Objective: To observe the improving effect of muscle regions of meridians needling method on the upper limb function in children with cerebral palsy of spastic hemiplegic type. Methods: A total of 100 children with cerebral palsy of spastic hemiplegia type were divided into a treatment group and a control group according to the visiting sequence, with 50 cases in each group. The control group was treated with conventional rehabilitation plus conventional acupuncture treatment. The treatment group was treated with conventional rehabilitation plus muscle regions of meridians needling method. The electromyography (EMG) signal values of triceps brachii and pronator teres were detected before treatment, and 3 months and 6 months after treatment. The clinical efficacy was evaluated by Peabody developmental motor scale-fine motor (PDMS-FM) and fine motor function measure (FMFM). Results: Three and six months after treatment, the EMG signal values of triceps brachii and pronator teres, grasping scores and visual-motor integrated scores of PDMS-FM and the FMFM scores in both groups increased to varying degrees compared with the same group before treatment, and the intra-group differences were all statistically significant (all P<0.05). Six months after treatment, the results of the above three items in the treatment group were all better than those in the control group, and the differences between the groups were statistically significant (all P<0.05). Conclusion: Muscle regions of meridians needling method added on the basis of conventional rehabilitation can effectively reduce the muscle tone of upper limb and enhance the muscle strength, and improve the upper limb function in children with cerebral palsy of spastic hemiplegia type. The efficacy is superior to that of the conventional rehabilitation plus conventional acupuncture treatment.

Objective To investigate the effects of sufentanil on activation of spinal astrocytes in mice after unilateral sciatic nerve injury.Methods Eighty healthy male BALB/c mice,aged 6-8 weeks,weighing 18-22 g,were randomly divided into 4 groups (n=20 each): sufentanil high dose group (group H),middle dose group (group M),low dose group (group L) and model group (group MO).After model of sciatic nerve injury was established by unilateral sciatic nerve transection,group H,M and L,sufentanil 10 μg/kg,5 μg/kg,2.5 μg/kg was injected intraperitoneally once a day continuously for 3 days,while the equal volume of saline was injected in group MO.Five mice in each group were selected,and the sciatic nerve functional index were measured at 4,8,12 weeks.The 5 mice were sacrificed from each group and the damage on the same side L4-L6 segments of the spinal cord were removed at 2,4,8,12 weeks.The pathological changes were examined under light microscope at 8 week point.The expression of GFAP was determinatied at each time points by immuno-histochemistry.Results Compared with group L and MO,sciatic nerve functional index significantly was increased in groups H and M (P<0.05),and no significantly change was found in group L.Spinal cord neurons had a better morphology in groups H and M than in group L and MO.Compared with group MO,The expression of GFAP were significantly up-regulated in groups H,M and L (P<0.05).Compared with group L,the expression of GFAP were significantly up-regulated in groups H and M (P<0.05).Conclusion Sufentanil promotes spinal astrocyte activation after peripheral nerve injury in mice and improves repair after peripheral nerve injury.

Objective To evaluate the effects of sufentanil on regeneration and repair after single sciatic nerve injury in mice. Methods Seventy?five healthy male BALB∕c mice, aged 6-8 weeks, weig?hing 18-22 g, were divided into 5 groups ( n=15 each) using a random number table: peripherial nerve injury group (group PNI), low, medium and high doses of sufentanil groups (L, M and H groups) and cyclosporine A group ( group C) . The model of unilateral sciatic nerve transection was established in the 5 groups. In L, M and H groups, sufentanil 2?5, 5?0 and 10?0 μg∕kg were injected intraperitoneally, re?spectively, once a day for 3 consecutive days. Cyclosporine A 50 mg∕kg was injected intraperitoneally once a day for 3 consecutive days in group C. The equal volume of normal saline was given once a day for 3 con?secutive days in group PNI. Neurophysiological monitoring was performed at 4, 8 and 12 weeks after opera?tion, the amplitude of compound muscle action potentials of the sciatic nerve was recorded, and the nerve conduction velocity was measured. At 8 weeks after operation, 5 mice were sacrificed, the sciatic nerve 0?5 cm of the upper and lower the anastomosed site was removed for examination of the morphology of myelin sheath with light microscope, and the number of nerve fibers was calculated. Results Compared with group PNI, the amplitude of compound muscle action potentials of the sciatic nerve, nerve conduction ve?locity and the number of nerve fibers was were significantly increased in M, H and C groups ( P<0?05) , and no significant change was found in the parameters mentioned above in group L ( P>0?05 ) . Myelin sheath arrangement was severely irregular, and more vacuoles were found in group PNI. Myelin sheath ar?rangement was irregular, and more vacuoles were found in group L. Myelin sheath arrangement was mainly regular, and vacuoles were found occasionally in group M. Myelin sheath arrangement was regular, and no vacuoles were observed in H and C groups. Conclusion Sufentanil can promote regeneration and repair af?ter peripheral nerve injury in mice.

Objective To study the expression of thymic stromal lymphopoietin(TSLP) and the activation of DCs in OVA-induced murine asthma model, and investigate the effects and underlying mecha-nisms of TSLP on lung inflammation. Methods Thirty BALB/c mice were randomly divided into control group, OVA group and TSLP neutralizing antibody treated group. The asthma model was evaluated by airway responsiveness and histological analysis of lung tissues ; The levels of TSLP mRNA in lungs were determined by quantitative real-time PCR; The expression of TSLP in lungs were determined by immunohistochemistry and Western blot; The expression of CD40, CD80, CD86 in BALF was detected by FACS. Results Both the histological analysis of lung tissues and the airway responsiveness were all consistent with the characteris-tic of murine asthma model. The expression of TSLP and TSLP mRNA in the OVA group was significantly in-creased compared with blank group. The expression of CD40, CD80, CD86 in BALF from OVA group was increased significantly compared with the control group. Furthermore, treating mice with TSLP neutralizing antibody reduced the expression of CD40, CD80, CD86 on dendritic cells, and IL-4, IL-5, IL-13 in the OVA group. Conclusion Our study indicate that TSLP was highly expressed in the bronchial epithelia of murine asthma model, via upregulation of CD40, CD80, CD86, induce DCs to active CD4~+ T cells and pro-duce type 2 responses, so that aggravating the lung inflammation of asthma. Blocking TSLP is capable of in-hibiting the production of Th2 cytokines, thus presents a promising strategy for the treatment of asthma.

Objective To observe the effect of inducible costimulator(ICOS) costimulation pathway blockade in rat limb allografts acute rejection by RNA interference. Methods Twenty-seven cases of modified hind llmb allotransplantation were performed from Wistar to SD rats. The rats were divided into 3 gronps(each n=9): the rejection group not given a special disposal; the control group, consisting of SD rats that received injection of pSilencer 4.1 and Sofast complex by vein post transplantation; and the interference group that received injection of pSilencer 4.1-ICOSshRNA and Sofast complex. On the eighth day posttransplantation, 3 rats were killed to study the pathological changes in each group. The expressions of ICOS gene in vivo were detected by flow cytometry and RT-PCR. The mixed lymphocyte reaction (MLR) was performed and eytokines in blood were measured by ELISA. The rest rats were used to record limb survival time. Results The mean survival time in rats of the rejection and the control groups were(11.34±1.21) and (11.14±1.32) days respectively. In the interference group, the mean survival time of limb allografts was (16.85±1.73) days(P<0.05). The rats in the rejection and the control groups experienced moderate to serious acute rejections with skin epidermal necrosis, a large quantity of lymphocyte infdtration, muscle cell necrosis and interstitial edema, while the pathological changes in rats of the interference group were mild. The splenocyte ICOS mRNA expression level in the interference group(18.75%) was significantly lower than that of the rejection group(100%) and the control group(98.51%). ICOS cell surface expression level as judged by the fluorescence intensity was 45.59±12.87 in the interference group, 103.72±21.76 in the rejection group, and 93.47±29.55 in the control group(F=6.89, P<0.05). In stimulation assays, a one-way mixed lymphocyte reaction stimulation index(SI), with spleen cells from Wistar and Lewis rats, respectively, the rejection group (5.26±0.42,5.18±0.29) and the control group (5.37±0.27,4.93±0.44) had significantly greater reactions than the interference group(2.37±0.35, 4.87±0.36), respectivily(F=7.29, P<0.05; F=6.19, P0.05). In the IFN-γ and IL-4 expression assays, reactions of the interference group (230.17±38.47,160.32±59.13) were lower than those of the rejection group(490.73±51.48,230.67±45.21) and the control group(480.15±43.96, 240.53± 47.36), (F=7.23,6.75, all P<0.01). Conclusions In vivo transfection of pSilencer 4.1-ICOS shRNA interference plasmid can effectively block T-cell co-stimulation pathway, suppress acute rejection, and prolong limb allografts survival.

Objective:To observe the clinical efficacy of needling thirteen ghost acupoints for children with autism spectrum disorder. Methods:A total of 90 cases with autism spectrum disorder (ASD) aged between 2 and 6 years were randomly allocated into 2 groups by random number table. The control group (n=45) received routine rehabilitative training, and the treatment group (n=45) received acupuncture at thirteen ghost acupoints plus routine rehabilitative training. The Beijing Gesell developmental (Gesell) scale and autism behavior checklist (ABC) were used to assess the intellectual, language and behavior development before and 3 months after the treatment. Results:After the treatment, the total effective rate in the treatment group was 82.2%, versus 55.6% in the control group, showing a statistical significance (P<0.05). As for the scores of social, emotional and language in Gesell scale, there were significant intra-group differences in the treatment group (allP<0.05), and all the five subscales in the Gesell scale in the treatment group were significantly better than those in the control group (allP<0.05). As for the scores of ABC, there were significant intra-group differences in the treatment group (P<0.05), and the scores in the treatment group were significantly better than those in the control group (allP<0.05). Conclusion: Rehabilitation training plus acupuncture at thirteen ghost acupoints can significantly improve the intellectual, language and abnormal behavior in autism spectrum disorder children.

OBJECTIVETo optimize formulation of tanshinone II(A)-loaded PLGA nanoparticles and compare the difference of two methods in preparation and quality of nanoparticles.

METHODThe two methods were nanoprecipitation method and emulsion-evaporation method. Single factor experiments and central composite design and response surface method were used to optimize the formulation of nanoparticles. The nanoparticles were characterized at size, morphology, entrapment efficiency, drug loading, drug recovery rate, crystallinity and drug release in vitro.

RESULTThe mean diameters were 225 nm and 183 nm, the entrapment efficiency were 95.49% and 87.99%, the drug loading were 2.03% and 0.16%, and the drug recovery rates were 38.42% and 17.59% respectively for nanoprecipitation method and emulsion-evaporation method.

CONCLUSIONNanoprecipitation method was better than emulsion-evaporation method for preparation of tanshinone II(A)-loaded PLGA nanoparticles.

Chemical Precipitation ; Crystallization ; Diterpenes, Abietane ; Emulsions ; Lactic Acid ; chemistry ; Nanoparticles ; chemistry ; Particle Size ; Phenanthrenes ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polyglycolic Acid ; chemistry ; Polymers ; chemistry ; Quality Control ; Salvia miltiorrhiza ; chemistry ; Technology, Pharmaceutical ; methods ; Volatilization

Objective To explore the effect of Jian Zu San Zhen acupuncture on tip foot in children with spastic cerebral palsy. Methods From January to December, 2016, 120 children with spastic cerebral palsy aged one to six years were randomly divided into control group (n=60) and observation group (n=60). Both groups were treated with conventional rehabilitation, while the observation group added Jian Zu San Zhen acupuncture. They were evaluated with Gross Motor Function Measure (GMFM), foot dorsiflexion angle and surface electromyog-raphy. Results After treatment, the scores of GMFM in B, C, D and E dimensions and the total score significantly increased in the observa-tion group (t>4.325, P<0.001), and were higher than that of the control group (t>2.711, P<0.01);the foot dorsiflexion angle significantly de-creased in the observation group (t=17.627, P<0.001), and was significantly less than that of the control group (t=15.416, P<0.001). The root mean square (RMS) of surface electromyography of gastrocnemius muscle under active plantarflexion and passive dorsiflexion of ankle joint improved after treatment (t≥3.058, P<0.01), and was better in the observation group than in the control group (t=-2.199, P<0.05). Conclusion Jian Zu San Zhen acupuncture can effectively reduce the gastrocnemius muscle tension, and do not reduce the gastrocnemius muscle strength, which could also improve the ability of foot flexion, and facilitate to recover the function of standing and moving.

A rapid UPLC method for simultaneous determination of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus was established. The optimal conditions of separation and detection were achieved on an HSS T3 Column (2.1 mm x 100 mm, 1.8 microm) eluted with a gradient of acetonitrile and 0.05% aqueous phosphoric acid, at a flow rate of 0.5 mL x min(-1), detected at 254 nm. The linear response ranges of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde were 0.359-3.59 mg x L(-1) (r = 0. 999 3), 2. 834-28.34 mg x L(-1) (r = 0.999 8), 0.574-5.74 mg x L(-1) (r = 0.999 8), 2.400-24.00 mg x L(-1) (r = 0.999 9), 32.57-325.7 mg x L(-1) (r = 0.999 8), respectively (n = 6). The mean recoveries (n = 9) of the five components were 96.7%-101.0%, RSD < 2.3%. The assay demonstrates that the method has adequate accuracy and selectivity to measure the concentration of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus.
Acrolein ; analogs & derivatives ; analysis ; Chromatography, Liquid ; methods ; Cinnamates ; analysis ; Cinnamomum ; chemistry ; Coumarins ; analysis ; Drugs, Chinese Herbal ; chemistry ; Hydroxybenzoates ; analysis ; Propanols ; analysis

AIMTo study the preparation of bovine serum albumin nanoparticles surface-modified with glycyrrhizin(BSA-NP-GL) targeting to hepatocytes.

METHODSThe bovine serum albumin nanoparticles (BSA-NP) were prepared by desolvation process. Glycyrrhizin (GL) was oxidized by sodium periodate to be conjugated to surface reactive amino groups (SRAG) of the BSA-NP. The SRAG were quantified by spectrophotometric method using 2, 4, 6-trinitrobenzenesulfonic acid(TNBS). Glycyrrhetinic acid(GA) hydrolyzed from GL, which was on the surface of BSA-NP-GL was assayed by HPLC after isolation by sephadex G-50. Both methods were used to verify the conjugation achieved. HPLC was used to determine surface density of GL on BSA-NP-GL.

RESULTSThe amount of SRAG of the BSA-NP-GL decreased by 19.6% compared with normal BSA-NP. The amount of GL molecule was 9.2% of the total determined SRAG of BSA-NP. The mean diameter of the BSA-NP-GL was 73 nm with round shape. The stability of BSA-NP-GL was constant when it was stored at 25 degrees C and 37 degrees C during 10 days.

CONCLUSIONBSA-NP-GL was successfully prepared, which is considered to establish an experimental foundation for further research on its ability for targeting to hepatocytes.

Cross-Linking Reagents ; chemistry ; Drug Delivery Systems ; Glycyrrhizic Acid ; chemistry ; Nanotechnology ; Particle Size ; Serum Albumin, Bovine ; administration & dosage ; chemistry ; ultrastructure ; Surface Properties ; Technology, Pharmaceutical ; methods