Lipid nanoparticles(LNPs),composed of ionizable lipids,are currently the most promis-ing non-viral nucleic acid drug delivery vectors in clinical practice,and have great potential in gene ther-apy drug delivery and vaccine delivery.Ionizable lipids,the main components of LNPs,play a decisive role in the endosome escape rate,transfection efficiency,organ targeting and safety of LNPs.Among them,the hydrophilic head group of ionizable lipids contains tertiary amine groups,which can improve the buffering capacity of LNPs and thus change the pKa value,and imidazole can enhance the stability and transfection activity of mRNA-LNP.The ligand contains ester groups,which can induce gene silencing efficiently and improve the degradation rate and safety.When the number of hydrophobic tails is 3-4,with 1-2 unsaturation and 8-18 carbon chain length,LNPs can effectively induce gene silencing.Meanwhile,the presence of branching or asymmetric hydrophobic tails can improve the transfection efficiency of LNPs.Based on the chemical structure of ionizable lipids,this review summarizes the influ-ence of the structure of ionizable lipids on the transfection efficiency and safety of nucleic acid carrying drugs LNPs,and the structure-activity relationship of ionizable lipids so as to provide reference for studies on novel ionizable lipids.

Objective:To investigate therapeutic effect of reinfusion of tumor-infiltrating lymphocyte(TIL)with clustered regularly interspaced short palindromic repeats/CRISPR-associated 9(CRISPR/CAS9)knockout programmed death-1(PD-1)on colon cancer in mice.Methods:Subcutaneous injection of CT26 was used to establish mouse colon cancer model.TIL was extracted from tumor tissue of three model mice,and peripheral blood lymphocytes were extracted.PD-1 gene was knocked out of TIL.Reinfusion experiments were divided into control group(Control),lymphocyte group(Lym),tumor-bearing mouse TIL group(TIL),lentivirus empty empty group(pVSV-G-PX458-NC)and PX458-PD-1-sgRNA1 group(PD-1-sgRNA1),with 10 mice in each group.Tumor tissue quality and tumor inhibition rate were detected in each group.TUNEL was used to detect cell apoptosis in tumor tissues of mice.ELISA was used to detect contents of TNF-α and IFN-γ in tumor tissues of mice.Immunohistochemistry was used to detect expressions of CD4+T and CD8+T cells in tumor tissue.Immunofluorescence was used to detect expressions of proliferating cell nuclear antigen-67(Ki-67)and vascular endothelial growth factor(VEGF).Western blot was used to detect expressions of PD-1 and its ligand PD-L1 in tumor tissues.Results:PD-1-sgRNA1 could significantly inhibit growth of mouse tumor cells in vivo,inhibit expressions of Ki-67 and VEGF in tumor tissues,as well as expressions of PD-1 and PD-L1,elevate apoptosis rate,contents of TNF-α and IFN-γ in tumor tissues,and expressions of CD4+T and CD8+T cells(all P<0.05).Conclusion:Reinfusion of TIL with CRISPR/CAS9 knockout PD-1 can significantly inhibit expressions of Ki-67 and VEGF in colon cancer mice,enhance infiltration of CD4+T and CD8+T cells,induce tumor cell apoptosis and inhibit tumor growth.

Objective:To investigate impacts of usnic acid(UA)on malignant behavior of gastric cancer cells by regulating the chemokine(C-C motif)ligand 2(CCL2)-CCL2 receptor(CCR2)signal axis.Methods:SGC-7901 cells,a well growing human gastric cancer cell line,were treated with different concentrations of UA,which were grouped into low concentration(UA-L)group(62.5 μmol/L UA),medium concentration(UA-M)group(125 μmol/L UA)and high concentration(UA-H)group(250 μmol/L UA);meantime,the cells were transfected with CCL2 overexpression vector(pc DNA3.1 CCL2),empty vector(pc DNA3.1),silenced CCL2(si CCL2)and negative control(si control),and SGC-7901 cells were treated with 250 μmol/L UA,labeled as UA-H+pc DNA3.1 CCL2 group,UA-H+pc DNA3.1 group,UA-H+si control group and UA-H+si CCL2 group,another untreated SGC-7901 cells were taken as the control group.Flow cytometry,MTT and qRT-PCR were applied to detect cell apoptosis,proliferation,and expres-sion levels of CCL2 and CCR2 mRNA;Western blot was applied to detect expression levels of PD-L1,apoptotic protein(Bax),proli-ferative protein(CyclinD1,CCL2,CCR2)and immune escape related protein(B7H1);after co-culturing with CD8+T cells isolated and cultured in vitro,ELISA was applied to detect levels of IL-4,IFN-γ and IL-10 in the supernatant.Gastric cancer cells in each group were co-cultured with activated peripheral blood mononuclear cell(PBMC)1∶1 for 72 hours,and the sensitivity of gastric can-cer cells in each group to T-cell-mediated killing was compared.Results:Compared with control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-L group,UA-M group and UA-H group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels,Bax protein expression were obviously increased(P<0.05);compared with UA-H+pc DNA3.1 group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+pc DNA3.1 CCL2 group were obviously increased,while apoptosis rate,IL-4 and IFN-γ levels,and Bax protein ex-pression were obviously reduced(P<0.05);compared with UA-H+si control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+si CCL2 group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels and Bax protein expression were obviously increased(P<0.05).Conclusion:UA can inhibit gastric cancer cells proliferation,immune escape,and induce apoptosis,which may be related to the inhibition of the CCL2-CCR2 signaling axis.

Diabetes mellitus with ketoacidosis and combined with coma are acute critical complications, which can be complicated with acute abdomen, such as acute pancreatitis, mesenteric thrombosis, small intestine necrosis, etc. There is no report of massive intestine necrosis in the previous literature. We present an overview and aim to improve the diagnosis of acute complications in diabetes mellitus combined with acute abdomen.

This study aimed to evaluate the efficacy of Chinese herbal medicine (CHM) in patients with severe/critical coronavirus disease 2019 (COVID-19). In this retrospective study, data were collected from 662 patients with severe/critical COVID-19 who were admitted to a designated hospital to treat patients with severe COVID-19 in Wuhan before March 20, 2020. All patients were divided into an exposed group (CHM users) and a control group (non-users). After propensity score matching in a 1:1 ratio, 156 CHM users were matched by propensity score to 156 non-users. No significant differences in seven baseline clinical variables were found between the two groups of patients. All-cause mortality was reported in 13 CHM users who died and 36 non-users who died. After multivariate adjustment, the mortality risk of CHM users was reduced by 82.2% (odds ratio 0.178, 95% CI 0.076-0.418; P < 0.001) compared with the non-users. Secondly, age (odds ratio 1.053, 95% CI 1.023-1.084; P < 0.001) and the proportion of severe/critical patients (odds ratio 0.063, 95% CI 0.028-0.143; P < 0.001) were the risk factors of mortality. These results show that the use of CHM may reduce the mortality of patients with severe/critical COVID-19.
Age Factors ; Aged ; COVID-19/therapy* ; China ; Drugs, Chinese Herbal/therapeutic use* ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Odds Ratio ; Propensity Score ; Retrospective Studies ; Survival Rate

Objective To investigate the clinical efficacy of bone grafts containing bone morphogenetic protein 2 (Bone Conduction) treating of nonunion of femoral fracture after intramedullary nailing.Methods The clinical data of 49 patients with nonunion of femoral fracture after intramedullary nailing from August 2008 to August 2016 were analyzed retrospectively.They were divided into experimental group and control group according to different bone grafting methods.The experimental group (n =23) was treated with Bone Conduction combined with allograft bone grafting.The control group (n =26) was treated with autogenous bone grafting.The autogenous bone was taken from the ipsilateral ilium.The two groups were compared operative time,intraoperative blood loss,postoperative wound complications,hospitalization time,postoperative visual analog scale,mean image healing time and the score for distal femoral fractures after the operation.All patients were followed up for 12 to 24 months.Follow-up methods were divided into telephone follow-up and outpatient follow-up.The patients were followed up for outpatients before fracture healing.The patients were followed up by telephone after fracture healing.The healing time of fracture,the flexion and extension activities of the knee joint at 9 months after operation were recorded.SPSS19.0 statistical software was used to analyze the data.Measurement data were expressed as ((x) ±s).The independent sample t test was adopted in measurement data and the counting data was tested by Chi-squared test.Result In the experimental group,operative time was (138.7 ± 34.6) minutes,intraoperative blood loss was (315.4 ± 52.8) ml,postoperative wound complications was 0 case,hospitalization time was (8 ± 3) days,postoperative visual analog scale was 5 scores,but in the control group,operative time was (170.5 ± 32.5) minutes,intraoperative blood loss was (367.2 ± 56.4) ml,postoperative wound complications was 3 cases,hospitalization time was (13 ± 2) days,postoperative visual analog scale was 8 scores.There were significant differences between the two groups,P ＜0.05.All patients who followed up for 12-24 months were recovered,with an average follow-up of 15 months.Mean image healing time in the experimental group and control group was separately(4.9± 2.5) months,(5.2 ± 2.8) months.The excellent and good rates ofthe score for distal femoral fractures 9 months after the operation was separately 95.7％,96.2％,there was no significant difference between the two groups,P ＞ 0.05.Conclusions Bone graft material containing bone morphogenetic protein 2 (Bone Conduction) has the advantages of simple operation,less trauma,less complications,high osteogenic activity and good bone graft effect are equalled with iliac bone graft,the bone graft material containing bone morphogene protein is an effective method to treat nonunion of femoral fracture after intramedullary nail.

Objective To explore the role and possible mechanism of transforming growth factor (TGF)-β autocrine in CDH17 regulating invasion of gastric cancer cells.Methods Construction and transfection of siRNA-CDH17 into MKN-45 gastric cancer cell line to silence the expression of CDH17.Expression of TGF-β and concentrations of TGF-β in supernatants were detected before and after CDH17 silence by immunofluorescence,immunoblotting and ELISA.The autocrine situation of TGF-β was observed.Meanwhile,the activation of TGF-β/Smad3 signaling pathway was also detected by immunoblot.After giving signaling pathway inhibitor,the changes of invasion ability of MKN-45 cells were observed by Transwell invasion experiment.The role of TGF-β autocrine and related signaling pathway activation in CDH17-regulated invasion of gastric cancer cells was evaluated.Results After transfecting siRNA-CDH17 for silencing CDH17 expression in MKN-45 cells,the expression of TGF-β was significantly decreased compared with non-transfection group,its concentration in supernatants was also significantly reduced[(510 ±55)pg/mL vs.(115±20) pg/mL,P<0.01].The immunoblots revealed that phosphorylation level of Smad3 after CDH17 silence was also significantly diminished.However,giving the TGF-β/Smad3 signaling inhibitor SIS3 (10 μmol/L) could also suppress the phosphorylation level of Smad3 when CDH17 was highly expressed,meanwhile silencing CDH17 and inhibiting Smad3 phosphorylation could significantly decrease the invasion of MKN-45 gastric cancer cells (P<0.05).Conclusion CDH17 could participate in the invasion of gastric cancer cells by promoting TGF-β autocrine to activate TGF-β/Smad3 signaling pathway.

Objective To investigate the clinical results and prognosis between proximal and total gastrectomy in proximal advanced gastric cancer.Methods The clinicopathological data of 221 patients with proximal advanced gastric cancer who undcrwent radical gastrectomy were retrospectively analyzed.82 patients and 139 patients underwent proximal and total gastrectomy respectively.The number of dessected lymph nodes,postoperative complications and mortality were compared.Kaplan-Meier survival rate curves and Log-rank test were drawn and compared.Results A total of 1 411 (11-34) and 3 345 (14-35) lymph nodes were harvested from the surgical specimens of 82 and 139 patients with proximal and total gastrectomy respectively,the average number of harvested lymph nodes was 17 ± 11 and 24 ± 10 (t =2.586,P ＜0.05).The overall complications in proximal gastrectomy were higher than total gastrectomy (73.2％ vs.30.2％,x2 =38.291,P ＜0.01).The ratio of functional delayed gastric emptying,anastomotic leakage and stenosis,reflux esophagitis in proximal gastrectomy patients was higher than total gastrectomy.The survival rate in proximal gastrectomy were lower than total gastrectomy in proximal advanced gastric cancer (P ＜0.01).Conclusion Total gastrectomy is recommended for proximal advanced gastric cancer.

ObjectiveTo investigate the clinical features and prognosis of children with rotavirus gastroenteritis and convulsion.MethodsClinical data of children with rotavirus gastroenteritis hospitalized from January 2010 to December 2013 were retrospectively analyzed. Subjects were divided into the seizure group and no seizure group according to the presence of seizure in the course and compared between the two groups.ResultsThere were no signiifcant differences in sex, age, and the average duration of hospitalization between two groups (allP>0.05). The family history, history of seizures, the levels of serum sodium, calcium, lactate, standard bicarbonate concentration (SB), actual bicarbonate concentration (AB), carbon dioxide content (TCO2) and pH were statistically signiifcant between two groups (allP>0.05). During the follow-up period (outpatient telephone follow-up), the recurrence of seizure in two groups was signiifcant different (P<0.05) and only one (0.54%) child in seizure group developed epilepsy.ConclusionThis study showed that rotavirus gastroenteritis with convulsion is a benign clinical course.

Objective To observe the effect of preoperative oral administration of carbohydrate on blood glucose,insulin resistance(IR) and inflammatory reaction after gastrointestinal operation.Methods 48 patients receiving gastrointestinal operation were randomly divided into the study group(n =23)and the control group(n =25).Patients in the study group were orally given 25％ glucose solution 300 ml 3 hours before operation.Before anesthesia induction,gastric contents were aspirated through nasogastric tube to examine its volume and pH.Serum high sensitivity C-reactive protein(hsCRP),fasting blood glucose,insulin level and homeostasis model assessment-insulin resistance(HOMA-IR) were detected before operation and on the first morning after operation between the two groups.Results No anesthesia or operation related complications occurred in either groups.Patients had similar gastric contents volume and the PH value of gastric contents.There was no significant difference in serum hsCRP,fasting blood glucose and HOMA-IR between the two groups before operation.But on the first day,fasting blood glucose,HOMA-IR and hsCRP were significantly lower in the study group than in the control group(6.51 ±1.15 vs 7.49 ±0.57 mmol/L,P =0.038;4.34 ± 1.60 vs 6.09 ±2.81,P =0.043;40.45 ± 27.02 vs 80.02 ± 38.98 mg/L,P =0.03).Conclusion Preoperative oral administration of carbohydrate can obviously lower the postoperative blood glucose level and insulin resistance and alleviate postoperative inflammatory reaction.