Pulsed electromagnetic fields stimulation combined with sclerostin antibody improves bone metabolism and bone microstructure in postmenopausal osteoporosis rabbits / 中国组织工程研究

BACKGROUND: Both pulsed electromagnetic fields (PEMF) stimulation and sclerostin antibody (Scl-Ab) have good effects on the bone metabolism of ovariectomized (OVX) New Zealand rabbits, but research on the combined intervention of PEMF and Scl-Ab in the OVX rabbits is rarely reported. OBJECTIVE: To explore the effect of PEMF combined with Scl-Ab on postmenopausal osteoporosis and to explore the therapeutic value for osteoporosis. METHODS: An animal model of postmenopausal osteoporosis was made in New Zealand white rabbits after ovariectomy. The experimental animals were randomly divided into OVX control group, PEMF group, Scl-Ab group and PEMF+Scl-Ab group, with 10 rats in each group. On the 1st day after surgery, the PEMF group was given PEMF magnetic therapy once a day; the Scl-Ab group was given subcutaneous injection of Scl-Ab twice a week; the PEMF+Scl-Ab group received PEMF magnetic therapy once a day, five times a week, and Scl-Ab subcutaneous injections twice a week; the OVX group was injected subcutaneously with the same dose of normal saline twice a week for 8 weeks. After 8 weeks of treatment, bone metabolism index, bone mineral density, and MicroCT bone microstructure parameters were detected. All animal procedures were approved by the Department of Experimental Animal Science, Fudan University (approval No. 20171263A193). RESULTS AND CONCLUSION: Bone mineral density was significantly decreased in the New Zealand white rabbits after 6 months of OVX, suggesting that the osteoporosis model was successfully established. Compared with the OVX group, the bone mineral density of the L3 vertebral body in the PEMF group, the Scl-Ab group and the PEMF+Scl-Ab group increased significantly (P < 0.05). Compared with the OVX group, serum bone-specific alkaline phosphatase levels were significantly higher, and serum tartrate-resistant acid phosphatase 5b levels were significantly lower in the PEMF group, the Scl-Ab group and the PEMF+Scl-Ab group. The serum tartrate-resistant acid phosphatase 5b level in the PEMF+Scl-Ab group was significantly lower than that in the PEMF group and the Scl-Ab group. The bone metabolism index and bone microstructural parameters (bone volume fraction, trabecular thickness, trabecular number, and trabecular separation) of the PEMF+Scl-Ab group were significantly better than those of the PEMF group and the Scl-Ab group (all P < 0.05). These findings indicate that the combination of Scl-Ab and PEMF can enhance bone mineral density and improve bone metabolism and bone microstructure in postmenopausal osteoporosis New Zealand white rabbits.

Effect of acupoint catgut embedding therapy on joint attention and social communication in children with autism spectrum disorder: a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the clinical effect on the joint attention and social communication in children with autism spectrum disorder (ASD) between the combined treatment of acupoint catgut embedding therapy with rehabilitation training and the simple application of rehabilitation training.@*METHODS@#A total of 60 children with ASD were randomized into an observation group and a control group, 30 cases in each one. In the control group, the routine ASD rehabilitation training was adopted, including conductive education, speech training and music therapy, once a day. In the observation group, on the base of the treatment as the control group, the catgut embedding therapy was applied to Shenting (GV 24), Baihui (GV 20), Shenmen (HT 7) and the optic area (the point of scalp acupuncture), once a week. In the two groups, the treatment for 4 weeks was as one course, at the interval of 1 week between the courses. A total of 3 courses of treatment were required. The social domain of the autism treatment evaluation checklist (ATEC) and the autism behavior checklist (ABC) were adopted to assess the therapeutic effect of the two groups.@*RESULTS@#After treatment, the scores of each item of the social domain in ATEC and the scores of ABC (feeling, communication, physical movement, language and healthy behavior) were all lower than those before treatment in the two groups (<0.01). The scores of each item in the observation group were lower than those in the control group after treatment (<0.05).@*CONCLUSION@#The combined treatment of acupoint catgut embedding therapy with the rehabilitation training effectively improves in the joint attention and social communication. The therapeutic effect of this combined treatment is better than the simple application of rehabilitation training.

Dynamic simulation analysis on gait features of hemiplegic patients / 医用生物力学

Objective To study the intrinsic relationship between hemiplegic gait features and abnormal muscle strength of lower limbs,so as to elucidate the muscle strength causes of hemiplegic gait and provide recommendations for its clinical treatment.Methods Models of hemiplegic gait and normal gait were established for simulation,and the validity of the models was verified by comparing the simulation model with measured data of the normal gait.The differences in kinetic data,ground reaction force (GRF) and muscle force between the two models were analyzed to explore the different dynamic characteristic of hemiplegic gait and normal gait.Results The complex correlation coefficient between LifeMOD simulation results and measured data was 0.922,indicating that the established dynamic model was reasonable and effective.Hemiplegic patient with low tibialis anterior muscle strength led to ankle dorsiflexion inadequacy during initial ground period,and low gastrocnemius muscle could not achieve the promoting effect from ground during preswing period.Conclusions The strength weakness of tibialis anterior muscle and gastrocnemius are the main reasons for foot drooping and other hemiplegic gait characteristics.LifeMOD modeling and simulation can assist the diagnosis of abnormal muscle strength in hemiplegia patients.

Dynamic simulation analysis on gait features of hemiplegic patients / 医用生物力学

Objective To study the intrinsic relationship between hemiplegic gait features and abnormal muscle strength of lower limbs, so as to elucidate the muscle strength causes of hemiplegic gait and provide recommendations for its clinical treatment. Methods Models of hemiplegic gait and normal gait were established for simulation, and the validity of the models was verified by comparing the simulation model with measured data of the normal gait. The differences in kinetic data, ground reaction force (GRF) and muscle force between the two models were analyzed to explore the different dynamic characteristic of hemiplegic gait and normal gait. Results The complex correlation coefficient between LifeMOD simulation results and measured data was 0.922, indicating that the established dynamic model was reasonable and effective. Hemiplegic patient with low tibialis anterior muscle strength led to ankle dorsiflexion inadequacy during initial ground period, and low gastrocnemius muscle could not achieve the promoting effect from ground during preswing period. Conclusions The strength weakness of tibialis anterior muscle and gastrocnemius are the main reasons for foot drooping and other hemiplegic gait characteristics. LifeMOD modeling and simulation can assist the diagnosis of abnormal muscle strength in hemiplegia patients.

IVS8+1 DelG, a Novel Splice Site Mutation Causing DFNA5 Deafness in a Chinese Family / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Nonsyndromic hearing loss (NSHL) is highly heterogeneous, in which more than 90 causative genes have currently been identified. DFNA5 is one of the deafness genes that known to cause autosomal dominant NSHL. Until date, only five DFNA5 mutations have been described in eight families worldwide. In this study, we reported the identification of a novel pathogenic mutation causing DFNA5 deafness in a five-generation Chinese family.</p><p><b>METHODS</b>After detailed clinical evaluations of this family, the genomic DNA of three affected individuals was selected for targeted exome sequencing of 101 known deafness genes, as well as mitochondrial DNA and microRNA regions. Co-segregation analysis between the hearing loss and the candidate variant was confirmed in available family members by direct polymerase chain reaction (PCR)-Sanger sequencing. Real-time PCR (RT-PCR) was performed to investigate the potential effect of the pathogenic mutation on messenger RNA splicing.</p><p><b>RESULTS</b>Clinical evaluations revealed a similar deafness phenotype in this family to that of previously reported DFNA5 families with autosomal dominant, late-onset hearing loss. Molecular analysis identified a novel splice site mutation in DFNA5 intron 8 (IVS8+1 delG). The mutation segregated with the hearing loss of the family and was absent in 120 unrelated control DNA samples of Chinese origin. RT-PCR showed skipping of exon 8 in the mutant transcript.</p><p><b>CONCLUSIONS</b>We identified a novel DFNA5 mutation IVS8+1 delG in a Chinese family which led to skipping of exon 8. This is the sixth DFNA5 mutation relates to hearing loss and the second one in DFNA5 intron 8. Our findings provide further support to the hypothesis that the DFNA5-associated hearing loss represents a mechanism of gain-of-function.</p>

Summary of software OpenSim with focus on its human motion modeling theory and application field / 医用生物力学

The current experimental methods, such as the motion capture system, high-speed video, plane photography and surface EMG, cannot show the details of the muscle strength, or explain the nerve-muscle coordination mechanism during human motion. Thus, computer modeling and simulation are developed and become a new method for researchers in China to study human motion, and the software LifeMOD, AnyBody, ANSYS are mainly used even if these softwares have some limitations such as less precise muscle control and relatively higher prices. Developed by Stanford University, OpenSim is an open-source platform for muscle model development, simulation and analysis of neuromuscular system based on C++ and Java. In this paper, on the basis of computer modeling and mathematics theoretical derivation, the OpenSim modeling theory and simulation steps were introduced in detail to provide some theoretical references for researches on sport biomechanics. The future research directions by using OpenSim modeling simulation to study neuromuscular mechanism of the hemiplegic gait and Parkinson gait, and to improve and perfect the rehabilitation ways of the abnormal gait were also discussed.

Risk factors for interictal epileptiform discharges on electroencephalogram in children with spastic hemiplegic cerebral palsy / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical symptoms and features of interictal epileptiform discharges (IED) on electroencephalogram (EEG) in children with spastic hemiplegic cerebral palsy (CP) and to analyze the risk factors for IED.</p><p><b>METHODS</b>Eighty-three children with spastic hemiplegic CP were recruited, and their clinical data, results of video-electroencephalogram, imaging findings, and cognitive levels were collected. The influencing factors for IED were determined by multiple logistic regression analysis.</p><p><b>RESULTS</b>The incidence of epilepsy was 13% in children with spastic hemiplegic CP; 34% of these cases had IED. The incidence of epilepsy in children with IED (32%) was significantly higher than that in those without IED (4%) (P<0.01). The incidence of IED in children with complications and brain cortex impairment increased significantly (P<0.01). The incidence of IED varied significantly between patients with different cognitive levels (P<0.01). Brain cortex impairment (OR=11.521) and low cognitive level (OR=2.238)were risk factors for IED in children with spastic hemiplegic CP (P<0.05).</p><p><b>CONCLUSIONS</b>Spastic hemiplegic CP is often found with IED on EEG, and the incidence of epilepsy is higher in children with IED than in those without IED. Brain cortex impairment and low cognitive level have predictive values for IED in children with spastic hemiplegic CP.</p>

Recipient dendritic cells modified by RNA interference targeting CD80 and CD86 elicit T cell hyporesponsiveness via enhanced T cell apoptosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Despite extensive research, the mechanism of immature dendritic cells (DCs) induced immune hyporesponsiveness remains incompletely understood.</p><p><b>METHODS</b>Recipient DCs from C3H mouse bone marrow cells were incubated with donor antigen from splenic lymphocytes of C57BL/6 mouse; these DCs were transfected with CD80/86 specific siRNA using lentiviral vectors. Flow cytometry was used to evaluate expression of CD80/86 on the antigen-pulsed recipient DCs. Immune regulatory activity was examined by mixed lymphocyte reaction, in which irradiated DCs were cultured with C3H spleen T cells. After the reaction, interleukin (IL)-2, IL-4, IL-10, and interferon (INF)-γ levels of mixed lymphocyte reaction culture supernatant were measured by enzyme-linked immunosorbent assay. The apoptotic T lymphocytes were identified by Annexin V and CD3 staining.</p><p><b>RESULTS</b>There was a significant inhibition of CD80/86 expression in DCs transfected with CD80/86 lentiviral vectors compared with the control groups (P < 0.05), indicating the specificity of RNA interference. Enzyme-linked immunosorbent assay results showed a significant reduction of INF-γ, IL-2 and IL-10 in the CD80/86 lentivirus transfected group compared to the control groups (P < 0.05). There was no significant difference in IL-4 levels between the groups (P > 0.05). We also showed that CD80/86 low DCs loaded with alloantigen (1) stimulated low T cell proliferative responses via the indirect recognition pathway and (2) enhanced apoptotic activity (P < 0.05) in co-cultured T cells.</p><p><b>CONCLUSIONS</b>Lentiviral vector transfection can effectively and specifically knock down target genes in DCs. The CD80/86 low DCs may show tolerogenic activity via induction of T-cell apoptosis, thereby modulating the activity of recipient-derived DCs. The use of this approach may potentially be clinically applicable.</p>

Clinical observation on treatment of clearing the Governor Vessel and refreshing the mind needling in neural development and remediation of children with cerebral palsy / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of clearing the Governor Vessel and refreshing the mind needling in neural development and remediation of children with cerebral palsy.</p><p><b>METHODS</b>A total of 200 cases of children with cerebral palsy were randomly assigned to the treatment group (100 patients) and the control group (100 patients). The treatment group was given the combined therapy of acupuncture and rehabilitation training, and the chosen acupoints were 13 points of the Governor Vessel, Shenshu (BL 23), Taixi (KI 3), Yanglingquan (GB 34), Zusanli (ST 36) and Sanyinjiao (SP 6), and points of refreshing the mind were also selected, which included puncturing Shenting (GV 24) toward Qianding (GV 21), puncturing Qianding (GV 21) toward Baihui (GV 20), puncturing Baihui (GV 20) toward Naohu (GV 17) and Sishencong (Ex-HN 1). The control group was only treated with rehabilitation training. A contrastive analysis of the therapeutic effect between acupuncture combined with rehabilitation training and rehabilitation training alone was made after a treatment course of 3 months. The Gross Motor Function Measure (GMFM) and Beijing Gesell Developmental Scale were adopted to assess the neural development and rehabilitation outcomes of the two groups. In addition, skull CT/MRI was adopted to evaluate the plerosis of injured cerebral nerve after treatment.</p><p><b>RESULTS</b>The total effective rate in treatment group was 87% (87/100), significantly higher than the 55% (55/100) in the control group. The children's development quotient (DQ) tested by Gesell Developmental Scale and scores tested by GMFM in the treatment group were obviously higher than those in the control group (P<0.01). The improving and curing rates presented by skull CT/MRI in the treatment group were higher than those in the control group (P<0.01).</p><p><b>CONCLUSIONS</b>Clearing the Governor Vessel and refreshing the mind needling could accelerate the recovery of injured brain nerve and the reconstruction of brain function. The acupuncture therapy could ameliorate both the motor development and cognitive development. On the other hand, the forward curative effect of acupuncture combined with rehabilitation training was significantly better than the rehabilitation training alone.</p>

Simultaneous determination of catechins and alkaloids in brick-tea by high performance liquid chromatography-diode array detection / 中国地方病学杂志

Objective To develop a method for determination of catechins and alkaloids in brick-tea by high performance liquid chromatography(HPLC) with diode array detection(DAD),and to explore optimum extraction conditions for catechins and alkaloids components in brick-tea.Methods Catechins and alkaloids were separated and detected by HPLC.Orthogonal experiment and paired t test was carried out to compare the effect of water and alcohol as extraction solvents and to obtain optimum extraction conditions for extracting catechins and alkaloids components in brick-tea.Results Six kinds of catechins of catechin,epicatechin,gallocatechin,epigallocatechin,epicatechin gallate,epigallocatechin gallate and two kinds of alkaloids of caffeine and theobromine were separated and detected simultaneously.Calibration curves between peak areas and concentration of each component in bricktea were linear within a suitable concentration range,and coefficients of determination (R2) were between 0.9990-0.9999; spiked recoveries were from 83.78％ to 106.35％,and relative standard deviations(RSD) were between 0.50％-1.51％.The optimum extraction condition for catechins and alkaloids was 80％ ethanol,solid-liquid ratio of 1:10,temperature 80 ℃ and extraction time 30 min.Conclusion Alcohol as extraction solvents with optimal combination on HPLC,six kinds of catechins and two kinds of alkaloids are separated accurately,qualitatively,rapidly and sensitively.

Quality of life survey of inhabitants living in endemic arsenism areas after drinking-water improvement / 中国地方病学杂志

ObjectiveTo find out the quality of life of inhabitants living in endemic arsenism areas after drinking-water improvement and related influencing factors.MethodsCase group ( 118 people) of arsenicosis,and control group (237 people ) of nearby residents without arsenicosis in endemic arsenic areas after drinking-water improvement in Shanxi and Inner Mongolia and outside control group(116 people) not exposed to arsenic in nearby non-arsenic areas were selected to be interviewed using homogeneous validity and reliability of the World Health Organization quality of life(WHOQOL-BREF,Chinese Version) in 2010,and physical health,psychological,social relationships,environment scores and total score were calculated,respectively.Covariance analysis was used to find out the real difference in the three groups and multiple linear regression analysis was used to explore the influencing factors on quality of life.Results The scores of physical health,psychological,social relationships,environment and total score of case group were ( 12.14 ± 2.68),( 12.18 ± 2.22),( 14.81 ± 2.24),(11.33 ± 1.92),(69.27 ± 16.23) points,respectively,and those of internal control group were ( 13.39 ± 2.67),( 12.80 ± 1.88 ),( 15.16 ± 1.80),( 11.72 ± 1.62),(75.49 ± 14.50) points,respectively,those of control group were (13.91 ± 254),(13.67 ± 2.14),(15.07 ± 1.36),(12.64 ± 1.68),(78.41 ± 12.03) points,respectively.Them was significant difference in physical health,psychological,environment scores and total score among the three groups(F =7.57,10.74,13.44,10.77,all P ＜ 0.05),of which case group was significantly lower than those of the inside and outside control group(all P ＜0.01 ),while in the control group the psychological and environment scores were lower than those of the external control group(all P ＜ 0.01 ).Health,whether-or-not arsenicosis patient,annual per capita income,age and sex were influential factors of quality of life among inhabitants living in endemic arsenism areas after drinking-water improvement (all P ＜ 0.05 ).Conclusions Quality of life among inhabitants living in endemic arsenism areas after drinking-water improvement is lower than residents of non-arsenic areas.

National monitoring report of drinking-water-born endemic arsenicosis in 2010 / 中国地方病学杂志

Objective To master the implementing progress of preventive measures for control of endemic arsenicosis,to observe the dynamic changes and provide the basis for prevention of national drinking-water-born endemic arsenicosis.Methods The surveillance was strictly carried out according to “The Monitoring Project for the National Drinking-Water-Born Endemic Arsenicosis(trial implementation)” in 2010.According to the results of previous investigation,11 provinces and Xinjiang Production and Construction Corps were selected as the surveillance provinces; eighty six endemic arsenicosis villages which had exposed population over 100 persons were chosen as surveillance villages in each province; forty eight potential endemic arsenicosis villages were recognized as the monitoring villages which had over 100 exposed people too.We investigated the surveillance counties and villages about the running state of water-improving projects,the arsenic content in water from resident house in potential endemic arsenicosis villages and the survey on endemic arsenicosis status based on the residents who lived in the surveillance villages.Results ①Total of 219 water-improving projects in 32 counties were monitored and 1673 villages were covered,benefiting 69.07 million population.②Total of 134 villages with high level of water arsenic were investigated.Water quality improved villages was 86,accounting for 64.18％.Normal working projects accounted for 91.86％ (79/86),intermittentl y working accounted for 4.65％ (4/86) and abandoned projects accounted for 3.49％(3/86).Passing rate of water arsenic of the projects was 91.86％(79/86).Total of 48 villages without water-improving project were investigated,and families with excessive level of water arsenic was 35.04％ (356/1016).③Total of 20 885 persons were examined in villages with improved water,incidence of endemic arsenicosis was 4.44％ (928/20 885).Among them,patients with mild arsenicosis was 3.27％ (682/20 885),with moderate was 0.80％(167/20 885) and serious patients was 0.37％(78/20 885),and detection of skin cancer 1 person.Totally 6166 persons were checked in the villages without water-improvement,incidence of endemic arsenicosis was 3.08％ (189/6166).Among them,mild patients was 2.69％ (166/6166),moderate was 0.28％ (17/6166) and the serious was 0.10％ (6/6166); 20 new cases were diagnosed,and they came from Shanxi province.Conclusions The morbidity in water-improved villages remains higher than the water-unimproved.We should establish and perfect the long-term mechanism of surveillance as soon as possible,and strengthen the management and maintenance of water-improving projects.

Effects of edaravone on the expression of GRP78, Caspase-12, and neuron apoptosis in juvenile rat hippocampus after status convulsive / 中华儿科杂志

<p><b>OBJECTIVE</b>To observe the expression of GRP78 (glucose regulated protein, GRP78), Caspase-12 and the change of neuron apoptosis in the juvenile rat hippocampus after status convulsive (SC), and to explore the effect of edaravone on them.</p><p><b>METHODS</b>One hundred and ninety-five juvenile male Sprague-Dawley (SD) rats were randomly divided into normal saline control group (NS group), status convulsive group (SC group) and edaravone treatment group (ED group). Each group was further divided into five subgroups in different executed time points after SC. The rats in status convulsive group were kindled into epilepsy by lithium-pilocarpine method. Expression of GRP78 mRNA and caspase-12 mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR) method. Expressions of GRP78 and caspase-12 protein were detected with immunohistochemical methods. The neuron apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL).</p><p><b>RESULTS</b>(1) Measured by immunohistochemistry the value of OD of GRP78 (0.1480 ± 0.0164, 0.1682 ± 0.0114, and 0.1540 ± 0.0102, respectively, 12 h - 48 h points) and caspase-12 (0.1325 ± 0.0165, 0.1794 ± 0.0213, 0.1525 ± 0.0423, and 0.1309 ± 0.0199, respectively, 12 h-72 h points) positive cells in the SC group increased, there was a significant difference compared with NS group (GRP78: 0.1214 ± 0.0147, 0.1272 ± 0.0177, and 0.1260 ± 0.0157, respectively, 12 h-72 h points. Caspase-12: 0.1050 ± 0.0121, 0.1041 ± 0.0151, 0.1058 ± 0.0222, and 0.1036 ± 0.0186, respectively, 12 h - 72 h points) (P < 0.01, or P < 0.05). By ED intervention GRP78 (0.1550 ± 0.0131, 0.1886 ± 0.0154, and 0.1721 ± 0.0151, respectively, 12 h - 48 h points) positive cells value of the OD increased as compared with SC group (P < 0.01, or P < 0.05). and caspase-12 (0.1211 ± 0.0184, 0.1545 ± 0.0205, and 0.1085 ± 0.0219, respectively, 12 h, 24 h and 72 h points) positive cells value of the A decreased as compared with SC group (P < 0.01, or P < 0.05). (2) Measured by RT-PCR, the expression of GRP78 mRNA and caspase-12 mRNA trend was similar to protein. (3) The TUNEL positive cells in hippocampus CA(1) of SC group (11.41 ± 2.37) were more than that of NS group after the SC 12 h (P < 0.01), reached its highest level at 48 h (28.78 ± 5.11), after the intervention with edaravone (8.98 ± 2.22, 13.09 ± 2.54 and 20.57 ± 4.89, respectively, 12 h-48 h points), TUNEL positive cells showed a significant drop in SC group at 12 h-48 h time points (P < 0.01, or P < 0.05), but still significantly higher than that of the NS group (6.22 ± 1.50, 6.57 ± 1.61 and 6.72 ± 1.14, respectively) (P < 0.01, or P < 0.05), at the 4 h time point (NS group 6.29 ± 1.49, SC group 6.61 ± 1.71, ED group 5.75 ± 1.41) among the three groups, no significant difference in TUNEL positive cells was found (P = 0.759).</p><p><b>CONCLUSIONS</b>The expression of GRP78 and caspase-12 increased after SC. Edaravone increased expression of GRP78 and decreased expression of caspase-12 in hippocampus rat with pilocarpine-induced seizures, reduced the number of neuronal apoptosis. These results suggest that edaravone may have protective effect against the hippocampal damage caused by status convulsive.</p>

Effects of W-7 on the expression of GRP78 and neuronal apoptosis in immature rat hippocampus after status convulsion / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effects of the calmodulin inhibitor W-7 on the expression of the key marker of ERS GRP78 and neuronal apoptosis in the immature rat hippocampus after status convulsion (SC).</p><p><b>METHODS</b>One hundred and seventeen male Sprague-Dawley rats aged 19-21 days were randomly divided into three groups: normal saline control (control), SC with and without W-7 pretreatment. Each of the 3 groups was further subdivided into subgroups sacrificed at 4, 24 and 48 hrs. SC model was prepared using lithium-pilocarpine. GRP78 mRNA expression in the hippocampus was detected by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). GRP78 protein was ascertained by immunohistochemistry. Neuronal apoptosis was observed with TdT-mediated dUTP nick end labeling (TUNEL).</p><p><b>RESULTS</b>The expression of GRP78 mRNA was significantly increased in the non-pretreated SC group compared with the control group 24 hrs after injection of saline or lithium-pilocarpine (P<0.01), and the expression of GRP78 protein also increased markedly in the seizure group compared with the control group 24 and 48 hrs after the injection (P<0.01). The expression of GRP78 mRNA and protein in the W-7 pretreatment group was significantly higher than both the control and the non-pretreated seizure groups 24 and 48 hrs after injection. The TUNEL positive cells in the hippocampus CA1 in the non-pretreated SC group 24 and 48 hrs after injection (21.0 ± 2.5 and 29.4 ± 2.8, respectively) were increased compared to the control group (7.1 ± 1.4 and 7.3 ± 1.6, respectively; P<0.01). W-7 pretreatment decreased TUNEL positive cells to 15.0 ± 2.5 and 20.0 ± 2.9 at 24 and 48 hrs after injection compared to the non-pretreated seizure group (P<0.01), but the number of TUNEL positive cells in the W-7 pretreatment group remained significantly greater than in the control group (P<0.01).</p><p><b>CONCLUSIONS</b>W-7 may up-regulate the expression of GRP78 and reduce the number of apoptotic neurons, thus provides a neuroprotective effect against brain damage following SC.</p>

Effects of edaravone on glial fibrillary acidic protein and interleukin-lβ expression and neuronal apoptosis in juvenile rat hippocampus after status convulsion / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of edaravone on glial fibrillary acidic protein (GFAP) and interleukin-1β (IL-lβ) expression and neuronal apoptosis in the juvenile rat hippocampus after status convulsion (SC).</p><p><b>METHODS</b>One hundred and ninety-five juvenile male Sprague-Dawley (SD) rats were randomly divided into 3 groups: normal saline control and SC with and without edaravone treatment. Each of the 3 groups was further subdivided into subgroups sacrificed at 4, 12, 24, 48 and 72 hrs after SC (n=15). The SC model was prepared using lithium-pilocarpine. The expression of GFAP and IL-lβ protein was detected with immunohistochemistry methods. The neuronal apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL). The hippocampal GFAP mRNA expression was detected by RT-PCR.</p><p><b>RESULTS</b>The value of IOD of GFAP and IL-lβ positive cells measured by immunohistochemistry in the untreated SC group increased compared with the control group. Expression of GFAP and IL-lβ protein was significantly reduced in the edaravone treated SC group compared with the untreated SC group. RT-PCR showed the expression trend of GFAP mRNA was similar to that of protein. The TUNEL positive cells in the hippocampus CA1 in the untreated SC group increased significantly 12 hrs after SC and reached a peak at 48 hrs compared with the control group. The intervention with edaravone decreased significantly TUNEL positive cells between 12-48 hrs after SC, but the number of TUNEL positive cells in the intervention group remained significantly greater than in the control group.</p><p><b>CONCLUSIONS</b>The expression of GFAP and IL-lβ in the hippocampus increases after SC in rats. Edaravone may decrease the expression of GFAP and IL-1β and reduce the number of neuronal apoptosis. These results suggest that edaravone may have protective effects against brain damage caused by SC.</p>

Expression of brain-derived neurotrophic factor and apoptosis in the brain of rats with repeated febrile seizures / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes of brain-derived neurotrophic factor (BDNF) following repeated febrile seizures in rats and its possible correlation with neurocyte apoptosis.</p><p><b>METHODS</b>Fifty-one male Sprague-Dawley (SD) rats were randomly assigned to three groups: normal control (n=14), febrile seizure (FS, n=18), hyperthermia alone (n=19). Febrile seizures were induced by hot water bath. The level of BDNF in the hippocampal homogenate was measured using ELISA and the expression of BDNF in various brain regions was measured by immunohistochemistry. The neurocyte apoptosis of the brain was determined by TdT-mediated biotinylated-dUTP nick end labling (TUNEL).</p><p><b>RESULTS</b>The level of BDNF in the hippocampus in the FS group(89.9+/-12.5 ng/g)was higher than that in the normal control group(54.4+/-18.9 ng/g)and in the hyperthermia alone group (64.1+/-15.0 ng/g) (P<0.01). The OD value of BDNF positive neurons in various brain regions of the FS group was significantly higher than that of the normal control group (P<0.01) and the hyperthermia alone group (P<0.01). The FS group had significantly higher apoptotic index in various brain regions than the normal control and the hyperthermia alone groups (P<0.01). There was a positive correlation between the expression of BDNF and the apoptotic index in various brain regions (r=0.332, P<0.05).</p><p><b>CONCLUSIONS</b>BDNF expression in the brain increases following repeated febrile seizures in rats, and the increased BDNF expression is correlated with neurocyte apoptosis.</p>

Protection of edaravone on neurons and its effects on the expression of interleukin-lbeta in juvenile rat hippocampus following status convulsion / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the possible protection of edaravone on neurons of the hippocampus after status convulsion (SC) and its effects on the expression of interleukin-1beta (IL-lbeta) in juvenile rats.</p><p><b>METHODS</b>One hundred and ninety-five juvenile male Sprague-Dawley rats were randomly divided into three groups: SC, edaravone pretreatment and normal saline control (control group). Each group was subdivided into five groups sacrificed at 4, 12, 24, 48 and 72 hrs after SC induction. SC model was prepared using lithium-pilocarpine. The edaravone pretreatment group received edaravone by intraperitoneal injection once daily three days before convulsion induction. Histopathologic changes in the hippocampus were viewed under a light microscope and an electron microscope. Expression of apoptosis cells was observed by TdT-mediated dUTP nick end labeling (TUNEL). Expression of IL-lbeta protein was determined by immunohistochemistry.</p><p><b>RESULTS</b>Under the electron microscrope, a small quantity of neurons showed karyopycnosis and endocytoplasmic reticulum (ER) expanded remarkably 24 hrs after SC induction; at 48 hrs the ER expanding was alleviated somewhat but mitochomdria swelling was more severe. The edaravone pretreatment group showed less severe neuronal changes compared with the SC group under the microscopes. The TUNEL positive cells in the hippocampus of the SC group were significantly more than those of the control group 12 hrs, and peaked at 48 hrs after SC induction. The edaravone pretreatment group showed decreased TUNEL positive cells in the hippocampus compared with the SC group, although the positive cells were more than those in the control group between 12 and 48 hrs after SC induction. The immunohistochemistry assay demonstrated that the expression of IL-lbeta in the hippocampus of the SC group increased significantly compared with that of the control group 12, 24, 48 and 72 hrs after SC induction. Edaravone pretreatment resulted in a significantly decreased IL-lbeta expression in the hippocampus as compared with the SC group.</p><p><b>CONCLUSIONS</b>Edaravone pretreatment may decrease the IL-1beta expression and neuronal apoptosis in the hippocampus. This suggests that edaravone may have protective effects against the hippocampal damage caused by SC.</p>

Clone screening and interference efficiency of seed cells with CCL20 gene knockdown for tissue-engineered skin / 中华外科杂志

<p><b>OBJECTIVE</b>To screen stable cell clones of CCL20 gene knockdown and assess their interference effects, recombinant lentivirus vectors with CCL20 gene specific shRNA were applied to infect human immortal keratinocyte line (HaCaT).</p><p><b>METHODS</b>The three pHSER-CCL20-shRNA-GFP vectors (pHCG-1 and pHCG-2 were CCL20 gene specific, and pHCG-3 was used as mismatch control) have been previously constructed. The virus packaging cell line 293FT was transfected with these vectors by using CaCl2 methods to produce lentiviral particles. After the viral titers of these three harvested cell supernatants were determined by flow cytometry, HaCaT cells were transfected by these viruses and screened under the pressure of G418. The CCL20 mRNA from HaCaT cell clones and the CCL20 protein levels in the supernatants of HaCaT cell clones were detected by Real-time RT-PCR and ELISA, respectively.</p><p><b>RESULTS</b>The titers of three lentiviruses were 7.08 x 10(5) transduced units (TU)/ml, 1.88 x 10(5) TU /ml and 2.08 x 10(5) TU/ml, respectively. Two HaCaT cell clones from each lentiviral vectors were obtained after G418 screening for 5 - 8 weeks. Four CCL20 gene specific clones showed stable interference effect in both Real-time RT-PCR and ELISA. The mRNA expression and protein level of CCL20 gene specific clones were down regulated significantly.</p><p><b>CONCLUSIONS</b>The four human immortal keratinocyte clones with long term CCL20 gene knockdown have been screened by recombinant lentivirus vectors with CCL20 gene specific shRNA. These clones might be served as seed cells for novel tissue-engineered skin with lower rejection.</p>

Single nucleotide polymorphisms and haplotypes analysis of DFNB1 locus in Chinese sporadic hearing impairment population / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The DFNB1 locus, which contains the gap junction beta-2 (GJB2) and gap junction beta-6 (GJB6) genes, plays a key role in the nonsyndromic and sporadic hearing impairment. Mutations of DFNB1 result in autosomal recessive nonsyndromic hearing impairment (ARNSHI). Previous researches have identified mutations in GJB2 and GJB6, but single nucleotide polymorphisms (SNPs) of DFNB1 locus have not been studied. So we chose five SNPs to evaluate whether there is difference between deafness people and normal-hearing people in Han Chinese.</p><p><b>METHODS</b>Five SNPs in the DFNB1 region were examined using a case-control association study between cases with sporadic hearing impairment and controls with normal hearing. The HWEsoft and SHEsis softwares were used to analyze the results.</p><p><b>RESULTS</b>Single-locus association analysis showed a positive association for three SNPs: rs9315400, rs2274084 and 235delC. When we compared the distributions of the haplotypes, we also found significant differences between cases and controls in the haplotype combination of rs2274084 and rs2274083 (chi(2) = 12.978, df = 3, global P = 0.004719).</p><p><b>CONCLUSIONS</b>The haplotypes composed of rs2274084 and rs2274083 suggested that C-C may be a risk haplotype for the sporadic hearing impairment while T-T may be protective against hearing impairment. From that point of view, we can conclude that the SNPs of DFNB1 locus also plays an important role in sporadic hearing impairment cases.</p>

Effects of edaravone on IRE1 mRNA expression and neuronal apoptosis in the hippocampus of rats with status convulsivus / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the expression of the key marker of endoplasmic reticulum stress (ERS) IRE1 mRNA and neuronal apoptosis in the rat hippocampus after status convulsivus (SC), and the intervention effects of edaravone, a novel free radical scavenger.</p><p><b>METHODS</b>Sprague-Dawley (SD) rats aged 19-21 days were randomly assigned to three groups: normal control, SC and edaravone-treated SC. SC was induced in the later two groups. The two groups were subdivided into 5 groups sacrificed at 4, 12, 24, 48, and 72 hrs after SC induction. IRE1 mRNA expression in the hippocampus was detected by RT-PCR. Neuronal apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL). The ultramicrostructural changes of neuron were observed by electron microscopy.</p><p><b>RESULTS</b>IRE1 mRNA expression was obviously up-regulated 4 and 12 hrs after SC compared with the normal control group (P<0.01). IRE1 mRNA expression in the edaravone-treated SC group was notably higher than the untreated SC group 4, 12 and 24 hrs after SC and the normal control group (P<0.01). TUNEL positive cells in the hippocampus in the untreated SC group were significantly more than those in the normal control group (P<0.01). The number of TUNEL positive cells increased with the prolonged convulsion time. TUNEL positive cells in the edaravone-treated SC group were significantly reduced compared with those in the untreated SC group 12, 24, 48 and 72 hrs after SC (P<0.05 or P<0.01), but remained higher than the normal control group (P<0.05 or P<0.01). The peri-nucleus cell organ injuries were observed 4 hrs after SC and karyopycnosis and cytoplasm condensation were observed 12 hrs after SC in the SC and the edaravone-treated SC groups. The edaravone-treated SC group demonstrated less severe apoptosis than the untreated SC group.</p><p><b>CONCLUSIONS</b>Edaravone may have neuroprotections against SC by an up-regulation of IRE1 expression. It might serve as an effective agent for reducing ERS in vivo.</p>