Objective To evaluate the effects of epidural capsaicin on pain threshold and nerve tissue structure in adult rats. Methods Twenty-four adult male Wistar rats weighing 250-300 g were used in this study. A catheter was inserted into epidural space at L5,6 according to the method of Philippe. 0.4 ml of capsaicin 0.1% (group A), 0.25% (group B) or 0.5% (group C) or 10% Tween 80 (control group) was injected via the epidural catheter. Pain threshold was measured by thermal stimulation of the tail before (baseline) and on the 3rd, 7th and 14th day after epidural capsaicin. The animals were then killed. The lumbar segment (L4-6) of the spinal cord and spinal nerve roots were removed immediately for light and electron microscopic examination. Results Pain threshold was significantly higher in group A, B and C than in control group (P

AIM: To construct recombinant lentiviral vector with short hairpin RNA (shRNA) of CREB gene, and to investigate the effect of CREB gene silencing on mitochondrial morphology and cell apoptosis in oxygen-glucose deprivation/reoxygenation (OGD/R)-induced cortical neurons.METHODS: Three lentiviral vectors pLentiLox3.7 (PLL) inserted shRNA fragments targeting CREB gene were co-transfected with the packaging plasmids psPAX2 and pMD2.G to the 293T cells, and the virus particles, which was infected with the primary cortical neurons, was encapsulated.The protein expression of CREB was detected by Western blot.The mitochondrial morphology, cell apoptosis and the expression of Bcl-2 and Bax were evaluated by the methods of MitoTracker red, TUNEL and Western blot in OGD/R induced cortical neurons after CREB gene silencing.RESULTS: The pLL-CREB-shRNA1 was the most effective shRNA, which inhibited 80% CREB gene expression in the cortical neurons.The mitochondrial was appeared dot and fragment morphology in OGD/R induced cortical neurons with transfected pLL-CREB-shRNA1 plasmid.In addition, the expression of Bcl-2 was decreased, the expression of Bax, and the apoptosis of the neurons were increased by tranfected with pLL-CREB-shRNA1.CONCLUSION: CREB shRNA recombinant lentiviral vector specifically inhibits the expression of CREB gene.CREB gene silencing promotes the cell apoptosis and mitochondrial morphological changes in the cortical neurons induced by OGD/R.

Objective To investigate the relationship between Kashin-Beck Disease(KBD) and Human Parvovirus B19(HPVB19).Methods HPVB19DNA was detected in 55 sera of KBD patients and 52 healthy in adjacent non-endemic area and 35 healthy sera in normal area using PCR and then linked the HPVB19DNA to pGEM-T vector.The nucleotide sequence was analyzed and compared with HPVB19 nucleotide sequence published by Genebank and another in Journal of virology.Results HPVB19DNA was found in 16 of 55 sera in KBD patients,and the HPVB19DNA position rate(29.09%) is significantly higher than that of the two healthy control groups(11.54%、11.42% respectively)(P＜0.05).The nucleotide sequence homologies compared with the two published nucleotide sequence were 97.75%、97%,respectively.The putative amino acid homologies compared with the tow published were 93.5%.The amino acid variation was greater than the nucleotide sequence variation because of a base insertion.Conclusion There was a close relationship between HPVB19 infection and Kashin-Beck Disease.

Objective:To summarize the diagnosis features of the prenatal genetic diagnosis of fetal renal cystic disease and to explore the clinical feasibility and significance of prenatal genetic diagnosis of congenital cystic nephrosis.Methods:A total of 25 fetuses with congenital renal cystic disease were examined via invasive prenatal diagnosis in Henan Provincial People's Hospital from June 2017 to September 2019. Amniotic fluid samples were extracted by amniocentesis. Chromosomal microarray analysis (CMA) were performed in 17 cases. In addition to CMA, the other 8 cases were analyzed by G-band karyotype. Whole exome sequencing (WES) was performed in 6 cases which got normal results by CMA and karyotype, and highly suspected as hereditary disease.Results:Of the 25 fetuses assessed, 4 cases (16.0%) pathogenic copy number variation (pCNV) were found, including 2 cases of 17q12 deletion, 1 case of 10p15.1p14 deletion and 1 case of 4q21.28q22.1 deletion(including PKD2 gene). There were 8 cases without chromosome abnormality by karyotype analysis. Six clinical WES analysis found NPHS1 gene c.1440+1 G>A and c.925G > T mutations were related to Finnish type congenital nephrotic syndrome in 1 case, PKD1 gene c.6878C>T mutation was related to autosomal dominant polycystic kidney disease (ADPKD) in 1 case, and there was no definitive mutation in 4 cases. Conclusions:CMA and next generation sequencing are powerful tools for accurate diagnosis, treatment and genetic counseling of fetal congenital renal cystic diseases. For congenital cystic nephropathy, genetic detection is helpful to clarify the etiology, and provide more exactly informations for prognosis evaluation, treatment and family genetic counseling.

Objective To investigate the endothelial dysfunction in pre-hypertension and its influencing factors.Methods A total of 373 youth were divided as the subjects into hypertension group (HBP group),prehypertension group (PHT group) and normal blood pressure group (NBP group).Endothelial function was assessed based on carotid intima-media thickness (IMT),brachial artery flow-mediated dilation (FMD) and brachial-ankle pulse wave velocity (baPWV).Results IMT and baPWV in PHT group were higher than those in NBP group (P＜0.05),but did not reach the significant difference when compared with HBP group (P＞0.05).Compared with HBP,the levels of FMD in PHT group significantly increased (P＜ 0.05);however,no difference was observed in comparison with NBP group (P＞0.05).In the early stage of hypertension,diastolic BP (β=-0.120,P＜0.05) and body mass index (β=-0.115,P＜0.05) were negatively correlated with FMD;diastolic BP (β=0.146,P＜0.05),2-hour glucose (β=0.147,P＜0.05),high-density lipoprotein cholestrol (β=0.150,P＜0.05),and waist-hip ratio (β=0.126,P＜0.05) showed a positive correlation with IMT.baPWV was correlated with systolicBP (β=0.358,P＜0.01),waist circumference (β=0.254,P＜0.05),fasting glucose (β=0.155,P＜0.05),postprandial 2 h blood glucose (β =0.152,P ＜0.05),uric acid (β =0.206,P ＜ 0.05),and C-reactive protein (β=0.099,P＜0.05).Corclusion Our study shows that endothelial dysfunction may exist in the prehypertensive young,and several cardiovascular risks contribute to its development in the early stage of hypertension.

Objective To investigate prospectively the diagnostic significance of ulnar/median nerve amplitude ratio in motor neuron disorders.Methods Patients referral to our department between May 2009 and February 2012,due to muscle weakness,inflexible,cramps and atrophy,were consecutively enrolled.Conventional nerve conduction studies of 4 extremities (using surface electrodes) and needle electromyography were performed in all patients with fixed examiner.The compound muscle action potentials (CMAP) were recorded from abductor digiti minimi (ADM) and abductor pollicis brevis (APB) with stimulation of ulnar and median nerve at wrist,respectively.Moreover,the ratio of CMAP amplitude between ADM and APB (ADM/APB) was calculated in the patients who met the diagnostic criteria for definite amyotrophic lateral sclerosis (ALS) and Hirayama disease (HD).The patients with Guillain-Barré syndrome (GBS) and other popyneuropathies (PN) were served as case-controls,and 34 healthy volunteers (aged (45.7 ± 16.3) years) as normal-controls.Results (1) There were 78 cases with ALS,25 HD,51 GBS and 140 other PN,with the mean age(years) of 54.7 ± 11.6,17.6 ± 2.2,41.3 ± 18.4 and 57.1 ± 14.3,respectively.(2) ADM/APB in the ALS subgroup was 2.28 ±2.87 (0.12-22.38),HD0.66±0.36 (0.05-1.34),GBS 1.42 ± 1.33 (0.25-9.85),other PN 1.36 ± 1.48 (0.08-14.44) and normal-controls 1.07 ± 0.28 (0.61-1.64,F =6.872,P =0.000),respectively.(3) The areas under receiver operator characteristic curve in patients with ALS was 0.830 (s-x =0.039) and HD 0.691 (sx =0.039,P =0.000) ; the diagnostic sensitivity and specificity for ALS patients were 36.7％ and 93.3％,respectively,with cutoff value of ADM/APB =2; and the diagnostic sensitivity and specificity for HD patients were 53.6％ and 89.0％,respectively,with cutoff value of ADM/APB =0.7.Conclusions The ulnar/median CMAP amplitude ratio increases in ALS,but decreases in HD,which may be served as a relatively specific electrophysiological index.ADM/APB amplitude ratio ＞ 2.0 is suggested to be a diagnostic parameter for ALS and ＜ 0.7 for HD.

Objective To investigate the involvement of posterior auricular muscle (PAM) and the effect of botulinum toxin type A (BTX-A) injection into PAM in patients with hemifacial spasm (HFS)complicated by auricular symptoms.Methods Sixty-three consecutive HFS patients with auricular symptoms such as tinnitus or murmur,"ticking" or a "clicking" sound and discomfort on the same side,referred to our department between July,2009 and January,2010,were enrolled,and the diagnosis of idiopathic HFS was clinically made.The patients were largely randomized into two groups according to the order of referral.One was regular group including 33 cases whose injection sites were routinely at the frontal,orbicularis oculi,zygomaticus and buccinator muscles while another was PAM group including 30 cases,in which 4 units of BTX-A was additionally injected to the PAM.Before and after injection,the test of blink reflex was performed and lateral spread of blink reflex to orbicularis oris(OO)and PAM,I.e.Abnormal muscle response(AMR),were recorded,and the peak-peak amplitude of AMR was measured.The patients were followed up clinically and electrophysiologically at least 4 weeks((29.5±2.5)days) later.Results(1)The patients reported that their auricular symptoms subsided after injection in both groups.The remission rate was 45.5%(15/33)in the regular group and 76.7%(23/30)in the PAM group,respectively,with a higher rate in the PAM group(x2=6.40,P=0.011).(2)In both groups the AMR amplitude decreased significantly after injection.In the regular group,the OO amplitudes (μV) before and after injection were 304.0±30.3 and 129.3±9.6(t =5.820,P =0.000),and PAM amplitudes,298.0±33.3 and 184.7±20.2(t=2.818,P=0.014),respectively.In the PAM group,OO amplitudes were 405.3±66.7 and 116.0±10.0(t=4.214,P=0.001),PAM amplitudes,390.0±53.6 and 72.0±9.7(t=6.011,P=0.000),respectively.(3)The decrease of PAM amplitudes in the PAM group was more significant compared with those in the regular group (t=4.237,P=0.001).Conclusions In HFS patients with auricular symptoms,the electrophysiological studies are helpful for the guidance of treatment;and the auricular symptoms could be better improved after BTX-A injection into PAM in addition to those regular injection sites.

Objective:To improve the quality standard for Qingyuantiaozhi capsules. Methods:The main components of the prep-aration, such as Chrysanthemum, Anthraquinones, Hawthorn and Radix Rehmanniae Preparata, were identified by TLC qualitatively. The content of chlorogenic acid in chrysanthemum was determined by HPLC. A DIKMA Spursil C18(250 ×4.6 mm,5 μm)column was used with methanol-0. 2% phosphoric acid solution(9 ∶91) as the mobile phase. The flow rate was 1. 0 ml·min-1, the detection wavelength was set at 327 mn and the sample size was 20 μl. Results:The spots in TLC were clear without any interference. The cali-bration curve was linear within the range of 4. 425 2-30. 976 4μg·ml-1(r=0. 999 9) for chlorogenic acid. The average recovery was 101. 18% (RSD=1. 88%, n=6). Conclusion:The improved quality standard is specific, accurate and reproducible, which can be used for the quality control of Qingyuantiaozhi capsules.

Objective To evaluate the effect of tripterine on hydrochloric acid-induced acute lung injury(ALI)in mice.Methods Eighteen pathogen-free healthy adult male ICR mice,aged 7-9 weeks,weighing 25-30 g,were divided into 3 groups(n=6 each)using a random number table:control group(group C),hydrochloric acid-induced ALI group(group ALI)and tripterine group(group T).ALI was induced by a single intratracheal instillation of hydrochloric acid 2 ml/kg(pH 1.5)via a 24-gauge angiocatheter inserted into the trachea in pentobarbital sodium-anesthetized mice.Tripterine 3 mg/kg was injected intraperitoneally once a day for 3 consecutive days,and then the model was established in group T.The mice were sacrificed at 6 h after instillation,and lung specimens were obtained for microscopic examination and for determination of the levels of tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),macrophage migration inhibitory factor(MIF)and myeloperoxidase(MPO)in lung tissues.Results Compared with group C,the levels of TNF-α,IL-6,MIF and MPO were significantly increased at 6 h after instillation in ALI and T groups(P<0.01).Compared with group ALI,the levels of TNF-α,IL-6,MIF and MPO were significantly decreased at 6 h after instillation in group T(P<0.01).The pathological changes of lung tissues were significantly attenuated in group T compared with group ALI.Conclusion Tripterine can attenuate hydrochloric acid-induced ALI in mice.

BackgroundMonocyte chemotactic protein-1 (MCP-1)plays an important role in the tumor,inflammation,diabetic retinopathy and other neovascular disease,but the expression and the role of MCP-1 in the oxygen induced retinopathy(OIR) model have rarely been reported. Objective This study was to investigate the expression of MCP-1 in the retina development of newborn mouse and in mouse models with OIR.Methods C57BL/6J newborn mice were divided into two groups and 60 mice in each group.Mice in OIR group were exposed to 75％ oxygen for 5 days and then to room air.All mice in normal control group exposed to room air only.Ten mice in each group were randomly chosen and sacrificed at postnatal 5,7,12,14,17,21 days.The expression of MCP-1 in mouse retina was detected with the method of immunohistoehemistry and reverse transcription polymerase chain reaction(RT-PCR).Results MCP-1 positive cells were seen in normal mouse retina.Up-regulation of MCP-1 positive cells was detected both in 12 days in normal control group and in 14 days in OIR group.MCP-1 mRNA was detected in mouse retina at 5 days,and a transient up-regulation of MCP-1 mRNA was observed in 12 days in normal control group.MCP-1 mRNA in OIR group significantly increased in 14 days in comparison with the normal control group( P =0.028,P =0.001 ). Conclusions Expression of MCP-1 is detectable in whole retinal development procession of mice.A transient up-regulation of MCP-1 expression is detected in the critical period of retinal vascular development in mice models with OIR,which is closely related to the retinal vascular development and progression of retinal new vessels.