This research investigated the impact of Ganoderma lucidum polysaccharides(GLP) on hepatoblastoma HepG2 and Huh6 cell models, as well as KM mouse model with in situ transplanted tumors, so as to provide a theoretical basis for the clinical application of GLP. Cell viability was assessed through the CCK-8 assay, whereas cell proliferation was evaluated by using the BeyoClick~(TM)EdU-488 test. Cell apoptosis was visualized via Hochest 33258 staining, and autophagy was detected through Mrfp-GFP-LC3 dual fluorescence staining. An in situ tumor transplantation model was created by using HepG2 cells in mice, and mice were treated with normal saline and GLP of 100, 200, and 300 mg·kg~(-1) for tumor count calculation and size assessment. Hematoxylin-eosin(HE) staining was used to observe pathological changes in tumor tissue and vital organs(liver, kidney, lung, spleen, and heart). Western blot analysis was conducted to measure the protein expressions of tumor protein P53(P53), B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), cleaved-caspase-3, Beclin-1, autophagy related protein-5(Atg-5), microtubule-associated protein-light chain-3Ⅰ(LC3Ⅰ)/LC3Ⅱ, autophagy adapter protein 62(P62), protein kinase B(Akt), p-Akt, mammalian target of rapamycin(mTOR), and p-mTOR. The in vitro experiment revealed that compared with the control group, after GLP treatment, tumor cell viability decreased significantly; apoptosis rate increased in a dose-dependent manner, and autophagic flux was inhibited. The in vivo experiments showed that compared with the model group, mice treated with GLP exhibited significantly fewer and smaller tumors. Western blot results showed that compared with the control group or model group, levels of P53, Bax, cleaved-caspase-3, Beclin-1, Atg-5, and LC3-Ⅱ/LC3-Ⅰ were significantly increased after GLP treatment, and the levels of Bcl-2, P62, p-Akt/Akt, and p-mTOR/mTOR were significantly decreased. These outcomes suggest that GLP promotes apoptosis and autophagy in hepatoblastoma cells by regulating the Akt/mTOR pathway.
Animals ; Humans ; Autophagy/drug effects* ; Reishi/chemistry* ; Mice ; Apoptosis/drug effects* ; TOR Serine-Threonine Kinases/genetics* ; Proto-Oncogene Proteins c-akt/genetics* ; Liver Neoplasms/genetics* ; Hepatoblastoma/genetics* ; Polysaccharides/pharmacology* ; Cell Line, Tumor ; Signal Transduction/drug effects* ; Male ; Cell Proliferation/drug effects* ; Hep G2 Cells

Good endometrial receptivity is an essential factor for embryo implantation,and gene expression in endometrial tissue during the window of implantation(WOI)is closely related to receptivity.Transcriptome sequencing technology enables the identification of gene expression profiles of endometrium during different menstrual phases,as well as microRNAs and long-chain non-coding RNA sequences involved in regulating gene expression.Combining this technology with bioinformatics analysis provides a better understanding of specific gene expression during the receptive period and offers technical support for studying its regulatory mechanism.Moreover,gene expression profiles of the endometrium during different menstrual phases hold significant clinical application value for accurately assessing endometrium receptivity in infertility patients and those with repeated implantation failure,thereby guiding individualized embryo transfer strategies.This review summarizes the progress of transcriptome sequencing in evaluating human endometrial receptivity and discusses future research directions.This review aims to understand the complex molecular mechanisms of endometrial receptivity formation and regulation from the transcriptional level,in order to improve the implantation rate of embryos in assisted reproductive technology and reduce the abortion rate.

AIM To study the chemical constituents from the stems and leaves of Dendrobium formosum Roxb.ex Lindl.and their biological activities.METHODS The 95％ethanol extract from the stems and leaves of D.formosum was isolated and purified by silica gel,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.Their inhibitory activities onα-glucosidase were determined by PNPG method,and their in vitro anti-inflammatory activities were evaluated by RAW264.7 model.RESULTS Fifteen compounds were isolated and identified as coniferyl p-coumarate(1),(-)-pinoresinol(2),2,5,7-trihydroxy-4-methoxy-9,10-dihydrophenanthrene(3),naringenin(4),spiropreussomerin A(5),7-hydroxy-14-de-O-methyl-lasiodiplodin(6),(4S,5S,6Z,8E)-5-hydroxydeca-6,8-dien-4-olide(7),(6S,9R)-blumenol C(8),p-hydroxybenzoic acid(9),m-hydroxybenzoic acid(10),p-hydroxy benzenepropanoic acid(11),5,7-dihydroxy-isobenzofuran(12),2-(4-hydroxyphenyl)-ethanol(13),β-sitostenone(14),β-sitosterol(15).The IC50 values of compounds 1 and 4 on α-glucosidase inhibition were(65.60±3.31)and(98.95±2.53)μmol/L,respectively.Compound 3 presented inhibitory activity on NO production in RAW 264.7 cells,with IC50 value of(3.97±0.12)μmol/L.CONCLUSION Compounds 5-6,8 and 12 are isolated from Orchidacae family for the first time,and 2-15 are first isolated from this plant.Compounds 1 and 4 have α-glucosidase inhibitory activities,and 3 has anti-inflammatory activity.

AIM To study the chemical constituents from the stems and leaves of Dendrobium formosum Roxb.ex Lindl.and their biological activities.METHODS The 95％ethanol extract from the stems and leaves of D.formosum was isolated and purified by silica gel,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.Their inhibitory activities onα-glucosidase were determined by PNPG method,and their in vitro anti-inflammatory activities were evaluated by RAW264.7 model.RESULTS Fifteen compounds were isolated and identified as coniferyl p-coumarate(1),(-)-pinoresinol(2),2,5,7-trihydroxy-4-methoxy-9,10-dihydrophenanthrene(3),naringenin(4),spiropreussomerin A(5),7-hydroxy-14-de-O-methyl-lasiodiplodin(6),(4S,5S,6Z,8E)-5-hydroxydeca-6,8-dien-4-olide(7),(6S,9R)-blumenol C(8),p-hydroxybenzoic acid(9),m-hydroxybenzoic acid(10),p-hydroxy benzenepropanoic acid(11),5,7-dihydroxy-isobenzofuran(12),2-(4-hydroxyphenyl)-ethanol(13),β-sitostenone(14),β-sitosterol(15).The IC50 values of compounds 1 and 4 on α-glucosidase inhibition were(65.60±3.31)and(98.95±2.53)μmol/L,respectively.Compound 3 presented inhibitory activity on NO production in RAW 264.7 cells,with IC50 value of(3.97±0.12)μmol/L.CONCLUSION Compounds 5-6,8 and 12 are isolated from Orchidacae family for the first time,and 2-15 are first isolated from this plant.Compounds 1 and 4 have α-glucosidase inhibitory activities,and 3 has anti-inflammatory activity.

Objective To investigate the curative effect of early hemoperfusion(HP)in treating patients with sepsis.Methods Early stage sepsis patients admitted to the First People's Hospital of Nanyang City Affiliated to Henan University from January 2020 to December 2022 were selected as the research subjects.These patients were randomly divided into experimental group(62 patients)and control group(31 patients).Early bundle therapy combined with continuous renal replacement therapy(CRRT)were used in both groups as the standard treatment,but only the standard treatment was used in control group.Based on the treatment of control group,the patients in experimental group were treated with HP,the"2-2-1"plan(on the first and second days,connecting resin HP device HA380 in series every 12 hours,and connecting one perfusion device in series on the third day)was applied.The white blood cell count(WBC),C-reactive protein(CRP),procalcitonin(PCT),tumor necrosis factor-α(TNF-α),interleukins(IL-6,IL-10),the hemodynamic indicators(systolic blood pressure,diastolic blood pressure),lactic acid(Lac),sequential organ failure assessment(SOFA),acute physiology and chronic health evaluationⅡ(APACHEⅡ),and the difference in intensive care unit(ICU)hospitalization time of the patients before admission and after 3 day treatment were compared between the two groups.The Kaplan-Meier method was used to draw survival curves and the difference in 28-day survival rates was compared between the two groups.Results After 3 days of treatment,the inflammatory indexes TNF-α,IL-6,IL-10,PCT,CRP,WBC and SOFA score,APACHEⅡscore,Lac in the experimental group all decreased significantly(all P<0.05).In the control group,only the PCT and CRP indexes significantly decreased compared with those before treatment(both P<0.05).Compared with the control group,the difference before and after the change of PCT,CRP,IL-6,IL-10 and SOFA score,APACHEⅡscore,Lac in the experimental group were significant[PCT(μg/L):12.31(6.95,42.69)vs.3.84(1.06,14.47),CRP(mg/L):78.69±64.60 vs.31.49±81.14,IL-6(ng/L):1 357.20(243.96,7 205.50)vs.110.00(-1 749.00,1 377.00),IL-10(ng/L):36.87(5.43,1 218.80)vs.2.67(-11.00,22.79),SOFA score:5(3,7)vs.1(-3,6),APACHEⅡscore:8(5,11)vs.5(-2,7),Lac(mmol/L):3.93±2.89 vs.2.42±2.89,all P<0.05].The clearance rate of inflammatory cytokines IL-6 and IL-10 in the experimental group was higher than that in the control group(IL-6:F=17.93,P<0.001,IL-10:F=6.62,P=0.012).The Kaplan-Meier curve showed the 28-day survival rate of the experimental group was higher than that of the control group(79.03%vs.58.06%),the differences were statistically significant(Log-Rank test:χ2=4.706,P=0.030).Multivariate Cox regression analysis showed that no HP treatment,Lac level before treatment and high APACHEⅡ score were risk factors for death Conclusion HP in the early interventional treatment of sepsis can reduce the levels of inflammatory cytokines and control the inflammatory storm.The HP"2-2-1"treatment frequency can effectively improve the prognosis of septic patients and increase their survival rate.

Objective To investigate the curative effect of early hemoperfusion(HP)in treating patients with sepsis.Methods Early stage sepsis patients admitted to the First People's Hospital of Nanyang City Affiliated to Henan University from January 2020 to December 2022 were selected as the research subjects.These patients were randomly divided into experimental group(62 patients)and control group(31 patients).Early bundle therapy combined with continuous renal replacement therapy(CRRT)were used in both groups as the standard treatment,but only the standard treatment was used in control group.Based on the treatment of control group,the patients in experimental group were treated with HP,the"2-2-1"plan(on the first and second days,connecting resin HP device HA380 in series every 12 hours,and connecting one perfusion device in series on the third day)was applied.The white blood cell count(WBC),C-reactive protein(CRP),procalcitonin(PCT),tumor necrosis factor-α(TNF-α),interleukins(IL-6,IL-10),the hemodynamic indicators(systolic blood pressure,diastolic blood pressure),lactic acid(Lac),sequential organ failure assessment(SOFA),acute physiology and chronic health evaluationⅡ(APACHEⅡ),and the difference in intensive care unit(ICU)hospitalization time of the patients before admission and after 3 day treatment were compared between the two groups.The Kaplan-Meier method was used to draw survival curves and the difference in 28-day survival rates was compared between the two groups.Results After 3 days of treatment,the inflammatory indexes TNF-α,IL-6,IL-10,PCT,CRP,WBC and SOFA score,APACHEⅡscore,Lac in the experimental group all decreased significantly(all P<0.05).In the control group,only the PCT and CRP indexes significantly decreased compared with those before treatment(both P<0.05).Compared with the control group,the difference before and after the change of PCT,CRP,IL-6,IL-10 and SOFA score,APACHEⅡscore,Lac in the experimental group were significant[PCT(μg/L):12.31(6.95,42.69)vs.3.84(1.06,14.47),CRP(mg/L):78.69±64.60 vs.31.49±81.14,IL-6(ng/L):1 357.20(243.96,7 205.50)vs.110.00(-1 749.00,1 377.00),IL-10(ng/L):36.87(5.43,1 218.80)vs.2.67(-11.00,22.79),SOFA score:5(3,7)vs.1(-3,6),APACHEⅡscore:8(5,11)vs.5(-2,7),Lac(mmol/L):3.93±2.89 vs.2.42±2.89,all P<0.05].The clearance rate of inflammatory cytokines IL-6 and IL-10 in the experimental group was higher than that in the control group(IL-6:F=17.93,P<0.001,IL-10:F=6.62,P=0.012).The Kaplan-Meier curve showed the 28-day survival rate of the experimental group was higher than that of the control group(79.03%vs.58.06%),the differences were statistically significant(Log-Rank test:χ2=4.706,P=0.030).Multivariate Cox regression analysis showed that no HP treatment,Lac level before treatment and high APACHEⅡ score were risk factors for death Conclusion HP in the early interventional treatment of sepsis can reduce the levels of inflammatory cytokines and control the inflammatory storm.The HP"2-2-1"treatment frequency can effectively improve the prognosis of septic patients and increase their survival rate.

Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.

Objective: This cross-sectional investigation aimed to determine the incidence, clinical characteristics, prognosis, and related risk factors of olfactory and gustatory dysfunctions related to infection with the SARS-CoV-2 Omicron strain in mainland China. Methods: Data of patients with SARS-CoV-2 from December 28, 2022, to February 21, 2023, were collected through online and offline questionnaires from 45 tertiary hospitals and one center for disease control and prevention in mainland China. The questionnaire included demographic information, previous health history, smoking and alcohol drinking, SARS-CoV-2 vaccination, olfactory and gustatory function before and after infection, other symptoms after infection, as well as the duration and improvement of olfactory and gustatory dysfunction. The self-reported olfactory and gustatory functions of patients were evaluated using the Olfactory VAS scale and Gustatory VAS scale. Results: A total of 35 566 valid questionnaires were obtained, revealing a high incidence of olfactory and taste dysfunctions related to infection with the SARS-CoV-2 Omicron strain (67.75%). Females(χ2=367.013, P<0.001) and young people(χ2=120.210, P<0.001) were more likely to develop these dysfunctions. Gender(OR=1.564, 95%CI: 1.487-1.645), SARS-CoV-2 vaccination status (OR=1.334, 95%CI: 1.164-1.530), oral health status (OR=0.881, 95%CI: 0.839-0.926), smoking history (OR=1.152, 95%CI=1.080-1.229), and drinking history (OR=0.854, 95%CI: 0.785-0.928) were correlated with the occurrence of olfactory and taste dysfunctions related to SARS-CoV-2(above P<0.001). 44.62% (4 391/9 840) of the patients who had not recovered their sense of smell and taste also suffered from nasal congestion, runny nose, and 32.62% (3 210/9 840) suffered from dry mouth and sore throat. The improvement of olfactory and taste functions was correlated with the persistence of accompanying symptoms(χ2=10.873, P=0.001). The average score of olfactory and taste VAS scale was 8.41 and 8.51 respectively before SARS-CoV-2 infection, but decreased to3.69 and 4.29 respectively after SARS-CoV-2 infection, and recovered to 5.83and 6.55 respectively at the time of the survey. The median duration of olfactory and gustatory dysfunctions was 15 days and 12 days, respectively, with 0.5% (121/24 096) of patients experiencing these dysfunctions for more than 28 days. The overall self-reported improvement rate of smell and taste dysfunctions was 59.16% (14 256/24 096). Gender(OR=0.893, 95%CI: 0.839-0.951), SARS-CoV-2 vaccination status (OR=1.334, 95%CI: 1.164-1.530), history of head and facial trauma(OR=1.180, 95%CI: 1.036-1.344, P=0.013), nose (OR=1.104, 95%CI: 1.042-1.171, P=0.001) and oral (OR=1.162, 95%CI: 1.096-1.233) health status, smoking history(OR=0.765, 95%CI: 0.709-0.825), and the persistence of accompanying symptoms (OR=0.359, 95%CI: 0.332-0.388) were correlated with the recovery of olfactory and taste dysfunctions related to SARS-CoV-2 (above P<0.001 except for the indicated values). Conclusion: The incidence of olfactory and taste dysfunctions related to infection with the SARS-CoV-2 Omicron strain is high in mainland China, with females and young people more likely to develop these dysfunctions. Active and effective intervention measures may be required for cases that persist for a long time. The recovery of olfactory and taste functions is influenced by several factors, including gender, SARS-CoV-2 vaccination status, history of head and facial trauma, nasal and oral health status, smoking history, and persistence of accompanying symptoms.
Female ; Humans ; Adolescent ; SARS-CoV-2 ; Smell ; COVID-19/complications* ; Cross-Sectional Studies ; COVID-19 Vaccines ; Incidence ; Olfaction Disorders/etiology* ; Taste Disorders/etiology* ; Prognosis

In the perspective of technical evaluation, the pre-marketing regulatory requirements of allergen detection reagents in China, America, European Union were compared, and the regulatory risks and performance requirements of this product were analyzed based on the monitoring of post-marketing adverse events, reference standards and domestic and foreign regulatory documents. In view of the "neck-stuck" problems such as the difficulty of clinical trials, the difficulty of finding comparable contrast reagents and the lack of clinical diagnostic gold standards, this paper discusses and gives regulatory suggestions, with a view to providing technical reference for product R&D, production, evaluation, approval and supervision in this field.
Allergens ; European Union ; Indicators and Reagents ; Marketing ; Reference Standards

Aim To observe the inhibitory effect of neferine(Nef)on the migration and invasion of non-small cell lung cancer(NSCLC)H1299 cells by blocking ROCK pathway.Methods H1299 cells were taken for in vitro culture, and treated with different concentrations of Nef.H1299 cell viability was measured by CCK-8 method to determine the dose of the experimental group.The migration and invasion abilities of H1299 cells were detected by cell scratch test and Transwell chamber test.The expression of matrix metalloproteinases MMP-2 and MMP-9 secreted from lung cancer cells was detected by enzyme linked immunosorbent assay(ELISA).The protein level of ROCK1 in H1299 cells was tested by real-time fluorescent quantitative PCR and Western blot; the binding mode and affinity between Nef and ROCK1 were stimulated by AutoDock semi flexible docking method.Results The doses of Nef in the experimental group were determined as 4, 6 and 10 μmol·L-1.These three concentrations of Nef could inhibit the migration and invasion of H1299 lung cancer cells to a certain degree in a dose-dependent manner.At the same time, Nef reduced the expression of MMP-2, MMP-9 and ROCK1 proteins related to the migration and invasion of the cancer cells.In addition, the affinity of Nef to ROCK1 was significantly higher than that of fasudil, an inhibitor of ROCK, and the binding force was stronger to A-chain of ROCK1.Conclusions As a potential natural anticancer compound, Nef can inhibit the migration and invasion of NSCLC by reducing the expression of MMP-2, MMP-9 and ROCK1 proteins related to the migration and invasion of the cancer cells.