Objective To evaluate the efficacy of lidocaine solid lipid nanoparticles (L-SLNs) for sciatic nerve blockade in rats.Methods Lidocaine-loaded SLNs were prepared using high pressure homogenization.Ninety SPF male Wistar rats,weighing 220-280 g,were randomized into 6 groups (n =15 each) using a random number table:control group (group C),1％ L-SLN group (group L1-SLN),1％ lidocaine group (group L1),2％ L-SLN group (group L2-SLN),2％ lidocaine group (group L2),and blank SLN group (group SLN).In C,L1-SLN,L1,L2-SLN,L2 and SLN groups,normal saline,1％ lidocaine SLN,1％ lidocaine,2％ lidocaine SLN,2％ lidocaine and blank SLN (200 μl) were injected,respectively,around the sciatic nerve.Before sciatic nerve blockade (baseline) and at 10,20,30,60,120,180,240,300,360,420,480,540 and 600 min after blockade,the paw withdraw latency to a thermal stimulus was measured,and maximum possible effect (MPE) was calculated to reflect the degree of sensory block.Before sciatic nerve blockade and at 10,20,30,60,120 and 150 min after blockade,extensor postural thrust (EPT) of the hind limbs was detected to reflect the degree of motor block.The sciatic nerve at the injection site and the tissues around the site were obtained for observation of the pathological changes at 2 days and 1 and 4 weeks after blockade.Results Compared with the baseline value before blockade and group C,the MPE was significantly increased in at 10-30 min after blockade group L1,at 10-60 min after blockade in group L2,at 10-360 min after blockade in group L1-SLN,and at 10-540 min after blockade in group L2-SLN,and the EPT was decreased at 10-30 min after blockade in group L1,at 10-60 min after blockade in group L2 and group L1-SLN,and at 10-90 min after blockade in group L2-SLN.Compared with group L1,the MPE was significantly decreased at 10 min after blockade,no significant change was found at 20-30 min after blockade,and the MPE was increased at 60-360 min after blockade,and the EPT was increased at 10-30 min after blockade,and no significant change was found at the other time points in group L1-SLN.Compared with group L2,no significant change was found in the MPE at 10-30 min after blockade,the MPE was significantly increased at 60-540 minafter blockade,and the EPT was increased at 10-60 min after blockade,and no significant change was found at the other time points in L2-SLN group.In SLN,L1-SLN and L2-SLN groups,no pathological changes were found in the sciatic nerve at the injection site and the tissues around the site,and only mild inflammatory responses were observed.Conclusion L-SLNs can prolong the duration of block when applied for sciatic nerve blockade in rats and biocompatibility is good.

Acupuncture Points ; Adult ; Female ; Humans ; Male ; Meridians ; Radiculopathy ; diagnosis ; therapy ; Spondylosis ; diagnosis ; therapy

Objective To investigate the toxicity of lidocaine solid lipid nanoparticles (SLNs) in human neurons.Methods Lidocaine-loaded SLNs were prepared using high pressure homogenization.SHSY5Y cells were cultured in vitro and inoculated on 96-well plates (100 μl/well) at a density of 5× 105 cells/ml.SH-SY5Y cells were randomized into 10 groups (n =30 each) using a random number table:control group (group C), different concentrations of lidocaine groups (L1-4 groups), different concentrations of lidocaine SLN groups (L-SLN1-4 groups), and blank SLN group (group SLN).The cells were cultured routinely in group C.The cells were incubated with the culture medium containing lidocaine with the final concentrations of 1.000％, 0.500％, 0.250％ and 0.125％ in L1-4 groups, respectively.In LSLN1-4 groups, the cells were incubated with the culture medium containing lidocaine SLNs with the final concentrations of 1.000％, 0.500％, 0.250％ and 0.125％ in L1-4 groups, respectively.Before incubation (at the corresponding time points in group C), and at 1, 12 and 24 h of culture or incubation (T0-3) , 6 wells in each group were selected for measurement of the cell survival rate (using methyl thiazolyl tetrazolium assay).The cell morphology was examined with optical microscope at T3.Results Compared with that at T0, the cell survival rate was significantly decreased at each time point in L1-4 and L-SLN1,2 groups, at T2,3 in L-SLN3 group, and at T3 in L-SLN4 group (P＜0.05) , and no significant change was found in SLN and C groups (P＞0.05).The cell survival rate was significantly lower at T2,3 in L1-4 and L-SLN1-3 groups, and at T3 in group L-SLN4 than that at T1, and at T3 in L1-4 and L-SLN1-4 groups than that at T2 (P＜0.05).Compared with group C, the cell survival rate was significantly decreased at each time point in L1-4 and L-SLN1,2 groups, at T2,3 in group L-SLN3, and at T3 in group L-SLN4 (P＜0.05) , and no significant change was found in group SLN (P＞0.05).Compared with group L-SLN at the corresponding concentration, the cell survival rate was significantly decreased at each time point in group L1-4 (P＜0.05).Conclusion Lidocaine SLNs have toxic effect on human neurons, but the effect is weaker than that caused by Iidocaine solution.

Objective:To investigate effects of viola on macrophage TOLL-like receptor expression ,and tentatively explore the partial mechanism of viola intervention on macrophage function.Methods: Using viola water decoction lavage intervention in clean grade SD rats of conventional preparation containing serum ,In a certain concentration coculture with mouse peritoneal macrophages in 96-well plates.After 24 hours,cell phagocytosis activity was determined by neutral red uptake assay ,changes in expression levels of TLR-1,TLR-2,TLR-3,TLR-4,TLR-5 mRNAs were determined by RT-PCR.Results:Compared with the same concentration of normal serum group :(1 ) viola-containing serum group macrophage activity was significantly increased ( P<0.01 );( 2 ) Within certain concentration,viola containing serum group with increased or decreased expression levels of TLR -1,TLR-2,TLR-3 mRNA(P<0.05 or P<0.01 ) ,the expression level of TLR-4 was not significantly altered in each group ( P>0.05 ) ,and the expression level of TLR-5 was significantly increased in each group ( P<0.01 ).Conclusion:Viola-containing serum has obvious promoting effect on macrophage phag-ocytosis,the mechanism of which may be related to that this drug regulates and controls part of macrophage TOLL -like receptor expres-sion.

OBJECTIVETo investigate biological transport of minocycline by human periodontal ligament fibroblasts (HPDLF). To verify the hypothesis of delivering medicine to periodontium and the whole body through the root canal.

METHODSHPDLF and MC3T3-E1 cells were incubated in minocycline solutions. The intracellular antibiotics contents were measured by high performance liquid chromatography (HPLC) and the cell total protein was measured by Bradford protein assay.

RESULTSHPLC was an accurate, sensitive method for measurement of the intracellular minocycline. The incubation time and cell property had significant effect on the intracellular minocycline contents (P< 0.01). The intracellular contents increased with extracellular concentration.

CONCLUSIONMinocycline can be transported by HPDLF. The transport is concentration-dependent, time-dependent and cell specific.

Anti-Bacterial Agents ; Biological Transport ; Cells, Cultured ; Fibroblasts ; Humans ; Minocycline ; Periodontal Ligament

Objective To investigate biological transport of tetracycline hydrochloride by human periodontal ligament fibmblasts(HPDLF) for verifying the hypothesis of delivering medicine to the periodonfium and whole bodv through the root canal.Methods HPDLF and MC3,13-E1 cells were ineubated in antibiotics solutions. The intracellulaF antibiotics contents were measured by high performance liquid chromatography(HPLC) and the cell total protein was measured by bradford protein assay.Results The intracellular contents increased with ineubation time. The extracellular medicine concentration had effect on the intracellular contents. Conclusions Tetracycline hydrochloride can be transported into HPDLF with incubation and this transport is time-dependent and concentration-dependent.

OBJECTIVETo generate recombinant human tissue factor pathway inhibitor (TFPI) in Pichia pastoris.

METHODSTo improve the expression of TFPI, a silent mutation was generated at the specific site of TFPI cDNA. Both wild-type TFPI cDNA and mutated TFPI cDNA were cloned into the expression vector pPic9. The constructed plasmids were subsequently transformed into Pichia pastoris cells GS115 and KM71, and the transformants were confirmed by polymerase chain reaction and DNA sequencing. The expression of recombinant protein was induced by addition of 0.5% methanol in the culture medium. The cell culture medium after induction was concentrated through ultra filtration. The recombinant protein was further purified by a three-step process (Heparin-sepharose CL-6B affinity chromatography, DEAE-Sepharose Fast Flow affinity chromatography, and Sephadex G75-gel filtration). The amount of the recombinant protein was quantified with gel imaging system. The activity of the recombinant protein was analyzed by the chromogenic substrate assay.

RESULTSThe amount of TFPI expressed in the mutated clone (1 mg/L) was much higher than that in the wild type clone (0.1 mg/L). The TFPI activity in the recombinant GS115 cells could be detected 12 hours after induction and reached the peak at 36 hours, while the TFPI activity in the recombinant KM71 cells started to show up at 24 hours after induction and reached the peak at 72 hours. The expression of recombinant protein in the silent mutant was significantly higher than those of wild type clone in both GS115 and KM71 host cells. The relative molecular mass of recombinant TFPI was approximately 42 000.

CONCLUSIONIntroduction of the silent mutation at the specific site of TFPI cDNA can increase the recombinant protein expression in Pichia pastoris, which is much higher than that in insect cells or saccharomyces cerevisiae.

Humans ; Lipoproteins ; biosynthesis ; genetics ; Mutation ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo explore the association of gene polymorphism of organophosphate insecticides (OPs) metabolic enzymes with intermediate myasthenia syndrome (IMS) following acute OPs poisoning.

METHODSThirty six of 147 acute OPs poisoning patients developed IMS one to four days after poisoning. Peripheral blood samples were collected from all the patients and whole blood cholinesterase (ChE) activity was determined by DTNB spectrometry. The genetic polymorphism of CYP2E1 (1091C-->T) and GSTP1 (313A-->G) were analyzed by polymerase chain reaction (PCR)-restrict fragment length polymorphism, CYP1A1 (4889A-->G), GSTM1 and GSTT1 by allele-specific PCR, and PON1 at 55 codon (55L-->M) by PCR-single strand conformation polymorphism.

RESULTSThe whole blood ChE activity in IMS patients was not significantly different from non-IMS patients at admission (38.22 +/- 17.56)% and (42.49 +/- 16.23)%, respectively, P > 0.05, but recovered much slower in IMS patients than that in non-IMS patients. The frequencies of heterozygote and variant homozygote of PON1 at 55 codon, GSTM1 null, and both GSTM1 and GSTT1 null were higher in IMS patients than those in non-IMS patients (P < 0.05), with odds ratios and their 95% confident intervals of 2.48 (1.06 - 5.78), 11.23 (2.95- 42.76), 2.53 (1.14 - 5.61) and 2.68 (1.20 - 5.97), respectively.

CONCLUSIONSPatients of OPs and its mixture poisoning with genotype of variant allele at 55 codon of PON1, GSTM1 null and both GSTM1 and GSTT1 null probably had higher risk for IMS.

Adult ; Cholinesterases ; metabolism ; Cytochrome P-450 CYP2E1 ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Glutathione Transferase ; genetics ; Homozygote ; Humans ; Insecticides ; poisoning ; Male ; Middle Aged ; Myasthenia Gravis ; chemically induced ; genetics ; Organophosphorus Compounds ; Point Mutation ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Syndrome

OBJECTIVEThe purposes of our study were to investigate the association between maternal urinary phthalate metabolites and the levels of inhibin B (INHB) and insulin-like factor 3 (INSL3) in the cord blood in a Chinese pregnant population.

METHODSMaternal urine samples in the third trimester of pregnancy of 69 participants were collected and stored, and the samples of cord blood (10 ml) were collected at delivery between June 2011 and September 2012 in a comprehensive hospital of gynecology and obstetrics in Tianjin, China.Four phthalate metabolites, monomethyl phthalate (MMP), monoethyl phthalate (MEP), monobutyl phthalate (MBP), and mono-2-ethylhexyl phthalate (MEHP) were measured in the urine samples using liquid chromatography-tandem mass spectrometry. The levels of INHB, INSL3 in the cord blood were tested by ELISA. Associations of phthalate exposure with INHB and INSL3 levels were determined by spearman correlation and multiple regression model analysis.

RESULTSThe median concentrations of observed metabolites in descending order were 49.74 µg/L for MMP, 24.96 µg/L for MEHP, 19.52 µg/L for MEP and 17.73 µg/L for MBP. The median concentrations of INHB and INSL3 were 89.09 and 106.21 ng/L.Significant negative associations between INHB and MMP(β' = -0.252), MEP(β' = -0.363) or the sum value (∑PAEs) (β' = -0.346) were found by the multiple regression model analysis. For INSL3, only the sum value (β' = -0.313) was inversely significantly associated with the levels of INSL3 in the cord blood.

CONCLUSIONSMaternal urinary phthalate metabolites were associated with INHB and INSL3 in the cord blood in a Chinese population.

Adult ; Diethylhexyl Phthalate ; analogs & derivatives ; urine ; Female ; Fetal Blood ; chemistry ; Humans ; Infant, Newborn ; Inhibin-beta Subunits ; blood ; Insulin ; blood ; Male ; Maternal Exposure ; Phthalic Acids ; urine ; Pregnancy ; Proteins ; Testicular Hormones ; blood ; Young Adult

Objective:To investigate the influencing factors of participation in free health check-up among community residents.Methods:From 2012 to 2022 Xinzhuang Community Health Service Center of Shanghai Minhang district provided 5 free health check-up for local residents, once every 2 years. Among 5 904 eligible community residents with a mean age of (66.01±5.87) years, 682 (11.55 %), 912 (15.45 %), 842 (14.26 %), 934 (15.82 %), 1 061 (17.97 %) and 1 473 (24.95 %) participated in 5, 4, 3, 2, 1 and 0 health check-ups during 10 years, respectively. The influencing factors of participant frequency were analyzed with multivariate logistic regression model.Results:Multivariate logistic regression analysis showed that age 65 years and above ( OR=0.685, 95% CI: 0.625-0.751, P<0.001), exercising once a week or more ( OR=1.142, 95% CI: 1.031-1.266, P=0.011), and underweight ( OR=0.665, 95% CI:0.496-0.891, P=0.006) were independent factors influencing the participant in free health check-up among community residents. Conclusion:Community residents with older age or underweight are less likely to participate free health check-up, while those with frequent exercise like to participate.