ObjectiveTo investigate the chemosensitization effect of gallic acid （GA） combined with sorafenib （Sora） on hepatocellular carcinoma HepG2 cells and related mechanisms. MethodsHepG2 cells were randomly divided into control group， GA group， Sora group， and GA+Sora group. CCK8 assay was used to measure cell viability； CompuSyn software was used to analyze combination index （CI）； colony formation assay was used to evaluate the colony formation ability of cells； flow cytometry was used to measure cell apoptosis； wound healing assay and Transwell chamber assay were used to observe the migration and invasion abilities of cells； Western Blot was used to measure the expression matrix metalloproteinase 2 （MMP-2）， matrix metalloproteinase 9 （MMP-9）， and apoptosis-related proteins. HepG2 cells were subcutaneously inoculated into the lower right back of mice， and 6 days later， the mice were divided into control group， GA group， Sora group， and GA+Sora group. Tumor size and body weight were measured once a week， and drug intervention was performed for 21 days. Then the nude mice were sacrificed， and tumor weight was measured. 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. ResultsThe mean IC₅₀ values of GA and Sora for the treatment of HepG2 cells for 48 hours were 123.47±5.16 μmol/L and 9.87±0.98 μmol/L， respectively， and when Sora was combined with 70 μmol/L GA （IC₃₀）， IC₅₀ decreased to 2.06±0.35 μmol/L； the CI value was<1 for Sora at different concentrations combined with 70 μmol/L GA. The number of cell colonies was 234.0±20.4， 147.0±12.1， 129.3±13.3， and 73.0±7.6， respectively， in the four groups， and the GA+Sora group had a significantly lower number of cell colonies than the control group， the GA group， and the Sora group （all P<0.05）. After 48 hours of treatment， the cell apoptosis rate was 1.98%±0.29%， 15.17%± 1.56%， 18.65%±1.48%， and 34.60%±5.36%， respectively， in the four groups， and the GA+Sora group had a significantly higher cell apoptosis rate than the control group， the GA group， and the Sora group （all P<0.05）. After 24 hours of treatment， the cell migration rate was 55.59%±5.08%， 29.34%±4.36%， 21.80%±5.16%， and 6.47%±2.75%， respectively， in the four groups， and the GA+Sora group had a significantly lower cell migration rate than the control group， the GA group， and the Sora group （all P<0.05）. After 48 hours of treatment， the number of transmembrane cells was 223.7±13.0， 168.3±10.9， 155.3±29.1， and 62.7±19.7， respectively， in the four groups， and the GA+Sora group had a significantly lower number of transmembrane cells than the control group， the GA group， and the Sora group （all P<0.05）. Compared with the control group， the GA group， the Sora group， and the GA+Sora group had significant reductions in the protein expression levels of MMP-2， MMP-9， and Bcl-2 （all P<0.05） and significant increases in the protein expression levels of Bax and cleaved caspase-3 （all P<0.05）. Compared with the control group， the GA， Sora， and GA+Sora groups had significant reductions in tumor volume and weight （all P<0.05）， and compared with the Sora group， the GA+Sora group had significant reductions in tumor volume and weight in nude mice （both P<0.05）. ConclusionGA can increase the sensitivity of HepG2 cells to Sora chemotherapy， possibly by promoting cell apoptosis and inhibiting cell migration and invasion after combination with Sora.

ObjectiveTo investigate the effect of gallic acid （GA） on the proliferation， migration， invasion， and apoptosis of human hepatocellular carcinoma HepG2 cells and its mechanism. MethodsHepG2 cells were treated with different concentrations of GA （0， 5， 10， 20， 30， 40， and 50 μg/mL） for 24 and 48 hours， and CCK8 assay was used to measure cell viability and calculate IC₅₀. The experiment was divided into control group （HepG2 cells）， 5 μg/mL GA group， 10 μg/mL GA group， and 20 μg/mL GA group. Plate colony formation assay was used to evaluate the effect of GA on cell proliferation； wound healing assay and Transwell chamber assay were used to observe the effect of GA on cell migration and invasion； flow cytometry was used to observe the effect of GA on cell apoptosis； Western blot was used to measure the expression of matrix metallopeptidase-2 （MMP-2）， matrix metallopeptidase-9 （MMP-9）， and apoptosis-related proteins. 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. ResultsThe mean IC₅₀ value of GA on HepG2 cells was 38.02±2.58 μg/mL at 24 hours and 18.36±1.54 μg/mL at 48 hours. The number of cell colonies was 239.00±29.45 in the control group， 210.00±19.00 in the 5 μg/mL GA group， 144.33±16.03 in the 10 μg/mL GA group， and 57.00±9.55 in the 20 μg/mL GA group， suggesting that compared with the control group， each GA group had a significant reduction in cell colony formation ability （all P<0.05）. After 24 hours of treatment， the cell migration rate was 42.62%± 7.82% in the control group， 35.34%±6.42% in the 5 μg/mL GA group， 21.85%±4.42% in the 10 μg/mL GA group， and 12.57%± 3.54% in the 20 μg/mL GA group， respectively， in these four groups， and the number of transmembrane cells in these four groups was 230.30±15.30， 182.12±12.60， 137.20±7.50， and 124.40±6.80， respectively， suggesting that compared with the control group， each GA group had significant reductions in migration rate and the number of transmembrane cells （all P<0.05）. After 48 hours of treatment， the cell apoptotic rate was 0.67%±0.08% in the control group， 13.27%±1.07% in the 5 μg/mL GA group， 20.94%± 2.45% in the 10 μg/mL GA group， and 40.74%±2.63% in the 20 μg/mL GA group， and compared with the control group， each GA group had a significant increase in cell apoptosis rate （all P<0.05）. Compared with the control group， each GA group had significant reductions in the protein expression levels of MMP-2 and MMP-9 （all P<0.05） and significant increases in the protein expression levels of Bax and cleaved caspase-3 （all P<0.05）. ConclusionGA can inhibit the proliferation， migration， and invasion of HepG2 cells and promote the apoptosis of HepG2 cells， possibly by regulating MMP-2， MMP-9， and the apoptosis-related proteins Bax/Bcl-2.

ObjectiveTo evaluate the efficacy of Shexiang Baoxinwan in treating stable angina pectoris with Qi stagnation and blood stasis syndrome in patients with coronary artery disease （CAD） complicated with anxiety and depression and explore its underlying mechanisms. MethodsThis study employed a randomized， double-blind， and placebo-controlled clinical trial design. Patients admitted to the hospital were randomly assigned to the observation group and the control group， with 52 patients in each group. Patients in the observation and control groups received Shexiang Baoxinwan and placebo， respectively， both in combination with conventional Western medication. The dose was 45.0 mg， three times daily， for a total duration of eight weeks. The primary outcome was the Seattle Angina Questionnaire （SAQ） scores before and after treatment. Secondary outcomes included changes in traditional Chinese medicine （TCM） syndrome score， the patient health questionnaire-9 （PHQ-9）， generalized anxiety disorder-7 （GAD-7）， inflammatory markers ［interleukin-18 （IL-18）， interleukin-10 （IL-10）， tumor necrosis factor-alpha （TNF-α）， CD40， etc.］， monoamine neurotransmitters ［e.g.， dopamine （DA）］， vascular endothelial function markers ［e.g.， endothelin-1（ET-1）］， adipokines， and ischemia-modified albumin （IMA）. Adverse reactions were also recorded. ResultsA total of 92 patients completed the study， with 44 in the observation group and 48 in the control group. Compared with baseline， both groups showed significant decreases in PHQ-9， GAD-7， and TCM syndrome scores following treatment （P<0.05）， along with a significant increase in SAQ scores （P<0.05）. In the observation group， DA levels were significantly increased （P<0.05）， while levels of IL-18， TNF-α， CD40， ET-1， and IMA were decreased （P<0.05）. In contrast， the control group exhibited significantly increased CD40 levels （P<0.05）. Compared with the control group after treatment， the observation group showed significant improvements in the SAQ dimensions of physical limitation， angina stability， treatment satisfaction， and disease perception， as well as in TCM syndrome score， PHQ-9 score， IL-18， CD40， ET-1， and IMA （P<0.05）. No adverse reactions were observed in either group during treatment. ConclusionShexiang Baoxinwan can improve anxiety and depression， alleviate angina symptoms， and reduce TCM symptoms of Qi stagnation and blood stasis in CAD patients. The mechanism may involve anti-inflammation， improvement of vascular endothelial function， reduction of IMA， and increase of monoamine neurotransmitter levels.

ObjectiveTo evaluate the efficacy of Shexiang Baoxinwan in treating stable angina pectoris with Qi stagnation and blood stasis syndrome in patients with coronary artery disease （CAD） complicated with anxiety and depression and explore its underlying mechanisms. MethodsThis study employed a randomized， double-blind， and placebo-controlled clinical trial design. Patients admitted to the hospital were randomly assigned to the observation group and the control group， with 52 patients in each group. Patients in the observation and control groups received Shexiang Baoxinwan and placebo， respectively， both in combination with conventional Western medication. The dose was 45.0 mg， three times daily， for a total duration of eight weeks. The primary outcome was the Seattle Angina Questionnaire （SAQ） scores before and after treatment. Secondary outcomes included changes in traditional Chinese medicine （TCM） syndrome score， the patient health questionnaire-9 （PHQ-9）， generalized anxiety disorder-7 （GAD-7）， inflammatory markers ［interleukin-18 （IL-18）， interleukin-10 （IL-10）， tumor necrosis factor-alpha （TNF-α）， CD40， etc.］， monoamine neurotransmitters ［e.g.， dopamine （DA）］， vascular endothelial function markers ［e.g.， endothelin-1（ET-1）］， adipokines， and ischemia-modified albumin （IMA）. Adverse reactions were also recorded. ResultsA total of 92 patients completed the study， with 44 in the observation group and 48 in the control group. Compared with baseline， both groups showed significant decreases in PHQ-9， GAD-7， and TCM syndrome scores following treatment （P<0.05）， along with a significant increase in SAQ scores （P<0.05）. In the observation group， DA levels were significantly increased （P<0.05）， while levels of IL-18， TNF-α， CD40， ET-1， and IMA were decreased （P<0.05）. In contrast， the control group exhibited significantly increased CD40 levels （P<0.05）. Compared with the control group after treatment， the observation group showed significant improvements in the SAQ dimensions of physical limitation， angina stability， treatment satisfaction， and disease perception， as well as in TCM syndrome score， PHQ-9 score， IL-18， CD40， ET-1， and IMA （P<0.05）. No adverse reactions were observed in either group during treatment. ConclusionShexiang Baoxinwan can improve anxiety and depression， alleviate angina symptoms， and reduce TCM symptoms of Qi stagnation and blood stasis in CAD patients. The mechanism may involve anti-inflammation， improvement of vascular endothelial function， reduction of IMA， and increase of monoamine neurotransmitter levels.

Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
Animals ; Mice ; SARS-CoV-2/genetics* ; COVID-19 ; Vaccines, Synthetic/genetics* ; Genes, Reporter ; Bacteriophages ; Mice, Inbred BALB C

Objective To observe the effects of"Sleep Recipe"on the behavior,brain tissue and central neurotransmitters of insomnia rats.Methods The male rats with SD were randomly divided into control group,model group,sleep formula group,and eszolam group,with 10 rats in each group,and the insomnia model was constructed by intraperitoneal injection of P-chlorphenylalanine(PCPA).After successful modeling,the control group and the model group were given saline gavage,and the medylom group and eszolam group were given drug gavage.The insomnia-like behavior of rats in each group was evaluated by pentobarbital sodium correction experiment and open field experiment,Hematoxlin and eosin(HE)staining observed the pathological changes of rat cerebral cortex,hippocampus,and hypothalamic tissue,and Enzyme-linked mmunosorbent assay(ELISA)was used to determine the expression levels of Gamma-aminobutyric acid(GABA),5-hydroxytryptamine(5-HT).Results The sleep latency of rats in the model group was significantly elongated(P<0.01),while the sleep time was less(P<0.01),and the mental state and fur color were poor,significantly decreased in body weight(P<0.01).Compared with the model group,the sleep latency was significantly shortened(P<0.01),the sleep duration was significantly prolonged(P<0.01),and the body mass was significantly increased(P<0.05,P<0.01);In the open field experiment,the total activity distance of rats in the model group increased,the average speed and central region residence time decreased(P<0.05),the total activity distance of rats in the Sleep Formula group and Eszolam group decreased significantly(P<0.05),the average speed increased and the central region residence time increased(P<0.05).HE results showed that the number of neurons,morphological structure and arrangement of neurons,such as cerebral cortex,hippocampus and hypothalamus in the model group,were damaged to varying degrees,and the sleep formula group and estazolam group were significantly improved.ELISA results showed that the expression of 5-HT and GABA in the cerebral cortex and hypothalamus of rats in the model group was significantly reduced(P<0.01,P<0.05),and the expression of GABA in hippocampal tissues was also significantly reduced(P<0.01).The protein expression levels of cerebral cortical GABA,hypothalamic GABA and 5-HT in the Sleep Formula group were significantly increased(P<0.01).Conclusion Sleep Formula can improve the mental state,restore normal body weight,improve sleep efficiency,and reduce anxiety and tension in insomnia rats.The mechanism may be related to increasing the content of 5-HT and GABA,and inhibiting the spread and conduction of hypothalamic and brainstem pro-awakening nuclei.

Crossmodal information processing in sensory cortices has been reported in sparsely distributed neurons under normal conditions and can undergo experience- or activity-induced plasticity. Given the potential role in brain function as indicated by previous reports, crossmodal connectivity in the sensory cortex needs to be further explored. Using perforated whole-cell recording in anesthetized adult rats, we found that almost all neurons recorded in the primary somatosensory, auditory, and visual cortices exhibited significant membrane-potential responses to crossmodal stimulation, as recorded when brain activity states were pharmacologically down-regulated in light anesthesia. These crossmodal cortical responses were excitatory and subthreshold, and further seemed to be relayed primarily by the sensory thalamus, but not the sensory cortex, of the stimulated modality. Our experiments indicate a sensory cortical presence of widespread excitatory crossmodal inputs, which might play roles in brain functions involving crossmodal information processing or plasticity.
Animals ; Auditory Cortex/physiology* ; Neuronal Plasticity/physiology* ; Neurons ; Rats ; Thalamus ; Visual Cortex/physiology*

BACKGROUND: It has been reported that human adipose-derived mesenchymal stem cells can be induced to differentiate into hepatocyte-like cells with the function of albumin synthesis and urea secretion in vitro.OBJECTIVE: To investigate the potential of adipose-derived mesenchymal stem cells differentiating into hepatocyte-like cells in vitro.METHODS: Adipose-derived mesenchymal stem cells were isolated from the subcutaneous fat of hepatitis B virus infection patients by collagenase digestion and adherent method. Adipose-derived mesenchymal stem cells were induced by three-phase induction method and observed morphologically. The expression levels of alpha-fetoprotein, albumin and cytokeratin 18 were detected by immunohistochemical staining and glycogen synthesis function was detected by glycogen staining method.RESULTS AND CONCLUSION: Most of adipose-derived mesenchymal stem cells induced by three-phase induction method were differentiated into hepatocyte-like cells with polygonal morphology. Immunohistochemistry staining results showed that hepatocyte-like cells expressed alpha-fetoprotein, albumin and cytokeratin 18, and the expression levels of albumin and cytokeratin 18 increased with the culture time. The induced cells had the function of glycogen synthesis and were positive for periodic acid Schiff staining. These results showed that the subcutaneous adipose-derived mesenchymal stem cells could be induced into functional hepatocyte-like cells in hepatitis B virus infection patients.

Objective To analyze the cognitive changes and influencing factors in patients with lacunar cerebral infarction after carotid artery intervention therapy. Methods Sixty lacunar cerebral infarction combined with carotid stenosis patients treated with artery intervention therapy (intervention therapy group) and 68 lacunar cerebral infarction without carotid stenosis patients treated with drug therapy (drug therapy group) were selected. The neuropsychological test was completed at entry and 1, 6, 12 months after entry, and the results were compared with 60 healthy controls (control group). The cognitive changes were observed. The neuropsychological test included mini mental state examination (MMSE), Montreal cognitive assessment scale (MoCA) and cognitive field test. Results There were statistical differences in other scores except the Stroop test C section and Wechsler adult intelligence scale (WAIS-RC) picture arrangement subtest at entry in intervention therapy group and drug therapy group compared with control group (P<0.05). There were no statistical differences in the all scores at entry between drug therapy group and intervention therapy group (P>0.05). In intervention therapy group, the MMSE scores, MoCA total score, Rey-Osterrieth complex figure test (ROCFT), auditory verb learning test (AVLT), and the WAIS-RC picture arrangement subtest, verbal fluency test, WAIS-RC digit span backwards subtest of performing function 12 months after entry were significantly better than those at entry, and there were statistical differences (P<0.05). MMSE score, MoCA total score, long-time delayed recall of ROCFT, the immediate recall, long-time delayed recall and short delayed recall of AVLT, semantic category fluency test of performing function and digit span backwards subtest of WAIS-RC 6 months after entry were significantly better than those at entry:(27.8 ± 2.2) scores vs. (26.4 ± 1.9) scores, (20.7 ± 2.3) scores vs. (19.3 ± 2.0) scores, (12.4 ± 3.2) scores vs. (10.8 ± 2.6) scores, (54.3 ± 10.6) scores vs. (49.9 ± 10.9) scores, (12.4 ± 2.0) scores vs. (11.2 ± 2.8) scores, (12.9 ± 2.0) scores vs. (10.6 ± 2.6) scores, (17.5 ± 4.0) scores vs. (15.4 ± 3.4) scores and (4.0 ± 0.9) scores vs. (3.5 ± 0.9) scores, and there were statistical differences (P<0.05). In drug therapy group, there were no statistical differences in the all scores 1 and 6 months after entry, compared with that at entry (P>0.05);the MMSE score, MoCA total score, ROCFT, the immediate recall, long-time delayed recall and short delayed recall of AVLT, WAIS-RC picture arrangement subtest, verbal fluency test, WAIS-RC digit span backwards subtest of performing function and digit span backwards subtest of WAIS-RC 12 months after entry were significantly better than those at entry, and there were statistical differences (P<0.05). There were no statistical differences in all scores 12 months after entry between intervention therapy group and drug therapy group (P>0.05). In patients intervention therapy group, Logistic regression analysis showed that the MoCA score was related with age, hypertension and low education level (P<0.01 or<0.05), but was not related with smoking, diabetes and interventional treatment (P>0.05). Conclusions Cognitive impairment in patients with lacunar cerebral infarction and carotid stenosis is severe and extensive, but most cognition disorders can improve to normal level 12 months after artery intervention therapy.

Objective Respectively applying the treatment of biventricular pacing and right ventricular septal pacing in atrioventricular block,to compare the heart function influence of two kinds of pacing mode on pacemaker dependent patients, to provide evidence for the physiological pacing mode selection?Methods Enrolled 20 patients from January 2012 to March 2013 who should be placed in pacemakers, their primary disease was the second degree,high or third degree atrioventricular block,giving them three chamber pacemaker ( right atrial + biventricular ) each?Randomly divided into right ventricular septum pacing group ( group A, n=10) and biventricular pacing group( group B,n=10)?Twelve months later,each group crossed into the each other group and continued following?up for 12 months?After 24 months to obtain all the data to do the statistical analysis,including patients'6 min walking distance(6MWD),the Minnesota Heart Failure Quality of life score (MLHFQ),plasma N?terminal pro brain natriuretic peptide precursor(NT?proBNP),left ventricular ejection ejection fraction(LVEF),left ventricular diastolic end diastolic diameter(LVEDD),left ventricular contraction end diastolic diameter(LVESD),left ventricular twelve segmental 14W time standard deviation(Ts?12SD),left ventricular twelve segmental 14W time maximum delay(Ts?dif),the paced QRS qrsd?Results Compared with group B,the 6MWD and LVEF of 12,24 months after treatment of group A were significantly increased( ( 242?58 ±37?56) m vs?(347?42±36?59) m vs?(340?67±24?99) m;(39?97±5?84)% vs?(57?92±10?01)% vs?(60?50±10?06)%;P<0?05),QRSd and NT?proBN were significantly decreased((139?25±10?43) ms vs?(114?25±10?07) ms vs?(110?83±11?08) ms) ms;( 2 857?84±236?48) ng/L vs?( 2 144?26±301?43) ng/L vs? (2 025?91±307?42) ng/L;P<0?05)?Compared with before treatment,at 12 and 24 months after treatment,6MWD in group B was significantly increased(228?17+38?06) m,(329?33+46?28) m,(350?67+35?43) m, LVEF was significantly increased ( ( 40?25+11?24 )% vs? ( 59?50+9?14 )% vs? ( 60?17+10?29)%),QRSd significantly narrowed((142?42+10?66) ms vs?(118?58+994) ms vs?(116?25+10?59) ms) and NT proBNP levels significantly reduced((2 848?25+318?65) ng/L vs?(2 144?26+301?43) ng/L vs?( 2 025?91+30?742) ng/L) were,the difference had statistical significance ( P<0?05)?There was no significant difference on the different time between the groups ( P=0?05)?Comparisons between A and B group at the same treatment time,these indexes of detections were no statistical significance(P>0?05)?Conclusion Compared with the right septal pacing,biventricular pacing is of no significant advantages on the effect of cardiac function for patients with pacemaker dependent.