The main clinical manifestation of dyslipidemia is hyperlipidemia， which is an important risk factor leading to the occurrence of cardiovascular and cerebrovascular diseases such as atherosclerosis， coronary heart disease and stroke. In clinical practice， lipid-lowering drugs are mainly used for treatment， but there are issues such as individual differences and genetic effects. Therefore， it is necessary to perform gene detection on patients， so as to guide individualized application of lipid-lowering drugs. This review mainly previews the definition of gene detection and the individualized treatment of lipid-lowering drugs， and introduces the application of gene detection in the individualized treatment of lipid-lowering drugs （statins， fibrates， nicotinic acid and ezetimibe）. Among them， the gene polymorphisms of APOE， SLCO1B1 and CYP450 family play a key role in the efficacy and safety of statins； the gene polymorphisms of APOA/B/C family have a significant impact on the efficacy of fenofibrate； the gene polymorphisms of HCAR2 and DGAT2 have an important impact on the efficacy of niacin； the gene polymorphisms of NPC1L1 have a significant impact on the efficacy of ezetimibe. It is suggested to conduct genotype detection for patients with dyslipidemia to select appropriate treatment strategies， so as to provide individualized medication guidance.

ObjectiveTo obtain HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect by transfecting HSC-T6 cells with CRISPR/Cas9 lentiviral vector, and to provide a good method for further functional research and new strategies for the clinical treatment of liver fibrosis. MethodsThe COX-2 gene-specific sgRNAs (COX-2-sgRNA-1, COX-2-sgRNA-2, COX-2-sgRNA-3) were designed, synthesized, and connected to the GV371 vector, and the recombinant plasmid and the packaging plasmid were transfected into 293T cells to form lentivirus particles; the fluorescence method was used to measure virus titer. The most appropriate amount of the virus was calculated based on MOI. Lenti-Cas9-puro was transfected into HSC-T6 cells, and HSC-T6-Cas9 cells were screened out by puromycin; Lenti-COX-2-sgRNA-EGFP was transfected into HSC-T6-Cas9 cells to obtain HSC-T6-COX-2-/- cells. Cruiser enzyme digestion and Western blot were used to verify gene knockout at the gene and protein levels. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsSequencing verified that the COX-2-sgRNA expression vector was constructed successfully. Recombinant expression plasmids and packaging plasmids were transfected into 293T cells to form lentivirus particles, and the fluorescence method showed a virus titer of ＞1×108. HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect were successfully constructed. The HSC-T6-Cas9 group had significantly higher relative mRNA expression of LV-Cas9-Puro than the CON group (541.93±105.76 vs 1.00±0.02, t=12.995, P＜0.01). Cruiser enzyme digestion and Western blot showed that the CRISPR/Cas9 lentivirus expression vectors played a role in the target, among which COX-2-sgRNA-2 knockout had the most significant effect, and this group had a significant reduction in the protein expression level of COX-2 compared with the CON group and the NC group (both P＜0.05), suggesting that COX-2-sgRNA was active. ConclusionA CRISPR/Cas9 lentivirus vector is successfully constructed for COX-2 target gene, and HSC-T6-COX-2-/- cells with stable COX-2 gene knockout are obtained.

Objective To investigate the virus epidemiological characteristics in patients with acute upper respiratory tract infection, so as to provide a basis for clinical treatment. Methods A total fo 1 306 inpatients or outpatients with acute upper respiratory tract infection in our hospital from January 2017 to January 2020 were enrolled. The respiratory viruses in nasopharyngeal swab samples were detected by fluorescence immunoassay. The detection rate, clinical characteristics, seasonal distribution, and age distribution of each virus were analyzed. Results In 1 306 patients with acute upper respiratory tract infection, 679 cases were positive for virus culture, with a total positive detection rate of 51.99%. Among the single virus infections, 463 cases were positive for FluV, with a positive detection rate of 35.45%. Five different virus infections showed significant difference among 0 ~ years old, 14 ~ years old, 50 ~ years old and 65 ~ years old groups (P<0.05). The positive detection rates of FluV, PIV, RSV, and RV were the highest in the 0 ~ years old group, while ADV detection rate was the highest in the 65 ~years old group. The distribution of the 5 different viruses in spring, autumn and winter was significantly different (P<0.05). Conclusion Acute upper respiratory tract infection is mainly caused by a single virus, and different viruses has significant differences in age and seasonal distribution.

N6-methyladenosine(m

Chloroquine is a quinine derivative which is synthesized by German scholars in 1934. In addition to its anti-malaria, treatment of systemic lupus erythematosus and immunomodulatory effects, chloroquine is also found valuable in broad-spectrum antiviral treatment. Clinical trials have confirmed that chloroquine has a good effect on acquired immunodeficiency syndrome. In 2019, there were many patients infected with novel coronavirus (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2). Preliminary clinical trials showed that chloroquine had obvious curative effect on patients with SARS-CoV-2. We summarize the effects of chloroquine to different viruses, explain its mechanism, and compare the results of its experiments in vitro and in vivo. The antiviral effect of chloroquine in vivo and in vitro are not consistent, which may be related to the model of animal, dosage and distribution of chloroquine in vivo, and the design of clinical research.

ObjectiveTo investigate the effect and significance of cyclooxygenase-2 (COX-2) inhibitors on the expression of the Acsl gene family in the ileum of rats with nonalcoholic fatty liver disease (NAFLD). MethodsA total of 45 Sprague-Dawley rats were randomly divided into normal control group (15 rats given normal diet), NAFLD model group (15 rats given high-fat diet), and nimesulide group (15 rats given high-fat diet and nimesulide). All rats were sacrificed after 12 weeks of feeding, and then blood samples were collected from the inferior vena cava to measure total cholesterol (TC) and triglyceride (TG). HE staining and oil red O staining were performed for the liver to evaluate the degree of hepatic steatosis in each group, and quantitative real-time PCR was used to measure the mRNA expression of the Acsl family genes in the ileum. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal control group, the NAFLD model group had significant increases in serum TC and TG and marked hepatic steatosis (all P＜0.05); compared with the NAFLD model group, the nimesulide group had significant reductions in serum TC and TG and degree of hepatic steatosis (all P＜0.05). Compared with the normal control group, the NAFLD model group had a significant increase in the expression of COX-2 in the ileum (P＜0.05), and compared with the NAFLD model group, the nimesulide group had a significant reduction in the expression of COX-2 in the ileum (P＜005). Compared with the normal control group, the NAFLD model group had significant increases in the mRNA expression of Acsl3 and Acsl5 in the ileum (both P＜0.05), and compared with the NAFLD model group, the nimesulide group had significant reductions in the mRNA expression of Acsl3 and Acsl5 (both P＜0.05). ConclusionThe COX-2 inhibitor nimesulide can regulate the expression of the Acsl gene family in the ileum of rats with NAFLD, suggesting that COX-2 inhibitors may inhibit the progression of NAFLD through the Acsl gene.

OBJECTIVE： To prepare Baicalin-loaded Polyethylene glycol-derivatized phosphatidylethanolamine （BAI@PEG-PE） nanomicelles， and to characterize it and study its cytotoxicity. METHODS： BAI@PEG-PE nanomicelles were prepared by film hydration method and their appearance characteristics were observed. The particle size， polydispersity index， Zeta potential， drug-loading amount and encapsulation efficiency of the nanomicelles were detected. Drug release of BAI raw material and BAI@PEG-PE nanomicelles in pH 7.4 phosphate buffer were compared within 1-84 h. Using coumarin 6 as fluorescent probe， the distribution of PEG-PE nanomicelles in H9c2 cardiomyocytes were observed. H9c2 cardiomyocytes were divided into model group， BAI raw material group and BAI@PEG-PE nanomicelles group. After treated with serum-free DMEM medium containing no or corresponding drugs for 0.5 h， isoproterenol was used to induce cardiomyocyte apoptosis. Nuclear morphology， cell apoptosis rate and protein expression of Bcl-2 and Bax were compared with among 3 groups. RESULTS： Prepared BAI@PEG-PE nanomicelles were uniform globular shape. The particle size was （16.7±0.8） nm， PDI was 0.11±0.01 and Zeta-potential was （－18.4±0.6） mV； drug-loading amount was （7.84±0.65）％， encapsulation efficiency was （85.7±4.9）％ （n＝3）. Accumulative release rate was 76.5％ within 84 h. BAI raw material was released completely within 24 h. PEG-PE nanomicelles could strengthen the intake of coumarin 6 in H9c2 cardiomyocytes， mainly gathering around mitochondria. Compared with model group， the apoptosis morphology of cardiomyocytes were improved significantly in BAI raw material group and BAI@PEG-PE nanomicelles group； apoptosis rate was decreased significantly； protein expression of Bcl-2 was increased significantly； protein expression of Bax was decreased significantly with statistical significance （P＜0.05 or P＜0.01）. Above effects of BAI@PEG-PE nanomicelles group were more significant （P＜0.05 or P＜0.01）. CONCLUSIONS： BAI@PEG-PE nanomicelles are prepared successfully， and show significant sustained-release effect and myocardial targeting， and can prevent cardiomyocyte apoptosis.

Objective] To analyze the medication experience of Zhong Yi-tang for insomnia by using TCM inheritance support system 1.1ver.[Methods] The prescriptions of Zhong Yi-tang for insomnia were collected and input the data into TCM inheritance support system .The frequency and association rules of drugs were calculated by using data mining methods such as apfiofi algorithm,complex system entropy cluster and unsupervised hierarchical cluster,and medication rule of Zhong Yi-tang for epigastric pain was analyzed.[Results] Based on the analysis of 354 prescriptions, the frequency of each herb and association rules among the herbs were computed. Spine date seed, tuber fleeceflower stem, paeony, red sage root, boxthorn and Chinese Date were highly frequently found, 18 pairs of Chinese medicines based on these and 5 core combinations such as ganmai-dazao decoction were mined out . [Conclusion] Zhong Yi-tang usually used ganmai-dazao decoction, spine date seed, tuber fleeceflower stem and red sage root to cure insomnia. The sweet and moistening Chinese medcine were used to strengthen-spleen-stomach, benefiting ying-blood, and nourishing the heart-blood. Data mining method can be used to analyze old TCM doctors’clinical experience.

Liver fibrosis is a common pathological consequence of a variety of chronic stimuli, including viral, autoimmune, drug-induced, cholestatic and metabolic diseases. Fibrosis is driven by a dynamic process involving increased synthesis of matrix components and a failure of physiological mechanisms of matrix turnover. Activation of hepatic stellate cells (HSCs) remains a central event in fibrosis. HSCs are the main source of extracellular matrix (ECM). Transforming growth factor-beta (TGF-Β), which is the fibrogenic master cytokine, can induce the activation of HSCs to produce a large amount of ECM, and is capable of inducing apoptosis of liver cells. RNA interference (RNAi) is a novel gene disruption technology. Studies have shown that small interfering RNA (siRNA) targeting TGF-Β1 may inhibit the activation and proliferation of HSCs, suppress ECM synthesis and block liver fibrosis. TGF-Β1 siRNA-mediated gene silencing therapy provides a new avenue for liver fibrosis. This review summarizes recent progresses in research on HSCs, TGF-Β1 and TGF-Β1 siRNA in liver fibrosis.
Liver Cirrhosis ; therapy ; RNA, Small Interfering ; genetics ; Transforming Growth Factor beta1 ; genetics

BACKGROUNDNeovascular glaucoma is a refractory disease, and difficult to manage. The aim of this study was to evaluate the clinical outcomes of Ahmed glaucoma valve implantation (AGVI) in neovascular glaucoma (NVG) and non-NVG patients.

METHODSThis prospective, non-randomized study included 55 eyes of 55 patients with refractory glaucoma; 27 had NVG (NVG group) and 28 had non-NVG (non-NVG group). All of the patients underwent AGVI. The NVG group was adjunctively injected with intravitreal ranibizumab/bevacizumab (IVR/IVB) before AGVI. Intraocular pressure (IOP) was the primary outcome measure in this study. Surgical success rate, number of antiglaucoma medications used, best corrected visual acuity (BCVA), and postoperative complications were analyzed between the groups.

RESULTSAll of the patients completed the study (follow-up of 12 months). Kaplan-Meier survival curve analysis indicated that the qualified success rates in the NVG and non-NVG groups at 12 months were 70.5% and 92.9%, respectively; this difference was significant (P = 0.036). The complete success rates in the NVG and non-NVG groups at 12 months were 66.7% and 89.3%, respectively (P = 0.049). Compared with preoperative examinations, the postoperative mean IOP and use of medications were significantly lower at all follow-up time points in both groups (all P < 0.05). There were significant differences in BCVA between the two groups at the 12-month follow-up (χ(2) = 9.86, P = 0.020). Cox proportional hazards regression showed NVG as a risk factor for surgical failure (RR = 15.08, P = 0.033). Postoperative complications were similar between the two groups.

CONCLUSIONSAGVI is a safe and effective procedure in refractory glaucoma, but the success rate of surgery was related to the type of refractory glaucoma. The complete and qualified success rates of NVG patient adjunctive anti-vascular endothelial growth factor treatment are still lower than those of non-NVG patients.

Adult ; Aged ; Female ; Glaucoma ; surgery ; Glaucoma Drainage Implants ; Glaucoma, Neovascular ; surgery ; Humans ; Male ; Middle Aged ; Proportional Hazards Models ; Prospective Studies