en ratas

SUMMARY: Reproductive dysfunction is a complication for many diseases and toxins. Its early diagnosis and treatment are immensely important. Here the morphological histoarchitecture changes in early testicular and cauda toxicity before and after treatment with angiotensin receptor blockers were evaluated. Low-grade testicular damage was induced using thioacetamide (TAA, 50 mg/kg/day) intraperitoneally for two weeks in rats. The rats were randomly divided into four groups (n = 8) treated daily orally for three weeks as follows: Normal control (distilled water), TAA (positive control), TAA+candesartan (0.2 mg/kg) and TAA+losartan (7.5 mg/kg). Serum testosterone and testicular malondialdehyde and glutathione were measured. The changes in histoarchitecture of testis and cauda epididymis were evaluated by hematoxylin and eosin for general structure, Masson's trichrome for collagen, periodic acid Schiff for basement membrane, and caspase-3 and proliferating cell nuclear antigen (PCNA) for immunohistochemical analysis. The TAA-rats showed decreases of serum testosterone and testicular glutathione, increases in testicular malondialdehyde, degenerative changes and apoptosis in germ cells, thickening of tubular basal lamina and increases in expression of caspase 3, and decreases in expression of PCNA. The ARBs (candesartan and losartan) significantly reversed these changes with non-significant differences in-between. Treatment with ARBs (candesartan and losartan) significantly reversed TAA-induced low-grade testicular and cauda toxicity in rats. This could be potentially useful for early treatment of male patients with occupational toxicant-induced reproductive dysfunction especially if they are using ARBs for other comorbidities.

RESUMEN: La disfunción reproductiva es una complicación por muchas enfermedades y toxinas. Su diagnóstico y tratamiento tempranos son inmensamente importantes. Aquí se evaluaron los cambios morfológicos en la histoarquitectura en la toxicidad precoz testicular y cauda antes y después del tratamiento con bloqueadores de receptores de angiotensina. Se indujo daño testicular de bajo grado usando tioacetamida (TAA, 50 mg / kg / día) por vía intraperitoneal durante dos semanas en ratas. Las ratas se dividieron aleatoriamente en cuatro grupos (n = 8) tratados diariamente por vía oral durante tres semanas de la siguiente manera: control normal (agua destilada), TAA (control positivo), TAA + candesartan (0,2 mg / kg) y TAA + losartán (7,5 mg / kg). Se midieron la testosterona sérica, el malondialdehído testicular y el glutatión. Los cambios en la histoarquitectura de los testículos y la epidermis de la cauda se evaluaron mediante Hematoxilina y Eosina para determinar la estructura general, con tricrómicro de Masson para el colágeno, ácido periódico de Schiff para la membrana basal y la caspasa-3 y el antígeno nuclear de células proliferantes (PCNA) para análisis inmunohistoquímico. Las ratas TAA mostraron disminución de la testosterona sérica y glutatión testicular, aumentos en el malondialdehído testicular, cambios degenerativos y apoptosis en células germinales, engrosamiento de la lámina basal tubular y aumentos en la expresión de la caspasa 3, y disminución en la expresión de PCNA. Los ARB (candesartán y losartán) revirtieron significativamente estos cambios con diferencias no significativas en el medio. El tratamiento con BRA (candesartán y losartán) revirtió significativamente la toxicidad testicular y cauda inducida por TAA en ratas. Esto podría ser potencialmente útil para el tratamiento temprano de pacientes con disfunción reproductiva inducida por tóxicos ocupacionales, especialmente si están usando BRA para otras comorbilidades.