Cytopharmaceutical based on macrophages is a breakthrough in the field of targeted drug delivery. However, it remains a challenge to localize and control drug release while retaining macrophage activity and exerting its immunotherapeutic effect. Herein, a localized light-triggered release macrophage cytopharmaceutical (USIP@M) was proposed, which could utilize the tumor targeting and immunotherapy effects of macrophages to reverse the immune suppression of tumor microenvironment (TME). Amphiphilic block copolymers with ultraviolet (UV)-responsive o-nitrobenzyl groups were synthesized and co-loaded with sorafenib (SF), IMD-0354 (IMD), and upconverting nanoparticles (UCNPs), which were then taken up by macrophages, and the targeted delivery of drugs was realized by using the tumor tropism of macrophages. UCNPs converted near-infrared light with strong penetrability and high safety into UV light, which promoted the photoresponsive depolymerization of block copolymers and production of exosomes from USIP@M, accelerated drug efflux and maintained the activity of macrophages. IMD simultaneously polarized carrier macrophages and tumor-associated macrophages to exert the antitumor effect of macrophages, enhance T cell immunity, and alleviate the immunosuppressive state of TME. Synergistically with the chemotherapeutic effect of SF, it could effectively kill tumors. In conclusion, based on the localized light-triggered release strategy, this study constructed a novel macrophage cytopharmaceutical that could localize and control drug release while retaining the activity of macrophages and exerting its immunotherapeutic effect, which could effectively treat solid tumors.

【Objective】 To investigate the effect of exosomes produced by hepatitis B virus (HBV)-infected cells on the phenotype and function of macrophages. 【Methods】 The exosomes secreted by HepAD38 cells, which were capable of producing HBV and HepG2 cells, were collected by ultracentrifugation combined with immunosorbent method.The quality and purity of the extracted exosomes were verified by nanoparticle tracking analysis (NTA), scanning electron microscope and Western blot.The M0 THP-1 macrophages differentiated by PMA were stimulated by HepAD38 derived- or HepG2 derived exosomes.Total RNA and protein samples were collected at different time points after stimulation.Real-time fluorescence quantitative PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect cytokine mRNA and protein expressions, respectively.Meanwhile, neutral red assay was performed to analyze macrophage pinocytosis activity, and a commercial kit was used to measure reactive oxygen species (ROS) in THP-1 macrophages.Human reverse transcription chip detection was performed to obtain the microRNAs profile of the exosomes.And the effect of selected miRNA on macrophages was further confirmed by qRT-PCR. 【Results】 Compared with HepG2-derived exosomes, HepAD38-derived exosomes increased the mRNA and protein expressions of IL-1β, MCP-1 and TNFα significantly.However, no difference of pinocytosis capacity or ROS production was found between the HepAD38-derived exosomes group and HepG2-derived exosomes group.Human reverse transcription chip detection results were verified by KEGG analysis and qRT-PCR, and it was found that miR-6824-3p could also significantly increase the expression levels of IL-1β, MCP-1 and TNFα after high expression. 【Conclusion】 This study found that exosomes produced by HepAD38 cells may stimulate macrophages to produce inflammatory factors such as IL-1β, MCP-1 and TNFα through miR-6824-3p, thereby playing a role in HBV infection.

At present, there have been many clinical trials and systematic reviews/Meta-analysis proving the good clinical efficacy of Shufeng Jiedu Capsules in the treatment of respiratory diseases, while comprehensive discussion is still required. This article overviews and analyzes the systematic reviews/Meta-analysis of Shufeng Jiedu Capsules to provide evidence support for clinical practice. The systematic reviews/Meta-analysis of Shufeng Jiedu Capsules were searched from CBM, Wanfang, CNKI, VIP, PubMed, EMbase and Cochrane Library. The AMSTAR 2 scale and GRADE system were respectively employed for the evaluation of methodological quality and the grading of evidence quality. Finally, 8 systematic reviews/Meta-analysis published during 2018-2021 were included for analysis. The diseases involved include acute exacerbation of chronic obstructive pulmonary disease, community-acquired pneumonia, acute tonsillitis, acute exacerbation of chronic bronchitis and acute upper respiratory tract infection. The number of included RCTs studies ranged from 8 to 25. The results showed that Shufeng Jiedu Capsules combined with western medicine routine had better therapeutic effect than the latter alone in the treatment of the above five diseases. The reported adverse reactions caused by Shufeng Jiedu Capsules were mainly gastrointestinal discomforts such as mild nausea, diarrhoea and vomiting, with low incidence and mild symptoms, which can be relieved by drug withdrawal. The methodological quality of the included studies was extremely low, and the outcome indicators were mainly of low and very low grades. The efficacy and safety of Shufeng Jiedu Capsules in the clinical treatment of diseases still need to be verified based on more high-quality studies. The relevant clinical research and systematic review/Meta-analysis should pay more attention to methodological quality and reporting standards and strengthen the scientificity of research.
Capsules ; Drugs, Chinese Herbal/therapeutic use* ; Systematic Reviews as Topic ; Treatment Outcome

Background: Insulin regulates glucose homeostasis and has important effects on metabolism, cell growth, and differentiation. Depending on the cell type and physiological context, insulin signal has specific pathways and biological outcomes in different tissues and cells. For studying the signal pathway of insulin on glycolipid metabolism in porcine embryonic fibroblast (PEF), we used high-throughput sequencing to monitor gene expression patterns regulated by insulin. Objectives: The goal of our research was to see how insulin affected glucose and lipid metabolism in PEFs. Methods: We cultured the PEFs with the addition of insulin and sampled them at 0, 48, and 72 h for RNA-Seq analysis in triplicate for each time point. Results: At 48 and 72 h, 801 and 1,176 genes were differentially expressed, respectively. Of these, 272 up-regulated genes and 264 down-regulated genes were common to both time points. Gene Ontology analysis was used to annotate the functions of the differentially expressed genes (DEGs), the biological processes related to lipid metabolism and cell cycle were dominant. And the DEGs were significantly enriched in interleukin-17 signaling pathway, phosphatidylinositol-3-kinase-protein kinase B signaling pathway, pyruvate metabolism, and others pathways related to lipid metabolism by Kyoto Encyclopedia of Genes and Genomes enrichment analysis. Conclusions These results elucidate the transcriptomic response to insulin in PEF. The genes and pathways involved in the transcriptome mechanisms provide useful information for further research into the complicated molecular processes of insulin in PEF.

OBJECTIVE: To observe the effect of blistering moxibustion on the expression levels of 5-hydroxytyptamine (5-HT) and its receptors of the colon tissue in the mice with visceral hypersensitivity of irritable bowel syndrome (IBS), so as to explore the effect mechanism of blistering moxibustion in treatment of IBS. METHODS: Forty SPF-grade newborn Kunming mice were randomly divided into a normal group, a model group, an antagonist group and a blistering moxibustion group, 10 mice in each one. Before modeling, the injection with 0.2 mL parachlorophenylalanine (PCPA) was given on the lateral ventricle in the antagonist group. The endorectal glacial acetic acid stimulation combined with tail clipping was used to prepare the model of visceral hypersensitivity of IBS in the model group, the antagonist group and the blistering moxibustion group. After modeling, in the blistering moxibustion group, the intervention with blistering moxibustion was exerted at "Zhongwan" (CV 12), "Tianshu" (ST 25) and "Zusanli" (ST 36), once herbal irritant plaster at each acupoint, for 2 h each time, once a week, consecutively for 3 weeks. Abdominal withdrawal reflex (AWR) score and electromyographic (EMG) amplitude of abdominal muscles were adopted to evaluate the visceral hypersensitivity. HE staining was applied to observe the morphological changes in colon tissue, and immunohistochemistry was to determine the expression levels of 5-HT and its receptors. RESULTS: Compared with the normal group, EMG amplitude of abdominal muscles was increased under 20, 40 mm Hg (1 mm Hg=0.133 kPa) in the model group (P<0.05), AWR scores and EMG amplitude of abdominal muscles under 60, 80 mm Hg were all increased in the model group (P<0.05). In comparison with the model group, EMG amplitude of abdominal muscles was reduced under 20 mm Hg in the blistering moxibustion group (P<0.05), AWR scores were increased under 40 mm Hg in both the blistering moxibustion group and the antagonist group (P<0.05); AWR scores and EMG amplitude of abdominal muscles under 60, 80 mm Hg were all reduced in both the blistering moxibustion group and the antagonist group (P<0.05). Compared with the normal group, in the model group, the mucosa was slightly disturbed, while, the moderate inflammatory cells were visible in the submucosa. In comparison with the model group, the inherent glands of mucosa were regular in shape and a small number of inflammatory cells were visible in both the blistering moxibustion group and the antagonist group. In comparison with the normal group, the average positive staining area percentage (APSAP) of 5-HT and 5-HT₃R of the colon tissue was increased, while, APSAP of 5-HT₄R was reduced in the model group (P<0.05). Compared with the model group, APSAP of 5-HT and 5-HT₃R was reduced in both the blistering moxibustion group and the antagonist group (P<0.05). CONCLUSION Blistering moxibustion can relieve the visceral hypersensitivity of the mice with visceral hypersensitive IBS and the underlying mechanism is related to the regulation of the gut-brain axis mediated by 5-HT signaling pathway.
Animals ; Disease Models, Animal ; Hypersensitivity ; Irritable Bowel Syndrome/therapy* ; Mice ; Moxibustion ; Rats ; Rats, Sprague-Dawley ; Serotonin ; Signal Transduction

Objective To analyze 10 years survival status of urban female patients with corpus uteri cancer and its influencing factors in Liaoning Province. Methods Based on Liaoning cancer register database, 426 patients with corpus uteri cancer in Shenyang, Anshan and Benxi from 2000 to 2002 were randomly selected. They were followed up passively and actively. Life table method and Ederer Ⅱ method were used to calculate the observed survival rate (OSR), the expected survival rate (ESR) and the relative survival rate (RSR). Results We finally included 218 corpus uteri patients. The diagnosis proportions of stage Ⅰ-Ⅳ were 59.2%, 11.5%, 11.0% and 8.7%, respectively. Ten-year RSR and OSR were 59.6% and 67.9%. The diagnosis stage was negatively correlated with 10-year RSR. The 10-year RSR of patients treated with surgery was 71.3%, which was 6.6 times that of non-surgical treatment (10.8%). The 1-year RSR to 10-year RSR ranged from 88.4% to 67.5%. The RSR of each stage was Ⅰ-Ⅱ(95.7%-77.9%) > Ⅲ (71.4%-44.5%) > Ⅳ (58.4%-11.0%). Multivariate Cox model analysis showed that age > 55 years old, late diagnosis stage and non-surgical treatment were the main factors affecting the 10-year survival rate. Conclusion Early diagnosis and surgical treatment can significantly improve the long-term survival rate of patients. Therefore, we should strengthen the early detection and treatment of corpus uteri cancer, standardize and strengthen the screening program.

Aged ; Animals ; Arcobacter/genetics* ; Chickens ; Diarrhea/microbiology* ; Drug Resistance, Bacterial/genetics* ; Genes, Bacterial ; Gram-Negative Bacterial Infections/veterinary* ; Humans ; Male ; Meat ; Microbial Sensitivity Tests ; Phylogeny ; Poultry Diseases/microbiology* ; Virulence ; Virulence Factors/genetics*

Objective:To explore the effects of transcutaneous electric nerve stimulation (TENS) combined with cognitive behavior intervention in the elderly coronary disease patients with hypertension.Methods:From May 2017 to May 2019, we selected 90 elderly coronary disease patients with hypertension in the Second Affiliated Hospital of Harbin Medical University as subjects by convenience sampling. All of patients were divided into control group and observation group with the method of random number table, 45 cases in each group. Control group carried out cognitive behavior intervention, while observation group implemented the TENS combined with cognitive behavior intervention. We compared the angina pectoris symptom control, blood pressure control and the self-perceived burden with the Self-perceived Burden Scale (SPBS) of patients between two groups.Results:After intervention, the effective rate of angina pectoris control of observation group was higher than that of control group with a statistical difference ( P<0.05) ; the diastolic pressure and systolic pressure of observation group were lower than those of control group with statistical differences ( P<0.05) ; the score of SPBS of patients in control group was higher than that in control group also with a statistical difference ( P<0.001) . Conclusions:TENS combined with cognitive behavior intervention for elderly coronary disease patients with hypertension contributes to angina pectoris symptom control and reducing the blood pressure as well as self-perceived burden.

The clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a versatile genome editing tool with high efficiency. A guide sequence of 20 nucleotides (nt) is commonly used in application of CRISPR/Cas9; however, the relationship between the length of the guide sequence and the efficiency of CRISPR/Cas9 in porcine cells is still not clear. To illustrate this issue, guide RNAs of different lengths targeting the EGFP gene were designed. Specifically, guide RNAs of 17 nt or longer were sufficient to direct the Cas9 protein to cleave target DNA sequences, while 15 nt or shorter guide RNAs had loss-of-function. Full-length guide RNAs complemented with mismatches also showed loss-of-function. When the shortened guide RNA and target DNA heteroduplex (gRNA:DNA heteroduplex) was blocked by mismatch, the CRISPR/Cas9 would be interfered with. These results suggested the length of the gRNA:DNA heteroduplex was a key factor for maintaining high efficiency of the CRISPR/Cas9 system rather than weak bonding between shortened guide RNA and Cas9 in porcine cells.
Base Sequence ; Complement System Proteins ; CRISPR-Cas Systems ; DNA ; Genome ; Nucleotides ; RNA, Guide ; Swine

The application of electronic informed consent has the advantages of speeding up the recruitment of subjects, helping the less educated to understand research and recruit groups, facilitating direct communication between subjects and researchers, minimizing human omissions and errors, and effectively improving the efficiency of clinical research. However, there are also some challenges, such as privacy, ethical censorship, hardware costs and so on. On the basis of the above discussion, this paper believed that electronic informed consent, as a new form, would provide a more powerful tool for clinical research in an era full of opportunities and challenges, with research value and potential.