OBJECTIVE To investigate the biodistribution of lipid nanoparticles(LNPs)with different surface charges and different particle sizes in mice.METHODS LNPs were prepared using microfluidic technology by incorporating positively charged phospholipids,negatively charged phospholipids,ioniz-able phospholipids,and neutral phospholipids into the formulation to create LNPs with corresponding surface charges.The particle size of the LNPs was controlled by polyethylene glycol(PEG)modifica-tion and measured using dynamic light scattering(DLS)and transmission electron microscopy(TEM),while the surface charge was analyzed using a zeta potential analyzer.The LNPs were labeled with a fluorescent dye,and the mice were intravenously injected with 0.625 μmol·kg-1 of LNPs.At 1,4,12 and 24 h post-injection,the brain,heart,livers,spleen,lungs and kidneys were collected.The fluorescence distribution in different organs was detected using an in vivo imaging system to reflect the distribution of LNPs in various organs.RESULTS Particle size analysis showed that,except the ionizable lipid nanoparticles without PEG modification(LNP-MC3),which had a particle size>200 nm,the particle sizes of positively charged LNPs without PEG modification(LNP-Pos),PEG-modified positively charged LNPs(LNP-Pos-P),PEG-modified neutral LNPs(LNP-Neu-P),PEG-modified ionizable LNPs(LNP-MC3-P),and PEG-modified negatively charged LNPs(LNP-Neg-P)were all<200 nm.Zeta potential analysis revealed that the surface charges of the LNPs were the highest in LNP-Pos,followed by LNP-Pos-P,LNP-MC3-P,LNP-Neu-P,LNP-MC3 and LNP-Neg-P.In vivo imaging results indicated that LNP-Pos-P,LNP-Pos and LNP-MC3-P were primarily distributed in the livers,lungs and kidneys,respectively,while LNP-Neu-P and LNP-Neg-P in the livers,kidneys,and lungs,respectively.The distribution of LNP-MC3-P in the brain,heart,spleen and kidneys peaked at 12 h post-injection,but at 24 h in the livers.The distribution of LNP-Pos-P in the lungs peaked at 1 h post-injection.CONCLUSION LNPs are primarily distributed in the livers.Surface charges influence the second most highly-distributed organs.LNP-Pos-P and LNP-MC3-P are the second most highly-distributed in the lungs,and LNP-Neu-P and LNP-Neg-P in the kidneys.

OBJECTIVE To investigate the biodistribution of lipid nanoparticles(LNPs)with different surface charges and different particle sizes in mice.METHODS LNPs were prepared using microfluidic technology by incorporating positively charged phospholipids,negatively charged phospholipids,ioniz-able phospholipids,and neutral phospholipids into the formulation to create LNPs with corresponding surface charges.The particle size of the LNPs was controlled by polyethylene glycol(PEG)modifica-tion and measured using dynamic light scattering(DLS)and transmission electron microscopy(TEM),while the surface charge was analyzed using a zeta potential analyzer.The LNPs were labeled with a fluorescent dye,and the mice were intravenously injected with 0.625 μmol·kg-1 of LNPs.At 1,4,12 and 24 h post-injection,the brain,heart,livers,spleen,lungs and kidneys were collected.The fluorescence distribution in different organs was detected using an in vivo imaging system to reflect the distribution of LNPs in various organs.RESULTS Particle size analysis showed that,except the ionizable lipid nanoparticles without PEG modification(LNP-MC3),which had a particle size>200 nm,the particle sizes of positively charged LNPs without PEG modification(LNP-Pos),PEG-modified positively charged LNPs(LNP-Pos-P),PEG-modified neutral LNPs(LNP-Neu-P),PEG-modified ionizable LNPs(LNP-MC3-P),and PEG-modified negatively charged LNPs(LNP-Neg-P)were all<200 nm.Zeta potential analysis revealed that the surface charges of the LNPs were the highest in LNP-Pos,followed by LNP-Pos-P,LNP-MC3-P,LNP-Neu-P,LNP-MC3 and LNP-Neg-P.In vivo imaging results indicated that LNP-Pos-P,LNP-Pos and LNP-MC3-P were primarily distributed in the livers,lungs and kidneys,respectively,while LNP-Neu-P and LNP-Neg-P in the livers,kidneys,and lungs,respectively.The distribution of LNP-MC3-P in the brain,heart,spleen and kidneys peaked at 12 h post-injection,but at 24 h in the livers.The distribution of LNP-Pos-P in the lungs peaked at 1 h post-injection.CONCLUSION LNPs are primarily distributed in the livers.Surface charges influence the second most highly-distributed organs.LNP-Pos-P and LNP-MC3-P are the second most highly-distributed in the lungs,and LNP-Neu-P and LNP-Neg-P in the kidneys.

Objective To analyze the distribution bio-relationship of loci related antihypertensive drug efficacy in patients with essential hypertension(EH)in Guiyang region to support the development of clinical guidance for in-dividualized medication of hypertension patients in this region.Methods A total of 406 EH patients who visited the Cardiology Department of Guiyang Hospital from December 2020 to December 2021 were collected as the study sub-jects and ligase sequencing method was applied to detect 7 gene loci of antihypertensive drugs.The distribution of gene loci in EH patients of different genders and from different geographic regions was evaluated.Results The mu-tation frequencies of the seven genes,CYP2D6*10(c.100 C＞T),CYP2C9*3(c.1075 A＞C),ADRB1(c.1165 G＞C),AGTR1(c.1166 A＞C),ACE(I/D),NPPA(T2238C)and CYP3A5*3(A6986G)were 47.29%,5.91%,73.15%,6.65%,34.24%,0.49%,and 69.70%respectively;There was no significant difference in distribution frequency of hypertension drug related gene polymorphisms among different genders.Conclusions The distribution frequency of alleles in 7 antihypertensive drug efficacy related loci is not related to gender of patients.EH patients in Guiyang region are more sensitive to β-blockers and calcium antagonists,but less sensitive to other types of antihypertensive drugs.Therefore,the dose administered should be adjusted appropriately when using β-blockers and calcium antagonists.

Objective To investigate the clinical efficacy of open reduction and internal fixation with K-wires for old Gartland Ⅲ supracondylar fracture in children.Methods A retrospective case series study was conducted on 49 children with old Gartland Ⅲ supracondylar humeral fracture admitted from June 2012 to January 2017.There were 35 boys and 14 girls,with average age of 6.1 years (range,3.4-11.5 years).The duration from initial trauma to operation ranged from 22 days to 60 days (mean,29.8 days).There were five cases of radial nerve injury,one case of median nerve injury,and one case of Volkmann ischemic contracture.All patients were surgically treated with open reduction and internal fixation with K-wires + plaster external fixation.Six patients with nerve injury underwent neurological exploration and release.Nerve partial fracture was seen in one patient during operation,and nerve repair was performed.The fracture healing and complications were observed.At the final follow up,Flynn's criteria was applied to evaluate the elbow joint function.The humerus length and transverse diameter of humeral condyle between the healthy limb and the suffered one were compared to evaluate the humerus growth.Results All patients were followed up for 6-24 months,with an average of 12.5 months.All fractures were healed,with no fracture redisplacement or bone nonunion occurred.The neurological symptoms of six patients with nerve injury disappeared after two months.One patient had superficial skin infection at the incision and recovered after treatment.Six patients (12％) developed cubitus varus deformity after operation.At the final follow-up,the clinical and radiographic outcomes were assessed via Flynn's criteria,and the results were excellent in 37 patients,good in five,fair in one,and poor in six patients,with an excellent and good rate of 86％.There were no significant differences in the humerus length [(189.3 ± 27.8) mm vs.(190.6 ± 28.8) mm] and transverse diameter of humeral condylar [(29.5 ± 3.5) mm vs.(29.7 ± 3.6) mm] (P ＞ 0.05) between the healthy limb and the suffered one,respectively.Conclusion For old Gartland type Ⅲ supracondylar humeral fracture in children,open reduction and internal fixation with K-wires + plaster external fixation can attain satisfactory function recovery of elbow joint and reduce the incidence of cubitus varus deformity,without negative influence on children's humerus growth or development.

Harpagoside (HAR) is believed to be a main compound in Scrophularia ningpoensis which possess a broad of biological activities.Human serum albumin (HSA) has important physiological roles in transportation, distribution and metabolism of many endogenous and exogenous substances in body.It is great significance in pharmacology to investigate the interaction mechanism of HAR and HSA.In this work, the interaction between HAR and HSA was investigated by fluorescence and ultraviolet absorption spectroscopy at different pH (pH=4.0, 7.4, and 9.0) and temperatures (297, 310 and 323 K).The experimental results showed that the HAR could cause the fluorescence quenching of HSA through a static quenching procedure, showing that the HAR regularly quenched the intrinsic fluorescence of HSA, and a decrease in the quenching constant was observed with an increase in temperature.Under different conditions, all the magnitude of binding constants (KA) was larger than 105 L/mol and the number of binding sites (n) in the binary system were approximate to 1.Base on the magnitude of enthalpy and entropy changes, the negative values of ΔG, ΔH and ΔS revealed that the binding of HAR with HSA was spontaneous and exothermic process, and the main interaction forces of the HAR with HAR were van der Waals forces and/or hydrogen bonding interaction.The binding distance (r) between the HAR and HSA was calculated to be about 4.2 nm based on the theory of F(o)rster′s nonradiation energy transfer, which indicated that the energy transfer from HSA to HAR occurred with high possibility.What was more, the synchronous florescence spectroscopy confirmed the conformational changes of HSA during the binding reaction.

OBJECTIVETo test the effect of the c-Met inhibitor cabozantinib in inhibiting infections by Listeria monocytogenes (LM) in mice.

METHODSC57BL/6 mice at 6 weeks of age were subjected to intraperitoneal injection of LM and randomized into 4 groups for treatment with intraperitoneal injection of PBS, intragastric administration of cabozantinib (20 µg/g), intraperitoneal injection of ampicillin (Amp, 20 µg/g), or cabozantinib plus Amp. The survival curves were drawn for each group, and the number of bacteria in the blood and brain tissues was determined; serum IL-10 level and NF-κB p65 level in the cerebrospinal fluid (CSF) were assayed, and Evans Blue (EB) content and pathological changes in brain were examined.

RESULTSCompared with PBS-treated mice, the mice treated with cabozantinib showed a significantly higher survival rate, lower bacterial counts in the blood and brain (P<0.05 or 0.001), lower IL-10 (P<0.05) and NF-κB p65 levels (P<0.01), lower brain EB content (P<0.001), and milder pathological changes in the brain. The blood and brain bacterial counts (P<0.001), IL-10 (P<0.01) and NF-κB p65 levels (P<0.001), and brain EB content (P<0.001) were all significantly lower in mice treated with the combination of drugs than in mice treated with cabozantinib alone.

CONCLUSIONCabozantinib can inhibit LM infection in mice and has important values in developing new anti-intracellular infection drug.

Anilides ; pharmacology ; Animals ; Brain ; microbiology ; pathology ; Injections, Intraperitoneal ; Interleukin-10 ; blood ; Listeria monocytogenes ; drug effects ; Listeriosis ; drug therapy ; Mice ; Mice, Inbred C57BL ; Pyridines ; pharmacology ; Transcription Factor RelA ; cerebrospinal fluid

OBJECTIVETo study the role of outer membrane protein T (OmpT) in the pathogenesis of uropathogenic Escherichia.coli.

METHODSIn cultured human bladder epithelial cell line 5637, we examined the adhesion ability of wild-type (CFT073), ompT gene knockout (COTD), and revertant (pST) strains of E.coli to the cells and the extracellular matrix (ECM). The expressions of the adhesion gene iha and virulence gene iroN were detected by real-time PCR. Murine models of urinary tract infection with the 3 strains were established to evaluate the bacterial burden of the bladder and kidney tissue and bacterial counts in blood. We also detected the expressions of interleukin-6 (IL-6) and IL-8 in the bladder and kidney tissues of the mice.

RESULTThe COTD strain showed a significantly lower cell adhesion rate than CFT073 strain [(4.62∓0.39)% vs (8.81∓1.13)%, P<0.05] with also a lower ECM-adhesion rate [(4.95∓0.59)% vs (8.85∓0.79)%, P<0.05]. The mRNA expressions of iha and iroN in CFT073 strain were 2.1 and 3.8 times that of COTD strain. In the mouse model, the mean bacterial load of CFT073 strain in the bladder tissue was 6.36∓0.06, significantly greater than that of COTD (6.01∓0.07) and revertant (6.29∓0.06) strains (P<0.05); the bacterial load of CFT073 strain in the kidney tissue was also significantly higher than that of COTD strain (6.25∓0.05 vs 5.87∓0.06, P<0.05). In mice infected with the wild-type, knockout, and revertant strains, the detection rates of IL-6, which were identical to those of IL-8, in the inflammatory bladder and kidney tissues were 60%, 12.5%, and 50%, respectively.

CONCLUSIONSOmpT may regulate the expression of the adhesion gene iha and the transferrin gene iroN to affect the adhesion of uropathogenic E.coli to host cells.

Animals ; Bacterial Adhesion ; Bacterial Load ; Bacterial Outer Membrane Proteins ; metabolism ; Cell Line, Tumor ; Escherichia coli Infections ; pathology ; Escherichia coli Proteins ; metabolism ; Gene Knockout Techniques ; Humans ; Inflammation ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Kidney ; microbiology ; Mice ; Peptide Hydrolases ; metabolism ; Receptors, Cell Surface ; metabolism ; Urinary Bladder ; microbiology ; Urinary Tract Infections ; microbiology ; pathology ; Uropathogenic Escherichia coli ; pathogenicity

Objective To study the role of outer membrane protein T (OmpT) in the pathogenesis of uropathogenic Escherichia. coli. Methods In cultured human bladder epithelial cell line 5637, we examined the adhesion ability of wild-type (CFT073), ompT gene knockout (COTD), and revertant (pST) strains of E.coli to the cells and the extracellular matrix (ECM). The expressions of the adhesion gene iha and virulence gene iroN were detected by real-time PCR. Murine models of urinary tract infection with the 3 strains were established to evaluate the bacterial burden of the bladder and kidney tissue and bacterial counts in blood. We also detected the expressions of interleukin-6 (IL-6) and IL-8 in the bladder and kidney tissues of the mice. Results The COTD strain showed a significantly lower cell adhesion rate than CFT073 strain [(4.62±0.39)%vs (8.81±1.13)%, P<0.05] with also a lower ECM-adhesion rate [(4.95 ± 0.59)%vs (8.85 ± 0.79)%, P<0.05]. The mRNA expressions of iha and iroN in CFT073 strain were 2.1 and 3.8 times that of COTD strain. In the mouse model, the mean bacterial load of CFT073 strain in the bladder tissue was 6.36±0.06, significantly greater than that of COTD (6.01±0.07) and revertant (6.29±0.06) strains (P<0.05);the bacterial load of CFT073 strain in the kidney tissue was also significantly higher than that of COTD strain (6.25 ± 0.05 vs 5.87 ± 0.06, P<0.05). In mice infected with the wild-type, knockout, and revertant strains, the detection rates of IL-6, which were identical to those of IL-8, in the inflammatory bladder and kidney tissues were 60%, 12.5%, and 50%, respectively. Conclusions OmpT may regulate the expression of the adhesion gene iha and the transferrin gene iroN to affect the adhesion of uropathogenic E.coli to host cells.

Objective To study the role of outer membrane protein T (OmpT) in the pathogenesis of uropathogenic Escherichia. coli. Methods In cultured human bladder epithelial cell line 5637, we examined the adhesion ability of wild-type (CFT073), ompT gene knockout (COTD), and revertant (pST) strains of E.coli to the cells and the extracellular matrix (ECM). The expressions of the adhesion gene iha and virulence gene iroN were detected by real-time PCR. Murine models of urinary tract infection with the 3 strains were established to evaluate the bacterial burden of the bladder and kidney tissue and bacterial counts in blood. We also detected the expressions of interleukin-6 (IL-6) and IL-8 in the bladder and kidney tissues of the mice. Results The COTD strain showed a significantly lower cell adhesion rate than CFT073 strain [(4.62±0.39)%vs (8.81±1.13)%, P<0.05] with also a lower ECM-adhesion rate [(4.95 ± 0.59)%vs (8.85 ± 0.79)%, P<0.05]. The mRNA expressions of iha and iroN in CFT073 strain were 2.1 and 3.8 times that of COTD strain. In the mouse model, the mean bacterial load of CFT073 strain in the bladder tissue was 6.36±0.06, significantly greater than that of COTD (6.01±0.07) and revertant (6.29±0.06) strains (P<0.05);the bacterial load of CFT073 strain in the kidney tissue was also significantly higher than that of COTD strain (6.25 ± 0.05 vs 5.87 ± 0.06, P<0.05). In mice infected with the wild-type, knockout, and revertant strains, the detection rates of IL-6, which were identical to those of IL-8, in the inflammatory bladder and kidney tissues were 60%, 12.5%, and 50%, respectively. Conclusions OmpT may regulate the expression of the adhesion gene iha and the transferrin gene iroN to affect the adhesion of uropathogenic E.coli to host cells.

OBJECTIVETo investigate MUC2 expression in rat colons induced by probiotics and its effects on the inhibition of E.coli K1 (E44) penetration of the intestinal barrier by probiotics.

METHODSSD rats were subjected to intragastric administration of probiotics, E44, or probiotics +E44 on a daily basis for 7 days, and MUC2 expression in the colons was determined by RT-PCR. MUC2-targeted shRNA (shRNA MUC2) and scrambled shRNA plasmids (shRNA NC) were respectively transfected into Lovo cells, and the efficiency of MUC2 knockdown was determined using qRT-PCR. Competitive exclusion assay was used to evaluate the effects of the probiotics against E44 adhesion and invasion.

RESULTSIntestinal MUC2 mRNA expression was up-regulated in the rats after intragastric administration of probiotics, while E44 administration caused significantly lowered MUC2 expression. MUC2 expression was down-regulated (by 66.7%) by transfection with shRNA MUC2 in Lovo cells as compared with the negative control and mock control cells. The inhibition of E44 adherence and invasion by probiotics was significantly attenuated in transfected Lovo cell culture (in which the relative adhesion and invasion rates of E44 were 56.64% and 66.64%, respectively) as compared with those in the control group.

CONCLUSIONThe up-regulation of MUC2 in rat colons can be one of the mechanisms of the probiotics in antagonizing the translocation of the pathogenic bacteria. Silencing MUC2 expression causes attenuated inhibitory effect of the probiotics on E. coli K1 penetration across human intestinal epithelial cells.

Animals ; Animals, Newborn ; Cell Line, Tumor ; Colon ; drug effects ; metabolism ; microbiology ; Escherichia coli ; pathogenicity ; Escherichia coli Infections ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Humans ; Mucin-2 ; genetics ; Probiotics ; pharmacology ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Rats ; Rats, Sprague-Dawley ; Transfection