Objective To explore the effect of microRNA-32 (miRNA-32)on the biological behaviors of gastric cancer cell and its mechanism.Methods Gastric cancer cell line SGC-7901 cells were transiently transfected with miRNA-32 analogue,miRNA-32 inhibitor and empty plasmid vectors by lipofectamine and divided into analogue transfection group,inhibitor transfection group,empty plasmid transfection group and non-transfection group.The expression of green fluorescent protein was observed under fluorescent microscopy.The expression of miRNA-32 at mRNA level was detected by quantificational real-time polymerase chain reaction.The cell proliferation was evaluated by CCK-8 assay.The cell migration ability was measured by scratch test and Transwell chamber assays.The data were analyzed by one-way ANOVA.Results Compared with empty plasmid transfection group and non-transfection group,the expression of miRNA-32 mRNA of miRNA-32 analogue transfection group (relative quantitative value:2.327) was significantly up-regulated and that of miRNA-32 inhibitor transfection group (relative quantitative value:0.402) was significantly down regulated (F=11.238,P＜0.05).The width of scratch of miRNA-32 analogue transfection group was (61.39± 2.21) μm at 24 hours; miRNA-32 inhibitor transfection group was (29.97±0.66) μm.The migration distance of inhibitor transfection group was far than that of analogue transfection group (F=9.371,P＜0.05).After transfection for 48 hours,the cell number of migrated cells of analogue transfection group was significantly less than that of non transfection group,which was 16.93±4.63 and 93.93± 7.09,respectively (F=6.853,P＜0.05).After transfection for 48 hours and 72 hours,the cell growth inhibiting rate of miRNA 32 analogue transfection group was (43.474 ± 18.636)％ and (45.050±23.764)％,respectively,the cell growth was significantly inhibited (F=7.986 and 8.635,P=0.028 and 0.032).Conclusion The cell growth and migration ability of human gastric cancer cell line SGC-7901 are obviously inhibited through upregulating the expression of miRNA-32.

OBJECTIVETo construct a recombinant prokaryotic expression vector (plasmid) containing microneme protein 1 (MIC1) partial gene in toxoplasma gondii (T. gondii) ZS2 isolate. The gene was expressed in varied Escherichia coli (E. coli) after sequencing.

METHODSThe gene fragment coding MIC 1 from the genomic DNA of T. gondii ZS2 isolate was amplified by polymerase chain reaction (PCR). The gene was inserted to a prokaryotic expression vector pWR450-1 by digesting with restriction enzymes and linking reaction. The positive clone was screened on LB plates containing ampicillin and identified by restrictive enzyme digestion, PCR amplification and sequence analysis. The recombinant plasmid was transferred into E. coli TG1, JM109 (DE3) and DH5 alpha, and was expressed under the induction of IPTG. The expression products were identified by SDS-PAGE. The MIC1 gene structure was analyzed and compared in homology with the gene sequence of RH isolate using computer software.

RESULTSThe recombinant plasmid pWR450-1/MIC1, after cloning from acquired 471 bp MIC1 gene fragment and amplified from the genome gene ZS2, was complete homologous to the sequence of RH isolate, reflecting its highly conservative. The gene could be expressed as fusion protein with 70,000 in varied E. coli.

CONCLUSIONRecombinant plasmid pWR450-MIC1 was successfully constructed and could be expressed in different strains of E. coli, laying a foundation for research on its structure and function.

Animals ; Base Sequence ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Cloning, Molecular ; DNA, Protozoan ; analysis ; Escherichia coli ; genetics ; Gene Expression ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques ; Protozoan Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Toxoplasma ; genetics

OBJECTIVETo investigate the protective effects of oxysophoridine (OSR) on primary cultured hippocampus neurons subjected to anoxia injury in neonatal rats and its mechanism.

METHODThe model of anoxia injury of hippocampus neurons in neonatal rats were primarily cultured in vitro by physical oxygen deficiency using glucose-free culture fluid. The survival rate of neurons, the leaking rate of lactate dehydrogenase (LDH), the intracellular contents of malondialdehyde (MDA) and nitric oxide (NO), the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and nitric oxide synthase (NOS) were measured. The intracellular free calcium concentration ([Ca2+]i) in hippocampus neurons were detected with Ca(2+)-sensitive dual wavelength fluorescence spectrophotometer.

RESULTNeuron death occurred in the anoxia injury model group with increase of LDH leaking rate, the contents of NO, MDA, intracellular [Ca2+] and the elevated activity of NOS while decreased activities of SOD and GSH-PX. The hippocampus neurons subjected to anoxia injury were alleviated in OSR (0.625, 5, 10 microg x L(-1)) group.

CONCLUSIONOSR has significant protective effects on hippocampus neurons subjected to anoxic injury. The mechanism of its protective effect may relate to its reduction of calcium overload and against oxidation injury.

Alkaloids ; administration & dosage ; Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Glutathione Peroxidase ; metabolism ; Hippocampus ; cytology ; drug effects ; enzymology ; metabolism ; Humans ; Hypoxia ; drug therapy ; enzymology ; metabolism ; prevention & control ; Malondialdehyde ; metabolism ; Neurons ; cytology ; drug effects ; enzymology ; metabolism ; Nitric Oxide Synthase ; metabolism ; Protective Agents ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Sophora ; chemistry ; Superoxide Dismutase ; metabolism

Alzheimer's disease (AD) is the general neurodegenerative disease in the elderly. Alterations of microtubule associated protein tau are closely related to the onset and progression of AD. The role of tau protein in the development of AD has been a focus in the current study of AD, with an emphasis on the identification of compounds that inhibit tau protein phosphorylation and aggregate, and accelerate tau protein depolymerization. The current research status of tau protein as target of AD preventive and therapeutic drug was also reviewed.

OBJECTIVESix1 and Six4 are expressed in several tumors, and associated with tumor progress and poor prognosis. The aim of this study was to investigate the expression of Six1 and Six4 in esophageal squamous cell carcinoma (ESCC), and to evaluate their correlation with the clinicopathological factors and prognosis.

METHODSTissue microarray technology and immunohistochemical method (EnVision) were used to detect the expression of Six1 and Six4 in the tumor tissues and corresponding adjacent normal epithelium of esophagus from 292 ESCC patients.

RESULTSAmong the 292 ESCC patients, the positive rates of Six1 and Six4 protein expression in tumor tissues were 72.9% (213/292) and 56.2% (164/292), respectively, significantly higher than the expression rate of 33.2% (97/292) and 32.5% (95/292) in adjacent normal epithelium of esophagus (P < 0.05). Chi square test showed that the expression of Six1 protein was related to tumor size, depth of tumor invasion and patient survival status; higher Six4 protein expression level was related to poor differentiation and increased depth of invasion. Single factor Log-rank analysis revealed that gender, TNM stage, Six1 protein expression level were related to the overall survival of ESCC patients (P < 0.05), while the five-year survival rate was significantly higher in the Six1-negative group than the Six1-positive group [51.9% (41/79) vs. 43.7% (93/213)]. Multi-factor Cox proportional risk model analysis showed that TNM stage and positive expression of Six1 were independent prognostic factors for ESCC patients (P < 0.05).

CONCLUSIONSSix1 and Six4 are highly expressed in ESCC. Their expression levels are closely related to the progress and prognosis of ESCC. Over-expression of Six1 is related to poor prognosis in ESCC patients. Thus, Six1 could be used as an important prognostic indicator for ESCC patients.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Esophageal Neoplasms ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Homeodomain Proteins ; metabolism ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Risk Factors ; Survival Rate ; Trans-Activators ; metabolism ; Tumor Burden

Objective To analyze the effects of anesthesia-induced thermoregulatory system impairment and low temperature environment of the operating room on the perioperative thermoregulation of individualized patients by constructing a computer simulation model. Methods A simple anesthesia model was proposed and then incorporated into the self-developed individualized thermoregulatory model, in which human body was represented as a cylinder with two layers of the core and the skin. The integrated model could be used to assess the effects of individualized characteristics such as age, obesity, and cardiovascular diseases on thermoregulation by modifying different physiological parameters involving sweating, shivering and cutaneous vasomotion. Simulation of the general anesthesia effects on human thermoregulation could be achieved by reducing basal metabolic rate and thresholds for vasoconstriction and shivering. Results The elderly people showed lower core temperature but higher skin temperature, compared with the young people. In a low temperature environment, an increase in fat thickness or an increase in severity degree of the left ventricular failure (LVF) might alleviate the decrease in core temperature, while an increase in wind speed or relative humidity could result in a decrease in core temperature. When the threshold setting of vasoconstriction was reduced by 0-5-3 ℃, the core temperature showed a significant decrease. Conclusions By comparing model simulations with experimental measurements, the reliability and validity of the model in predicting human transient thermal responses during varying external thermal environment was verified. The individualized characteristics of human body had an important influence on human body temperature in a low temperature environment. Moreover, the combination of individualized characteristics of human body and general anesthesia further complicated the body′s thermoregulation and posed significant challenges for clinicians.

TAM(tumor-associated macrophage)is a kind of immune cell in tumor microenvironment,which mainly exists in tumor matrix to mediate inflammatory reaction.Generally,TAM can be stimulated by different cytokines to polarize into M1 macrophage or M2 macrophage with different phenotypes.In the early stage of tumor,M1 macrophages in TAM exhibit pro-inflammatory and anti-tumor activities,while as tumor progresses,TAMs tend to polarize into M2 macrophages to play anti-inflammatory and tumor-promoting roles.A large number of studies have shown that TAM is closely related to tumor growth,invasion,metastasis and poor prognosis.Therefore,targeting TAM has become the focus of anti-tumor immunotherapy.This paper has summarized the origin of TAM,and introduced the specific roles of TAM in promoting tumor proliferation,angiogenesis,migration and invasion and the formation of immunosuppressive environment.At the same time,the research progress of TAM-targeted anti-tumor immu-notherapy in recent years was discussed from 3 aspects:inhibiting monocyte recruitment,promoting TAM apoptosis and reshaping TAM phenotype.

By using computer-aided drug design, the activities group model which CDK4/6 inhibitors on the market were introduced to silybin C-7, and a series of silybin derivatives were designed and synthesized, and the structure was confirmed by MS, ¹³C NMR and ¹H NMR. The in vitro antitumor activity evaluation of the target compound was carried out by MTT method, and the in vitro anti-tumor activity was carried out in human hepatocellular carcinoma cells (HepG-2). Experimental results show that all compounds are higher than the activity of the parent silybin, of which compound I₁ has a certain inhibitory effect on human HepG-2 cells, which is worth further study.

Objective To systematicly evaluate the constituent factors and accuracy of prediction models of outcome for patients with prolonged disorders of consciousness. Methods Articles about prediction models of outcome for patients with prolonged disorders of consciousness were retrieved from PubMed, Web of Science, CNKI and Wanfang Data until September 30th, 2021. The authors, publishing times, subjects, predictive indicators, outcomes and conclusions were extracted. Results A total of 4 313 articles were returned and 37 included, comprising randomized controlled trials and cohort studies, which published mainly from 2012 to 2021. The subjects were patients with prolonged disorders of consciousness, and their predictions consisted of clinical assessment scales, neuroimaging, neuroelectrophysiology and laboratory indicators. Conclusion Prediction models may be valuable for the long-term outcomes of patients with prolonged disorders of consciousness. Most of the current prediction models are composed of only a limited number of technical means, and the accuracy is uneven. Coma Recovery Scale-revised, default mode network and multiple evoked potentials-related prediction models are accurate, but lack a unified adaptation standard.

Digestive tract diseases， especially digestive tract tumors， including liver cancer， pancreatic cancer， and colorectal cancer， have high incidence in China. Digestive tract tumor is one of the top 10 cancers in terms of the number of new cases and deaths in the world， and the incidence and mortality of tumor diseases have been increasing year by year. Therefore， the prevention and treatment of tumors is particularly important. With the application and promotion of traditional Chinese medicine in the medical field and the rapid development of molecular biology and pharmacology， more and more potential active components of Chinese medicinal materials have been extracted and studied. These active components can inhibit tumor cells in a multi-target and multi-pathway manner. Cinobufotalin is an effective component extracted from the skin of Bufo bufo gargarizans. It has been prepared into a variety of agents with anti-tumor， immunomodulatory， cardiac boosting， pain-easing， anti-inflammatory， and swelling-relieving activities. In clinical practice， cinobufotalin is mainly used to assist the treatment of liver cancer， lung cancer， colorectal cancer， gastric cancer and other malignant tumors， which can reduce the adverse reactions of patients in the middle and late stages and improve the quality of life and five-year survival rate of patients. The available studies of molecular mechanism have demonstrated that cinobufotalin can play a therapeutic role by inducing cell apoptosis， regulating cell cycle， inhibiting cell proliferation and angiogenesis， modulating immune response， reversing multidrug resistance， enhancing radiochemotherapy sensitivity， inhibiting tumor inflammation， invasion， and metastasis， etc. This review focuses on the clinical application and mechanism of cinobufotalin against digestive tract tumors in recent years， aiming to provide a theoretical basis for the anti-tumor research of cinobufotalin， promote the application of cinobufotalin in tumor treatment， and facilitate the further research and development of this compound.