Objective:To evaluate the role of brain-derived neurotrophic factor-antisense long-chain non-coding RNA (BDNF-AS) in amygdala in the development of neuropathic pain (NP) in rats.Methods:Healthy clean-grade male Sprague-Dawley rats, aged 2 months, weighing 200-260 g, were used to develop NP model via ligation of left L 5-6 spinal nerve, while control group was only subjected to the exposure of L 5-6 spinal nerve without ligation.This study was performed in two parts.Experiment Ⅰ Fifty-six rats were divided into 3 groups by the random number table method: sham operation group (Sham group, n=8), NP group ( n=24) and BDNF ( n=24). In BDNF group, exogenous BDNF was injected into bilateral amygdala at 1, 3, 6, 13 and 20 days after development of the model, with 100 pmol at each side.Eight rats were sacrificed at 7, 14 and 21 days after the model was developed in NP and BDNF groups and after the model was developed in Sham group, the brains were removed, and the amygdala was isolated for determination of the BDNF content (by enzyme-linked immunosorbent assay), the number of BDNF-positive cells (by immunohistochemistry), and expression of BDNF-AS (by real-time quantitative polymerase chain reaction). Experiment Ⅱ Thirty-two rats were divided into 4 groups ( n=8 each) using the random number table method: Sham operation group, NP group, BDNF group and siRNA group.At 1, 3, 6, 13 and 20 days after development of the model, exogenous BDNF 100 pmol and siRNA-BDNF-AS 50 nmol were injected into the amygdala at each side in BDNF group and siRNA group, respectively.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before development of the model (T 0) and at 4, 7, 14 and 21 days after development of the model (T 1-4). After the last behavioral test was completed, the rats were sacrificed, and the spinal cord tissues were collected to measure the contents of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α). Results:Experiment Ⅰ Compared with Sham group, the content of BDNF and the number of BDNF positive cells were significantly decreased, and the expression of BDNF-AS was up-regulated at each time point after development of the model in group NP ( P<0.05). Compared with NP group, the content of BDNF and the number of BDNF positive cells were significantly increased, and the expression of BDNF-AS was down-regulated at each time point after development of the model in group NP ( P<0.05). Experiment Ⅱ Compared with Sham group, MWT was significantly decreased and TWL was shortened at T 1-4, and the contents of IL-1β, IL-6 and TNF-α were increased in NP, BDNF and siRNA groups ( P<0.05). Compared with NP group, MWT was significantly increased and TWL was prolonged at T 1-4, and the contents of IL-1β, IL-6 and TNF-α were decreased in BDNF and siRNA groups ( P<0.05). Conclusions:The mechanism underlying the development of NP may be related to the up-regulation of BDNF-AS expression in amygdala, inhibition of BDNF synthesis and promotion of inflammatory responses in the spinal cord of rats.

Objective:To evaluate the effect of astragaloside IV on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in substantia nigra of mice with Parkinson′s disease.Methods:Forty-five SPF healthy male C57BL/6 mice, aged 8 weeks, weighing 19-25 g, were divided into 3 groups ( n=15 each) using a random number table method: control group (group C), Parkinson′s disease group (group PD) and astragaloside IV group (group A). The mouse model of Parkinson′s disease was developed by intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) 30 mg/kg everyday for 7 consecutive days.Astragaloside 20 mg/kg was intraperitoneally injected everyday at 30 min before MPTP injection for 7 consecutive days before the model was prepared in group A, and the equal volume of normal saline was given instead in group C. Behavior was measured at 1 day interval after completion of administration.The mice were then sacrificed, and the substantia nigra of the brain tissue were obtained for determination of the expression of tyrosine hydroxylase(TH), phosphorylated PI3K(p-PI3K), phosphorylated Akt(p-Akt), glial fibrillary acidic protein (GFAP) and brain-derived neurotrophic factor (BDNF) (by Western blot). Results:Compared with C group, the total distance of movement and latency of falling were significantly shortened, the hanging score was decreased, the step width was increased, the expression of TH, p-PI3K, p-Akt and BDNF in substantia nigra was down-regulated, and the expression of GFAP was up-regulated in PD group and A group ( P<0.05). Compared with PD group, the total distance of movement and the latency to fall were significantly prolonged, the hanging score was increased, the step width was reduced, and the expression of TH, p-PI3K, p-Akt and BDNF in the substantia nigra was up-regulated, and the expression of GFAP was down-regulated in group A ( P<0.05). Conclusions:The mechanism by which astragaloside IV improves motor dysfunction is related to the activation of PI3K/Akt signaling pathway, up-regulation of BDNF expression and inhibition of astrocyte activation in mice with Parkinson′s disease.

Objective:To follow up and observe the intellectual development of school children aged 8 to 10 in Baicheng County, Xinjiang Uygur Autonomous Region, which was a severe iodine deficiency disorders region in history, before and after effective control of the disease, in order to evaluate the impact of iodine supplementation on protection of children's intellectual development and provide a theoretical basis for scientific supplements of iodine.Methods:From 1989 to 2018, Combined Raven's Test for Rural in China (CRT-RC) was used to observe the intellectual development status of 660 Uyghur school children aged 8 to 10 in Baicheng County in 1989, 2002, 2006, 2012 and 2018, respectively. Children's intelligence quotient (IQ) was calculated using CRT-RC's 1987 normal sample of rural children in the same age group; the data of average iodized salt coverage rate (C-IS) and childhood goiter rate (GR) from multiple local surveys and the median urinary iodine (MUIC) of children were collected, combined with the "Criteria for Elimination of Iodine Deficiency Disorders" (GB 16006-2008) and the United Nations International Children's Emergency Fund (UNICEF) recommended standards, the status of iodine deficiency during children's growth (IDG) was divided into complete exposure to iodine deficiency, no exposure, and semi-exposed. The Flynn effect (FE) gain was calculated using norm samples of children aged 8 to 10 in 1987, 1996 and 2006 of CRT-RC, and the differences in children's intellectual development after FE correction before and after IDG reached the standard were compared.Results:The IQ of children were (81.67 ± 14.13), (83.26 ± 14.05), (89.68 ± 13.58), (98.50 ± 14.33) and (103.23 ± 15.25) points in Baicheng County in 1989, 2002, 2006, 2012 and 2018, respectively, the difference between different years was statistically significant ( F = 58.357, P < 0.01). The three indicators of C-IS, GR, and MUIC didn't meet the standards during the IDG evaluation period in the 1989, 2002, and 2006 groups, which were the complete exposure to iodine deficiency; in the 2012 group, only the MUIC met the standard, which was semi-exposed; in the 2018 group, three indicators all met the standard, which was no exposure. The FE gains of 1987 with 1996, 1996 with 2006 were 0.96 points/year and 0.74 points/year, respectively; after FE correction, the actual gains of IQ of 2002 and 2006 compared with 1989, 2012 and 2018 compared with 2006 were - 9.57, - 6.11, 4.38, and 4.67 points, respectively. Conclusions:In iodine deficiency areas, intermittent iodine supplementation (1989 - 2009) for children exposed to iodine deficiency during growth still cannot effectively protect children's intellectual development; continuous and effective iodine supplementation (2010 - 2018) with iodized salt as the core and covering children's growth period has obvious positive effects on protection of children's normal intellectual development. In the future, we will continue to observe the influence of IDG full-cycle suitable iodine nutrition on children's intellectual development.

Objective:To explore the genetic causes of oocyte maturation arrest.Methods:A pair of sisters with primary infertility was collected in the Reproductive Medicine Center of Changhai Hospital, Naval Medical University in 2018. Two PATL2 mutations were found in the patients by second-generation whole genome sequencing. Then, for all members of the family and a healthy control person, the regions near the two mutation sites were amplified and sequenced by Sanger sequencing to verify the two mutations. Furthermore, the pathogenesis of the disease was explored through bioinformatics analysis. Results:Compound heterozygous mutations of PATL2 were identified in the two sisters: a missense mutation (p.V260M) and a splicing mutation (p.R75Vfs*21). Both mutations have been reported, and they both cause abnormal splicing of PATL2 mRNA, leading to abnormal gene function. Conclusion:Our study confirmed the correlation between PATL2 mutations and human oocyte maturation arrest, and it provided molecular biomarkers for oocyte and embryo quality assessment.

Objective:To explore the genetic causes of oocyte maturation arrest.Methods:A pair of sisters with primary infertility was collected in the Reproductive Medicine Center of Changhai Hospital, Naval Medical University in 2018. Two PATL2 mutations were found in the patients by second-generation whole genome sequencing. Then, for all members of the family and a healthy control person, the regions near the two mutation sites were amplified and sequenced by Sanger sequencing to verify the two mutations. Furthermore, the pathogenesis of the disease was explored through bioinformatics analysis. Results:Compound heterozygous mutations of PATL2 were identified in the two sisters: a missense mutation (p.V260M) and a splicing mutation (p.R75Vfs*21). Both mutations have been reported, and they both cause abnormal splicing of PATL2 mRNA, leading to abnormal gene function. Conclusion:Our study confirmed the correlation between PATL2 mutations and human oocyte maturation arrest, and it provided molecular biomarkers for oocyte and embryo quality assessment.

Objective To investigate the role of miR-208b-3p in dexmedetomidine(DEX) alleviating rat myocardial ischemia reperfusion injury(MIRI).Methods Eighty adult male Wister rats were divided into the sham operation(Sham) group,I/R group,DEX group and miR-208b-3p+ DEX group.In the Sham group,a 6-0 silk suture was only placed without ligation,and other 3 groups were treated by I/R model.Before ischemia reperfusion,the DEX group received a loading dose of DEX(5 μg/kg) via the femoral vein followed by a continuous infusion of 5 μg · kg-1 · h-1 for 1 h.In the miR-208b-3p+ DEX group,rats received intramuscular injection of miR-208b-3p analogue at 24 h before ischemia reperfusion.The cardiac function indexes were monitored at 120 min after reperfusion,including heart rate(HR),left ventricular systolic blood pressure(LVSP),left ventricular end diastolic pressure(LVEDP) and left ventricular rate maximum(dp/dtmax).The serum levels of cTn-Ⅰ and CK-MB were detected and the apoptosis rate(AI) was measured by in situ apoptosis assay.The levels of oxidative stress related indicators were detected and Western blot was used to detect the level of myocardial apoptosis protein.Results Compared with the Sham group,the cardiac function in the I/R group was decreased,but the levels of CTn-Ⅰ,CK-MB,AI,MDA,Bax and Caspase-3 were increased(P＜0.05);compared with the I/R group,the above indexes in the DEX group were improved(P＜0.05);compared with the DEX group,the above indexes in the miR-208b-3p+DEX group were deteriorated(P＜0.05).Conclusion Overexpression of miR-208b-3p can eliminate the protective effect of DEX on MIRI.

Phenylketonuria (PKU) is a newborn inherited metabolic disorder caused by the genetic deficiency of hepatic enzyme phenylalanine hydroxylase (PAH) which thus in metabolic disorder of phenylalanine. In this study, ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was used to analyze the accurate amount of coenzyme Q10 (CoQ10) and the relative amount of CoQ9 in newborn dried blood spot (DBS) collected from 5 PKU newborns (3 and 30 days after birth, respectively) and 20 healthy newborns. The content of CoQ10 was (122.1±24.9 ng/mL) and (59.0±12.0 ng/mL) in DBS from healthy newborns and PKU newborns, respectively. The relative contents of cholesterol and glucose in the DBS were determined by gas chromatography-mass spectrometry (GC-MS). In comparison with healthy newborn group, the levels of CoQ10, CoQ9, cholesterol and glucose were all significantly decreased in PKU newborns. The increased content of Phe and the decreased content of CoQ10 showed significant inverse correlation in the DBS from PKU. This study provides references for diet therapy of PKU newborns.

Objective To evaluate the relationship between neuropeptide S (NPS) in the amygdala and 5-hydroxytryptamine (5-HT) and GABA in spinal dorsal horns of rats with neuropathic pain.Methods Eighty pathogen-free healthy male Sprague-Dawley rats,weighing 200-260 g,aged 2 months,were divided into 4 groups (n =20 each) using a random number table:sham operation group (group Sham),neuropathic pain group (group NP),low dose NPS group (group L-NPS) and high dose NPS group (group H-NPS).The neuropathic pain model was established by left L5,6 spinal nerve ligation (SNL) in anesthetized rats.NPS was injected into the bilateral amygdala at 3,6,9,12,15 and 18 days after SNL in LNPS group (10 pmol per side) and H-NPS group (100 pmol per side).The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 2 days before SNL and 1,4,7,11,14,17 and 21 days after SNL.Five rats were selected at 7,14 and 21 days after SNL and sacrificed,and the lumbar segment (L5) of the spinal cord was removed for detection of the expression of 5-HT and GABA in spinal dorsal horns by immunofluorescence histochemistry.Results Compared with group Sham,the MWT was significantly decreased,the TWL was shortened,and the expression of 5-HT and GABA in spinal dorsal horns was down-regulated in NP,L-NPS and H-NPS groups (P＜0.05).Compared with group NP,the MWT was significantly increased,the TWL was prolonged,and the expression of 5-HT and GABA in spinal dorsal horns was up-regulated in L-NPS and H-NPS groups (P＜0.05).Compared with group L-NPS,the MWT was significantly increased,the TWL was prolonged,and the expression of 5-HT and GABA in spinal dorsal horns was up-regulated in group H-NPS (P＜0.05).Conclusion The spinal mechanism of endogenous analgesia induced by NPS in the amygdala may be related to up-regulation of the expression of 5-HT and GABA in spinal dorsal horns of rats with neuropathic pain.

Objective]To summarize Professor Chen Huade’s clinical experience on treatment of hyperactivity of liver yang type vertigo by acupuncture. [Method]By combining with expositions of past dynasties, collecting and analysis of the clinical cases of liver yang type vertigo, to summarize the etiology and pathogenesis of liver yang type vertigo and Professor Chen’s treatment characteristics, and take a case as evidence.[Result]In Professor Chen’s opinion, the basic pathogenesis of hyperactivity of liver yang type vertigo is hepatorenal Yin deficient liver Yang syndrone. Belong to excess in the upper and deficiency in the lower, asthenia in origin and asthenia in superficiality. Based on the theory of “treating disease from the root”,cure this disease by the body acupuncture plus eartip bloodletting therapy, each had a good curative effect.The evidence showed, after the body acupuncture plus eartip bloodletting therapy ten times, vertigo symptom relieved obviously, blood pressure dropped to normal range. In the following up of 3 months, none had a recurrence. [Conclusion]Because of its cheap, non-toxic side effects, significant efficacy, and the characteristics of both symptoms and root causes curing, it is gradually accepted by more and more patients.Professor Chen Huade’s treatment on hyperactivity of liver yang type vertigo by acupuncture has significant effect in clinic which is worth spreading.

Objective To systematically assess the clinical efficacy and safety of acupuncture treatment for vertigo with excess syndrome. Methods Chinese and English literature about acupuncture treatment of vertigo with excess syndrome published in recent years was comprehensively searched. The quality of the retrieved literature meeting the inclusion criteria of randomized controlled trial was assessed and its data was collected. A Meta analysis of the included studies was carried out.Results Finally, 10 articles with randomized controlled trials containing a total of 688 patients were included in the analysis. The included literature was assessed at lower quality using Cochrane evaluation member manual 5.1. A Meta analysis showed that the efficacy rate of acupuncture treatment for vertigo with excess syndrome was higher than that of Western drugs alone [M-H OR 4.84, 95%CI (2.39, 9.81),P<0.0001]. Combined acupuncture and Chinese herbal medicine was superior to Chinese herbal medicine alone [M-H OR 3.82, 95%CI (2.06, 7.10),P<0.0001]. Vertigo symptom and function scoring showed day 3 of treatment [MD 4.66, 95%CI (2.97, 6.35)], day 7 of treatment [MD 0.95, 95%CI (0.03, 1.86)] and day 14 of treatment [MD 0.89, 95%CI (0.71, 2.49)]. There were statistically significant differences in the vertigo symptom and function scores between the two groups of patients at day 3 and 7 of treatment. There were no statistically significant differences in the scores between the two groups of patients at day 14 of treatment. Conclusions Acupuncture or combined acupuncture and Chinese herbal medicine are effective and highly safe in treating vertigo with excess syndrome, but high-quality, multi-center and large-sample RCT studies still need to be conducted for validation and support.