Objective To investigate the relationship between cleftpalate and secretory otitis media so that the secretory otitis media resulted from cleftpalate was treated and prevented effectively. Methods 116 children with cleftpalate and 73 normal children were investigated by tympanogram, hearing threshold and auditory brain response. Results The incidence of secretory otitis media and hearing loss had significant difference between the two groups. Conclusions Children with cleftpalate had high incidence of secretory otitis media and hearing loss. We should pay more attention to the treatment and prevention of secretory otitis media in children with cleftpalate. It is necessary for the cleftpalate children to insert the tympanal pressure equalization tubes while palatal surgery is performed.

Objeetlve To evaluate the role of IL-1β and TNF-α in the spinal cord in a murine model of bone cancer pain(BCP)induced by activation of glial cells.Methods Three hundred and sixty male 8-10 weeks old C3H/He mice weighing 18-22 g were randomly divided into 6 groups(n=60 each):group I nomal control (group N);group Ⅱ sham operation (group S);group Ⅲ BCP+aCSF(group aCSF);group Ⅳ BCP+FC (group FC);group V BCP+MI(group MI) and group Ⅵ BCP+FC+MI (group FC-MI).BCP was produced by inieeting fibrosarcoma cells of bone into the medullary cavity of left ealcaneus bone.Intrathecal catheter was placed in the 4 BCP groups(group Ⅲ-Ⅳ).FC 0.5 nmol/5 μl or/and MI 16 μg/5 μl were injected IT once a day for 21 consecutive days after operation.The mechanical threshold to von Frey filaments was measured at 0.5 h(T0)before injection of fibrosarcoma cells and at 3,5,7,10,14,21 d(T1-6)after injection of fibrosarcoma cells.Twelve animals of each group were killed and L4.5 segment of the spinal cord was removed at T0,1,3,5,6 for determination of IL-1β and TNF-α content (by ELISA) and expression (by immuno-flurorescence) in the spinal cord. Results The mechanical threshold was significantly decreased at T1-6, while IL-1β content at T1,3,5,6 and TNF-α content at T5,6 was significantly increased in group BCP, FC, MI and FC + MI compared with those at T0 and group C (P < 0.05). Compared with group BCP, the mechanical threshold was significantly increased at T1-6 in group MI and FC + MI and at T4-6 in group FC, IL-1β content was significantly decreased at Ts,3,5,6 in group MI and FC + MI and at T5,6 in group FC and TNF-α content was significantly decreased at T5,6 in group FC, MI and FC + MI ( P < 0.05). Conclusion IL-1β and TNF-α in the spinal cord is involved in the process of glial cell activation-induced BCP.

ObjectiveTo investigate the effects of Akt3 gene knockout on neuropathic pain behaviors induced by chronic constriction injury of sciatic nerve (CCI).MethodsExperiment was divided into two groups:Akt3 knockout group (Akt3-/-,n =12),wild type group (WT,n =12 ).Randomly numbered,the right sciatic nerve of mice were received the operation of chronic constriction injury.Paw withdrawal mechanical threshold (PWMT)and paw withdrawal thermal latency (PWTL) were tested on day 1 before operation and day 1,3,5,7,10,14,17,21 afterCCI.ResultsThe basic values of PWMT(right:(1.09±0.20)g,(1.17±0.22)g;left:(1.17±0.15)g,(1.22±0.23)g,P＞0.05) andPWTL(right:(6.18±1.11)s,(6.20±1.25)s;left:(5.82±0.91)s,(5.92± 1.71 ) s,P ＞ 0.05 ) had no statistically significant differences between two groups.On day 1 after operation,compared with basic values,the PWMT and PWTL of the right paw in both Akt3-/- group and WT group decreased significantly (P ＜ 0.05 ),and at least lasted up to day 21.The PWMT( 3d:(0.42 ± 0.22 ) g,(0.72 ± 0.36) g ; 17d:(0.29 ±0.15)g,(0.49 ±0.19) g;21d:(0.27 ±0.18)g,(0.56 ±0.15)g,P＜0.05) and PWTL(5d:(2.43 ±0.68)s,(3.13±0.52)s;17d:(2.43±1.26)s,(3.84±1.29)s ;21d:(2.14±1.23)s,(4.07±1.26)s,P＜0.05 ) of the right paw in Akt3-/- group was significantly lower than those in WT group.The PWMT and PWTL of the left paw in Akt3-/- group and WT group had no obvious differences (P ＞ 0.05 ). However.compared to left paw,the PWMT and PWTL of the right paw of the two groups were obviously lower (P ＜ 0.05 ).ConclusionThe neuropathic pain induced by CCI increased in Akt3 gene knockout mice.

Objective To investigate the effects of different doses of intrathecal magnesium on bone cancer pain (BCP) in mice.Methods Two hundred and eighty-eight male C3H/HeJ mice,aged 8-10 weeks,weighing 18-22 g,were randomly divided into 6 groups (n =48 each):control group (group C) ; sham operation group (group S) ; BCP + artificial cerebro-spinal fluid (aCSF) 5 μl group (group BCP) ; BCP + MgSO4 14.4 μg group (group M1 ) ; BCP + MgSO4 43.2 μg group (group M2 ) and BCP + MgSO4 86.4 μg group (group M3 ).BCP was produced by injecting fibrosarcoma cells of bone into the medullary cavity of right femur.Intrathecal catheter was placed in the 4 BCP groups.The aCSF 5 μl or MgSO4 14.4μg/5 μl,43.2 μg/5 μl,or 86.4μg/5 μl was injected intrathecally on 14th day after inoculation of tumor cells.The paw withdrawal threshold to mechanical stimuli (PWMT) and paw withdrawal lateney to thermal stimuli (PWTL) were measured at 0.5 h before administration (T0 ) and at 0.5,2,4 and 8 h after administration (T1-4).Eight animals chosen from each group at T0-4 were sacrificed,and L4-5 segment of the spinal cord was removed for determination of NR2B expression (by immuno-flurorescence) in the spinal cord.Results PWMT and PWTL were significantly decreased at T0-4,and NR2B expression was significantly up-regulated at T0-4 in groups BCP,M1,M2,M3 compared with groups C and S ( P ＜0.05).Compared with group BCP,PWMT and PWTL were significandy increased at T1-3,and NR2B expression was significantly down-regulated at T1-3 in groups M2 and M3 ( P ＜ 0.05).Compared with group M2,PWMT and PWTL were significantly increased at T1-3,and NR2B expression was significantly down-regulated at T1-3 in group M3 ( P ＜ 0.05).Conclusion Intrathecal magnesium can reduce BCP in a dose-dependent manner in mice.

Compound Danshen Dripping Pills has curative effect of angina pectoris,coronary heart disease,and other cardiovascular and cerebrovascular diseases,which is widely used in clinic.With the development of basic research and clinical pharmacology,it is particularly important to carry out more comprehensive and systematic quality control.At present,fingerprint technology is developing rapidly and has been recognized by many countries.In this article,we summarized the research situation of Compound Danshen Dropping Pills and its raw materials from two aspects of chromatography and spectroscopy (such as LC,CE,IR,NMR and so on).It will provide reference or the further establishment of a reasonable quality control method.

Objective To provide a proper concentration of Internal Standard Solution by comparing five different degrees to the simultaneous analysis of 10 metals including V,Cr,Mn,Fe,Ni,Cu,As,Cd,Pb and Hg in Cardeniajasminoides.Method An Inductively Coupled Plasma-Mass Spectrometry method with Microwave-Assisted Digestion was developed with10,50,200,500,1 000 μg/L of Ge,In,Bi as internal standard.Result The degree of 500 μg/L concentration of internal stand solution can both correct the matrix effect and alleviate the contamination as the 1 000 μg/L.Conclusion With excellent detection limits,high sensitivity and wide linear range,the 500 μg/L internal standard may serve as reference in the determination of elements in other traditional Chinese medicine (TCM).

OBJECTIVE:To establish a method for simultaneous determination of 18 toxic elements from 10 ingredients of Modified xiaoyao formula and their extracts,and to study transport rates of above elements. METHODS:Microwave digestion-in-ductively coupled plasma mass (ICP-MS) was used to determine above elements. Radio-frequency power was 1.50 kW,atomiza-tion pressure was 720 kPa,sampling depth was 7 mm,plasma gas flow rate was 15.0 L/min,carrier gas flow rate was 1.00 L/min, spray chamber temperature was 2℃,He flow rate was 4.5 mL/min under collision mode,measurement points/peak was 3,data ac-quisition mode was jump mode. RESULTS:The linear ranges of As,Pb,Cu,Hg,Cd,Mo,V,Cr,Mn,Fe,Ni,Zn,Ru,Rh, Pd,Os,Ir,Pt was 0-500.17 ppb(r was 0.9994-09999);LOD was 0.0004-5.3410 ng/g;RSD of repeatability tests was all lower than 5.0%;average recoveries were 82.0%-117.9%(RSD=0.2%-5.7%,n=9). Ru,Rh,Pd,Os,Ir,Pt,Hg had not been detect-ed. The transfer rates of 11 metals in the extracts ranged 1.46%-31.90%. CONCLUSIONS:The method is simple,precise,stable and repeatable,and can be used for simultaneous determination of 18 toxic elements in 10 ingredients of Modified xiaoyao formula and their extracts. The contents of toxic elements are decreased during the preparation of Modified xiaoyao formula extract to guar-antee the safety of drug use to certain extent.

Objective To study the distribution and correlation of 20 kinds of inorganic elements (Na,Mg,A1,V,Cr,Co,Fe,Ni,Cu,Zn,Mn,As,Se,Cd,Cs,Sr,Ag,Hg,Pb,and U) in Jiawei Xiaoyao Formula (JXF).Methods The levels of 20 inorganic elements in 10 kinds of herbal substances were determined by ICP-MS with microwave digestion for sample preparation,and the results were analyzed with SPSS22.0.Results There were some correlations among the inorganic elements in JXF;Four main factors were selected by principal component analysis (PCA).The PCA results showed that Na,Mg,A1,V,Ni,Cu,Se,Sr,Cs,Mn,Hg,and Fe might be the characteristic elements in JXF;There were Mg,A1,Mn,and Fe elements in 10 herbal drugs,the contents of inorganic elements in herbal drug were different because the medicinal parts were different,and the decreasing order was roots and rhizomes,skin type,all grass,fruit seeds,and fungi.Conclusion Contents of inorganic elements can not only characterize the stability of extraction process,and can provide experimental evidence for the study of traditional Chinese medicine compound compatibility law.

Objective To explore the effect of pre-treatment of subcutaneous injection of ketamine on remifentanil induced hyperalgesia and K+/Cl-cotransporter 2,KCC2) expression on spinal cord of rats.Methods60 male adult SD rats were randomly divided into five groups(n=12 in each group):control group (group C),the incision group(group I),the incision plus remifentanil group(group I+R),the incision plus ketamine group(group I+K) and the incision plus remifentanil and ketamine group(group I+R+K).Mechanical withdrawal threshold (MWT) was evaluated at 24 hours before incision(T0),2 hours,6 hours,24 hours and 48 hours after incision(T1~T4).The lumbar spinal cords of rats were taken out at T4 time point and the KCC2 detected was detected by immunofluorescence analysis and western blot analysis.ResultsCompared with group C(T1(14.5±1.7)g,T2(14.2±1.1)g,T3(13.9±1.8)g,T4(14.2±1.1)g),MWT of other groups at T1 (I(5.6±0.8)g,I+R(3.2±1.0)g,I+K(6.8±1.7)g,I+R+K(5.1±1.6)g),T2 (I(6.9±1.0)g,I+R(4.3±1.2)g,I+K(8.0±1.4)g,I+R+K(6.2±1.5)g),T3 (I(7.6±0.9)g,I+R(5.4±1.1)g,I+K(10.3±1.2)g,I+R+K(7.1±1.1)g),T4 (I(8.9±1.1)g,I+R(7.5±1.4)g,I+K(11.3±1.2)g,I+R+K(8.3±1.2)g)and the expression of KCC2 at T4 decreased (P<0.05).Compared with group I(T1(5.6±0.8)g,T2(6.9±1.0)g,T3(7.6±0.9)g,T4(8.9±1.1)g),MWT of group I+R (T1(3.2±1.0)g,T2(4.3±1.2)g,T3(5.4±1.1)g,T4(7.5±1.4)g) decreased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 decreased significantly (P<0.05).Compared with group I(T1(5.6±0.8)g,T2(6.9±1.0)g,T3(7.6±0.9)g,T4(8.9±1.1)g),MWT of group I+K (T1(6.8±1.7)g,T2(8.0±1.4)g,T3(10.3±1.2)g,T4(11.3±1.2)g) increased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 increased (P<0.05).Compared with group I+R(T1(3.2±1.0)g,T2(4.3±1.2)g,T3(5.4±1.1)g,T4(7.5±1.4)g),MWT of group I+R+K (T1(5.1±1.6)g,T2(6.2±1.5)g,T3(7.1±1.1)g,T4(8.3±1.2)g) increased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 increased (P<0.05).ConclusionPre-treatment of subcutaneous injection of ketamine can reduce the hyperalgesia of rats induced by remifentanil and reduce the inhibition of KCC2 expression on dorsal horn of spinal cord.

Objective To evaluate the role of spinal neuronal Mas-related gene receptor C (MrgC) in the maintenance of bone cancer pain (BCP) in mice.Methods A total of 132 SPF male C3H/HeJ mice, aged 8-10 weeks, weighing 18-22 g, were randomly divided into 4 groups (n=33 each) using a random number table: sham operation group (S group) , BCP group, bovine adrenal medulla peptide 8-22 (BAM8-22, a highly selective MrgC agonist) group (group BAM), and MrgC antibody group (group MA).BCP was produced by injecting α-MEM 20 μl containing 2×105NCTC2472 cells into the distal medullary cavity of right femur bone.While α-MEM 20 μl was injected only in group S.The artificial cerebrospinal fluid 5 μl was injected intrathecally in S and BCP groups, and BAM 8-22 8 nmol/5 μl and MrgC antibody 5 μl were injected intrathecally in BAM and MA groups, respectively, once a day for 7 consecutive days starting from the day 14 after inoculation of the tumor cells.At 1 day before inoculation (T0), before administration (T1) , and at 14, 16 19 and 21 days after inoculation (T2-5, at 0.5 h before the initial administration and 2 h after each administration) , the number of spontaneous flinches (NSF) and mechanical paw withdrawal threshold (MWT) were measured.Five animals selected from each group at each time point were sacrificed, and the lumbar enlargement segments of the spinal cord were removed for determination of MrgC expression in the spinal neurons (by immunofluorescence).Results Compared with group S, NSF was significantly increased, MWT was decreased, and the expression of MrgC was up-regulated at T1-5 in BCP, BAM and MA groups.Compared with group BCP, NSF was significantly decreased, MWT was increased, and the expression of MrgC was up-regulated at T2-5 in group BAM, and NSF was significantly increased, MWT was decreased, and the expression of MrgC was down-regulated at T2-5 in group MA.Conclusion Spinal neuronal MrgC is involved in the maintenance of BCP in mice.