Objective To investigate the effects of ropivavaine on gamma -aminobutyric acid(GABA)-activated membrane cur-rents in isolated dorsal root ganglion(DRG) neurons of the rats with ischiadic nerve chronic constriction injury (CCI) and to discuss the possible analgesia mechanism of ropivacaine .Methods The whole-cell patch clamp technique was used to record and compare the changes of GABA receptor activation currents of acute isolated DRG neurons after 30 s of ropivacaine preperfusion in the oper-ating side and the operative opposite side of the CCI model rats and the sham-operation group .Results (1)Compared with the oper-ative opposite side ,the sham-operation group and the control group ,the thermal withdrawal latency in the operative side group of the CCI model rats was notablely shortened(P<0 .05);(2)the amplitude of GABA-activated currents with different concentration GABA(0 .1-1 000μmol/L) in the operative opposite side group of the CCI operation was significantly greater than that of the op-erative side group and the sham-opeartion group ;(3)DRG neurons after ropivacaine preperfusion (0 .1-1 000μmol /L) showed va-rying degrees of enhancement effect on the 100 μmol/L GABA-activated currents ,the enhancement amplitude in the CCI operative opposite side group was significantly greater than that in the operative side group and the sham-operation group ;(4)The dose-re-sponse curve of DRG neurons GABA (0 .1-1 000μmol/L) activated current in the operative side group of the CCI rats after ropiva-caine pre-perfusion (100 μmol/L) was shifted to the left ,the difference between two EC50 had no statistical significance (P>0 .05) .Conclusion Ropivacaine has the enhancement effect on GABA activated currents in the DRG neurons of the CCI model rats , which could be one of reasons for ropivacaine producing the anesthetic and analgesic effect .

Animals ; Antioxidants ; physiology ; Dehydroepiandrosterone ; physiology ; Lipid Metabolism ; physiology ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effects of midazolam on GABAA receptor-activated currents in isolated dorsal root ganglion (DRG) neurons in rats.Methods Sprague-Dawley rats of both sexes,weighing 200-250 g,aged 4 weeks,were used in the study.The DRG neurons were isolated and GABAA receptor-activated currents were recorded using the whole-cell patch-clamp technique.GABAA receptor-activated currents were recorded after administration of the mixture of midazolam 3.00 μmol/L (final concentration)and the different final concentrations (0.03,0.10,1.00,10.00,100.00 and 1000.00 μmol/L) of GABA,after different concentrations of midazolam (0.03,0.10,1.00,3.00,10.00 and 100.00 μmol/L) was given,after administration of the mixture of different final concentrations(0.03,0.10,1.00,3.00,10.00 and 100.00 μmol/L) of midazolam and GABA 100.00 μmol/L (final concentration),and after administration of the mixture of midazolam 1.00μmol/L (final concentration) and GABA 100.00 μmol/L (final concentration)at the preset time points of perfusion with different concentrations of midazolam (0,20,40,60 and 120 s of perfusion).The enhancement rate of the currents was calculated.Results No change in the membrane currents was found after midazolam was perfused in the neurons sensitive to GABA.GABAA receptor-activated currents were enhanced after administration of the mixture of different concentrations of GABA and midazolam.GABAA receptor-activated currents were enhanced after different concentrations of midazolam were given compared with that before administration,and the enhancement rate of the GABAA receptoractivated currents was gradually increased with the increase in the concentration of midazolam and reached the peak at the concentration of 3.00 μmol/L.The enhancement rate of the GABAA receptor-activated currents was gradually increased with the prolongation of perfusion time and peaked at 40 s of perfusion.Conclusion Midazolam can enhance the GABAA receptor-activated currents in rat dorsal root ganglion neurons,indicating that midazolam increases the role of GABA through increasing the activity of GABAA receptors and has analgesic effect at the spinal cord level.

ObjectiveTo investigate the effects of over expression of miR-34a on cellular proliferation and migration in bladder cancer cell line J82 by targeting Notchl.MethodsmiR-34a was predicted as a putative gene which can target Notchl through bioinformatics analysis,qRT-PCR and Western blot were performed to measure the expression levels of Notchl and miR-34a in invasive transitional cell carcinoma of bladder (TCCB) tissues and J82 cells transfected with miR-34a.Luciferase assay was employed to determine if miR-34a could target Notchl through binding to the 3'-untranslated region (3'UTR) of Notchl mRNA.J82 cells were transfected with pcDNA3.0-miR-34a or pcDNA3.0 control plasmid.MTS colorimetry was used to evaluate the effect of miR-34a on cell proliferation.The effect of miR-34a on cell migration was assessed by transwell migration assay.ResultsThe expression level of miR-34 in invasive TCCB tissues was lower than in adjacent bladder tissues (0.016(0.018) vs 0.042 (0.059),N =16; P =0.0006).On the contrary,the average levels of Notchl mRNA and protein were higher in tumors than in adjacent bladder tissues (2.765(2.156) vs 2.312(1.365),N =16; P =0.0025 and 0.857 ±0.197 vs 0.648 ±0.171 ;P ＜0.0001 ).After the transfection of miR-34a,the expressive level of miR-34a in J82 was highly induced ( (2.408 ±0.789) × 10-4 vs(0.153 ±0.029) × 10-4; P =0.0026).However,the expressive levels of Notchl mRNA and protein were obviously decreased (3.001 ± 0.106 vs 4.998 ± 1.053 ; P =0.0308 and 0.747 ± 0.050 vs 0.988 ± 0.102 ; P =0.0215 ).The results of luciferase assay showed that firefly activity was highly dimished (0.422 ± 0.028 vs 2.392 ± 0.148 ; P ＜ 0.0001 ).Cellular proliferation was inhibited after the transfection of miR-34a in J82 (P ＜ 0.0001 ).Moreover,number of migration cells of J82 was significantly reduced after the ectopic expression of miR-34a ( 179.3 ± 21.02 vs 269.7 ± 23.71 ; P =0.0078 ).ConclusionsmiR-34a inhibits the cellular proliferation and migration of bladder cancer cell line J82 via binding to the 3UTR of Notchl mRNA.

OBJECTIVETo investigate the effects of ropivacaine on Gamma-aminobutyric acid(GABA)-activated currents in dorsal root ganglion (DRG) neurons in rats and discuss the analgesia mechanism of ropivacaine.

METHODSBy means of using whole-cell patch-clamp technique, to investigate the modulatory effects of ropivacaine on GABA-activated currents (I(GABA)) in acutely isolated dorsal root ganglion neurons.

RESULTS(1) In 48 out of 73DRG cells (65.7%, 48/73), to perfusion ropivacaine bromide (0.1 - 1 000 micromol/L) were sensitive. Which produce in 0 to 380 pA current. (2) The majority of the neurons examined (74.5%, 73/98) were sensitive to GABA. Concentration of 1 - 1 000 micromol/L GABA could activate a concentration-dependent inward current, which manifested obvious desensitization, and the inward currents could be blocked byGABA-receptor selective antagonist of bicuculline (100 micromol/L). (3) After the neurons were treated with ropivacaine (0.1 - 1000 micromol/L) prior to the application of GABA (100 micromol/L) 30 s, GABA currents were obviously increased. Ropivacaine could make dose-response curve of the GABA up, EC50 is 23.46 micromol/L. Ropivacaine shifted the GABA dose-response curve upward and increased the maximum response to the contrast about 153%.

CONCLUSIONThe enhancement of ropivacaine to DRG neurons activation of GABA current, can lead to enhancement of pre-synaptic inhibition at the spinal cord level. This may be one of the reasons for the anesthetic effect and analgesia for ropivacaine in epidural anesthesia.

Amides ; pharmacology ; Animals ; Ganglia, Spinal ; cytology ; physiology ; Membrane Potentials ; drug effects ; Neurons ; cytology ; drug effects ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Receptors, GABA-A ; physiology

Objective:To explore the application and key points of microchannel approaches in resec-tion of cervical intraspinal tumors.Methods:A retrospective analysis was performed on 51 cases of cervi-cal spinal canal tumors from February 2017 to March 2020.Among them,5 cases were located epidural space,6 cases were located epidural and subdural space,and 40 cases were located under the subdural extramedullary space(6 cases were located on the ventral side of the spinal cord).The maximum diameter ranged from 0.5 to 3.0 cm.The clinical manifestations included neck,shoulder or upper limb pain 43 cases,sensory disturbance(numbness)in 22 cases,and limb weakness in 8 cases.The micro-channel keyhole technique was used to expose the tumor,and the tumor was resected microscopically.Results:In this study,35 patients underwent hemilaminectomy,12 patients underwent interlaminar fenestration,2 patients underwent medial 1/4 facetectomy on the basis of hemilaminectomy or interlaminar fenestration.Two tumors were resected through anatomy space(no bone was resected).The degree of tumor resection included total resection in 50 cases and subtotal resection in 1 case.The type of the tumor included 36 schwannomas,12 meningiomas,2 enterogenic cysts and 1 dermoid cyst.There was no infec-tion and cerebrospinal fluid leakage postoperatively.Limb numbness occurred in 7 patients.The average follow-up time was 15 months(3 to 36 months).No deformity such as cervical instability or kyphosis was found.The tumor had no recurrence.Conclusion:The cervical spinal canal is relatively wide,cervical tumors with no more than three segments can be fully exposed by means of microchannel technology.Besides intramedullary or malignant tumors,they can be microsurgically removed.Preservation of the skeletal muscle structure of cervical spine is beneficial to recover the anatomy and function of cervical spine.The electrophysiological monitoring helps to avoid spinal cord or nerve root injury.

In the present study, an ultra performance liquid chromatography coupled with time-of-fight mass spectrometry (UPLC-TOF/MS) based chemical profiling approach was used to evaluate chemical constitution between co-decoction and mixed decoction of ginseng and Radix Aconiti Praeparata. Two different kinds of decoctions, namely co-decoction of ginseng and Radix Aconiti Praeparata: water extract of mixed two herbs, and mixed decoction of ginseng and Radix Aconiti Praeparata: mixed water extract of each individual herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-TOF/MS analysis. The datasets of t(R) m/z pairs, ion intensities and sample codes were processed with supervised partial least squared discriminant analysis (OPLS-DA) to holistically compare the difference between these two decoction samples. Significant difference between the two decoction samples was showed in the results of positive ion mode. The contents of hypaconitine and deoxyaconitine decreased, while that of benzoylmesaconine, benzoylhypaconine and dehydrated benzoylmesaconine increased in the samples of co-decoction of ginseng and Radix Aconiti Praeparata. The content of diester-diterpenoid alkaloids decreased, while that of monoester-diterpenoid alkaloids increased, which is probably the basis of toxicity-attenuated action when combined ginseng with Radix Aconiti Praeparata.
Aconitine ; analogs & derivatives ; analysis ; Aconitum ; chemistry ; Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

Objective To explore the feasibility and safety of laparoscopic pyeloplasty in the treatment of neonatal renal pelvis ureteraljunction obstruction(PUJO),and to analyze its preliminary experience.Methods From Jun.2009 to Apr.2012,11 neonates(9 boys and 2 girls) were recruited in this study.They were all detected by prenatal ultrasound and renal pelvis anteroposterior diameter was more than 3 cm.One week after birth,emission computed tomography(ECT) showed that split function of hydronephrotic kidneys were lower than 40％.All patients underwent laparoscopic dismembered pyeloplasty.Results All successful underwent laparoscopic dismembered pyeloplasty,no conversion to open surgery or additional Trocar,no intraoperative complications.The mean time of operation was 80 min,and blood loss was less than 10 mL,the mean postoperative hospital stay was 9 days.All patients were followed up for 6 to 36 months with ultrasound and ECT.The thickness of renal parenchyma increased,and 8 kidneys turned almost normal,the other's renal pelvis anteroposterior diameter was about 1.5 cm,renal pelvis anteroposterior was significantly reduced,and the scar was not obvious.Conclusions Laparoscopic ureteroplasty in the treatment of neonatal hydronephrosis is safe and feasible,and it is worthy of application in a large scale.Qperators need mastering laparoscopic suture technology,and then apply the technique from older children to neonates gradually.

Objective To investigate the level and relevant factors of pyrethroid pesticide exposure among pregnant women during the first trimester in rural areas of Yunnan province,and to provide basic data for further study and provide a basis for the formulation of intervention measures to reduce exposure.Methods According to the birth cohort of rural Yunnan built in 2014, pregnant women in the first trimester were recruited in two townships of an agricultural county who were confirmed pregnant and continued pregnancy in antenatal clinics. The concentration of the metabolites of the pesticides in the urine of women was determined by super high performance liquid chromatography and the contact status of pyrethroid pesticides was analyzed using a questionnaire survey. Results A total of 419 women in the first trimester were surveyed and 94.51% of women detected at least one of the pyrethroid metabolites, among which the detection rates of 3－Phenoxybenzoic acid (3－PBA), cis－/trans－3－ (2, 2－Dichloroethenyl)－2, 2－dimethyl－cyclopropanecarboxylate (cis－/trans－DCCA) and 4－fluoro－3－phenoxybenzoic acid (4－F－3－PBA) were 79.47%, 62.05% and 24.58%,and the median concentrations were 3.53 ng/mL, 14.80 ng/m L and less than the minimum detection limit 0.4 ng/m L.Only 12 people (2.95%) reported that they had exposure to pesticides.Self－reported exposure rate was not consistent with the actual exposure rate.Multivariate linear regression analysis showed that the season was a factor affecting the level of three metabolites in women in the first trimester. Higher lever was found in women in spring (t=2.147,P=0.032), summer (t=2.144,P=0.035) and winter (t=2.453,P=0.015) compared to those in autumn.Conclusion Pyrethroid pesticides were widely exposed in the pregnant women in rural areas of Yunnan province. Some women have higher exposure level, which is related to the season.

OBJECTIVETo understand the epidemic characteristics and viral antibody level among healthy people of Japanese B encephalitis (JE) in Henan province in 2010.

METHODSA total of 433 selected JE cases in Henan province in 2010 were analyzed by descriptive epidemiological method. Xinyang and Luoyang were selected as survey sites in 2010.12 administrative villages were randomly selected from both cities. As the investigation objects, 519 healthy people from the two cities were randomly selected by eight age groups:less than one year old, 1 - 2 years old, 3 - 4 years old, 5 - 6 years old, 7 - 14 years old, 15 - 19 years old, 20 - 59 years old, and above 60 years old. A total of 1008 samples of blood specimens were collected both in May and November, and JE viral antibody was detected by micro-cytopathic effect neutralization test.

RESULTSThe incidence rate of JE was 0.46/100 000 (433/94 130 434) in Henan province in 2010.97.69% (423/433) of the patients were found between July and September, and 81.06% (351/433) were distributed in Nanyang, Xinyang, Luoyang, Zhumadian and Zhoukou city. Children aged 0 to 14 years were the primarily affected group (82.22%, 356/433), the people above 15 years old accounted for 17.78% (77/433) of whole cases in Henan province, but the same group accounted for 65.79% (50/76) of whole cases in Luoyang city, which obviously higher than the percentage in Henan province (χ(2) = 79.57, P < 0.05). Most patients were scattered children in Henan province, accounting for 58.89% (255/433). In Luoyang city, most patients were peasants, accounting for 44.74% (34/76). The antibody positive rate of JE among health people above 15 years old in Luoyang city was 48.94% (46/94), which was lower than it in Xinyang city at 97.78% (88/90). The difference showed statistical significance (χ(2) = 55.42, P < 0.05). The antibody positive rate among healthy people under vaccination was 50.41% (61/121), which was obviously higher than that among people without vaccination, at 16.67% (6/36) in Luoyang city. The difference showed statistical significance (χ(2) = 12.92, P < 0.05). The antibody positive rate among healthy people under vaccination was 67.11% (51/76) in Xinyang city, which was obviously higher than that among people without vaccination, at 46.39% (45/97). The difference showed statistical significance (χ(2) = 7.40, P < 0.05).

CONCLUSIONThe incidence of JE showed seasonal and regional characteristics, there were differences among ages and occupations. The difference was consistent with the difference in viral antibody level among health people in Henan province and Luoyang city.

Adolescent ; Adult ; Antibodies, Viral ; blood ; Case-Control Studies ; Child ; Child, Preschool ; China ; epidemiology ; Encephalitis Virus, Japanese ; immunology ; Encephalitis, Japanese ; blood ; epidemiology ; immunology ; Humans ; Incidence ; Infant ; Middle Aged ; Young Adult