Objective To investigate the role of glucocorticoid-induced tumor necrosis factor ligand (GITRL) in regulating the inflammatory reaction of kupffer cells.Methods The kupffer cells and T cells of mice were isolated and divided into 6 groups after being co-cultured:control group,kupffer cells and T cells were cultured in DMEM only; lipopolysaccharide (LPS) group,kupffer cells and T cells were cultured in media with LPS (1 mg/L) ; LPS + GITRL siRNA group,kupffer cells and T cells were cultured in media as the LPS group after transfected with GITRL siRNA ; LPS + control siRNA group,kupffer cells and T cells were cultured in media as the LPS group after transfected with control siRNA; LPS + pEGFP-N1 GITRL group,kupffer cells and T cells were cultured in media as the LPS group after transfected with pEGFP-N1 GITRL plasmid; LPS + pEGFP-N1 control group,kupffer cells and T cells were cultured in media as the LPS group after transfected with control plasmid.After 24 hours of treatment,the expressions of GITRL and PDL1 of kupffer cells were detected by immunofluorescence and western blot,respectively.The proliferation and apoptosis of T cells were measured by MTF assay and Annexin V/PI flow cytometry,respectively.The expression of tumor necrosis factor (TNF-α) in the supernatant fluid was measured by ELISA.All data were analyzed using the independent t test and one-way analysis of variance.Results The transfection efficiencies of GITRL siRNA and pEGFP-N1 GITRL were 90％ and 85％,respectively.Compared with normal kupffer cells,the protein expression of GITRL of kupffer cells transfected with GITRL siRNA was significantly decreased,while the protein expression of GITRL of kupffer cells transfected with pEGFP-N1 GITRL was significantly increased (t =41.72,13.10,P ＜ 0.05).There was no significant difference in the protein expressions of GITRL between normal kupffer cells and those in the control groups (F =2.27,P ＞ 0.05).The fluorescence intensity of GITRL in the LPS group was significantly higher than that in the control group (t =49.29,P ＜ 0.05).Compared with LPS group,the activation of GITRL expression by the LPS was significantly suppressed in the LPS + GITRL siRNA group (t =9.84,P ＜ 0.05),while the expression of GITRL in the LPS + pEGFP-N1 GITRL group was significantly increased (t =5.78,P ＜ 0.05).There was no significant difference in the GITRL expression among the LPS + control siRNA group,LPS + pEGFP-N1 control group and LPS group (F =0.86,P ＞ 0.05).The expression of PDL1 in the LPS group was significantly lower than that in the control group (t =18.83,P ＜0.05).Compared with LPS group,the expression of PDL1 in the LPS + pEGFP-N1 GITRL group was significantly suppressed (t =11.79,P ＜ 0.05),while the expression of PDL1 in the LPS + GITRL siRNA group was significantly stronger (t =19.08,P ＜ 0.05).There was no significantly difference in the expression of PDL1 in the LPS + control siRNA group,LPS + pEGFP-N1 control group and LPS group (F =2.22,P ＞ 0.05).The proliferation of T cells was increased and the number of apoptotic T cell was decreased in the LPS group when compared with control group (t =49.43,40.11,P ＜ 0.05).Compared with LPS group,the proliferation and apoptosis of T cells in the LPS + pEGFP-N1 GITRL group had the same trend (t =5.77,12.64,P ＜0.05); while the proliferation of T cells was decreased and the apoptosis of T cells was increased in the LPS + GITRL siRNA group (t =17.00,49.90,P ＜ 0.05).There was no significant difference in the proliferation and apoptosis of T cells among the LPS + control siRNA group,LPS + pEGFP-N1 control group and LPS group (F =1.87,1.35,P ＞ 0.05).The expression of TNF-α was significantly higher in the LPS group than that in the control group (t =125.68,P ＜ 0.05).Compared with the LPS group,the expression of TNF-α was significantly decreased in the LPS + GITRL siRNA group (t =119.65,P ＜ 0.05),while the expression of TNF-α in the LPS + pEGFP-N1 GITRL group was significantly increased (t =147.70,P ＜ 0.05).There was no significant difference in the TNF-α expression among the LPS + control siRNA group,LPS + pEGFP-N1 control group and LPS group (F =0.14,P ＞ 0.05).Conclusion Kupffer cells suppress the expression of PDL1 by upregulating GITRL,and thus activate the proliferation of T cells and promote the inflammatory reaction.The immunologic balance may be recovered after the interference of GITRL to restrain the inflammatory reaction.

Objective To observe the effects of storage condition on the quality of Semen Armeniacae Amarum. Methods Orthogonal design was adopted. The storage condition was optimized with temperature,humidity and storage time as the observation factors,with the content of amygdalin,and the content,acid value and peroxide value of fatty oil as the indexes.The content of fatty oil,acid value and peroxide value were measured according to the methods provided by 2005 edition of Chinese Pharmacopoeia. The content of amygdalin was measured by HPLC. Results Temperature was the main influencing factor of amygdalin content,then came humidity and storage time. Temperature was also the main influencing factor of the concent and the acid value of fatty oil,and the humidity was the second. For peroxide value,humidity was the main factor,then came temperature and storage time. Conclusion The optimal conditions of storing Semen Armeniacae Amarum as follows:storage in dry place with the temperature of 2～ 8 ℃,and in short storage time.

Objective:To explore the counselor verbal response modes of psychic hot-line counseling and its characteristics.Methods:Through a qualitative study with category analysis method,8 pieces of audio recordings were collected,in each of which the entire duration was between 30 minutes to 50 minutes.And all of these record-ings were transcribed word by word.Meaningful 1202 pieces were deeply analyzed.Results:There were 5 core counselor verbal response types in hot-line counseling,greeting response,set-up response,listening response,follow-up response,and affecting response,all of which had 24 concept categories.All of the online counselors used the 5 types of verbal response.The listening response and follow-up response were the highest frequent types.Greeting response and set-up response were used mostly in the initial stage of counseling.Listening response,and follow-up response were used mostly in the stage of describing problem.Affecting response was sharply increased in the stage of analyzing problem.Greeting response and set-up response were used mostly in last stage of counseling.Conclu-sion:It suggests that there are 5 core counselor verbal response types and 24 concept categories in hot-line counse-ling.The listening response and follow-up response are the highest frequent types.Different types of verbal re-sponse modes used by counselor in different stages due to different intention.

Objective:To observe the efficacy of acupuncture plus tuina therapy for insomnia.
Methods:Totally 100 eligible patients were randomized into a treatment group (n=51) and a control group (n=49) by using the random number table. The treatment group was intervened by acupuncture at Baihui (GV 20), Fengchi (GB 20), Wangu (GB 12), Shenting (GV 24), Anmian [Extra, locates at the midpoint between Yiming (EX-HN 14) and Fengchi (GB 20)], and Neiguan (PC 6) plus tuina treatment; the control group was intervened by acupuncture alone. For both groups, the treatment was given once a day, 10 sessions as a treatment course, and the efficacies were evaluated and compared after 3 treatment courses.
Results:Of the 51 subjects in the treatment group, 46 cases completed 3 treatment courses, and 17 cases were cured, 18 cases showed marked efficacy, 7 cases were improved, 4 cases had invalid result, and the total effective rate was 91.3%. Of the 49 subjects in the control group, 46 cases completed 3 treatment courses, and 8 cases were cured, 12 cases showed marked efficacy, 14 cases were improved, 12 cases had invalid result, and the total effective rate was 73.9%. The total effective rate of the treatment group was significantly higher than that of the control group (P<0.05).
Conclusion:Acupuncture plus tuina can calm the mind and activate meridian qi in treating insomnia, and it can produce a more significant efficacy than pure acupuncture treatment.

Objective To investigate whether polymorphisms of apolipopretein D gene (APOD) have an effect on the risk for sporadic Alzheimer's disease (SAD).Methods Combination of polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing technique to screen all exons (1-5),along with flanking exon-intron boundaries of the ApoD gene.We investigated the polymorphisms of ApoD in 256 SAD patients and 294 healthy controls from North China by PCR-RFLP technique.Association of every polymorphism with AD was analyzed in this case-control study.Results Two ApoD (rs5952 and rs1568566) polymorphisms were detected and there were significant differences in the genotype or allele frequencies for the 2 ApoD polymorphisms respectively between cases and controls.Logistic analysis showed that rs5952C or rs1568566T allele carrier increased the risk for SAD (rs5952 adjusted OR=1.817,95% CI 1.237--2.669,χ2=9.282,P=0.002 ; rs1568566 adjusted OR=1.563,95% CI 1.060-2.306,χ2=5.072,P=0.024).The APOD polymorphisms showed gender specific associations.The linkage disequilibrium of the 2 single nucleotide polymorphism loci was found between rs5952 and rs1568566 of ApoD.Conclusion Polymorphisms of rs5952 and rs1568566 in APOD might play an important role in modifying risk for SAD.

The development of formulas in the Ming Dynasty surpassed the previous dynasties in range and depth.The main achievements of formulas were not only the coming out of Puji Fang and Yifang Kao,but also the diversity of formulas nomenclature and variation,initial perfection of formulas classification and composition,continuous invention of new distinctive formulas,steady increment of remedy books and wide application of preparations.

Objective To study the upper airway structure characteristics of children with obstructive sleep apnea hypopnea syndrome (OSAHS) using electronic pharyngorhinoscopy.Methods Seventy-eight children with OSAHS (34 cases mild group,44 cases moderate and severe group) and 52 children with chronic pharyngitis in the same period (control group) were selected.Each group was examined by electronic pharyngorhinoscopy.The adenoid-nasopharynx ratio (ANR),degree of palatopharyngeal anteroposterior diameter collapse,degree of palatopharyngeal right-and-left diameter collapse,degree of palatopharyngeal cavity collapse,degree of glossopharyngeal anteroposterior diameter collapse,degree of glossopharyngeal right-and-left diameter collapse,degree of glossopharyngeal cavity collapse were compared,and the relevance between thegn were analyzed.Results The ANR in mild group,moderately and severe group was significantly higher than that in control group (0.84 ± 0.21 and 0.69 ± 0.25 vs.0.34 ± 0.12),and the ANR in moderate and severe group was significantly higher than that in mild group.There were statistical differences (P ＜ 0.05).The degree of palatopharyngeal anteroposterior diameter collapse,degree of palatopharyngeal right-and-left diameter collapse,degree of palatopharyngeal cavity collapse,degree of glossopharyngeal anteroposterior diameter collapse,degree of glossopharyngeal right-and-left diameter collapse,degree of glossopharyngeal cavity collapse in moderately and severe group were significantly higher than those in control group and mild group (0.81 ± 0.25 vs.0.41 ± 0.17 and 0.54 ± 0.20,0.66 ± 0.23 vs.0.31 ±0.08 and 0.43 ±0.14,0.51 ±0.18 vs.0.14 ±0.04 and 0.23 ±0.11,0.55 ±0.21 vs.0.29 ±0.14 and 0.32 ± 0.13,0.69 ± 0.28 vs.0.42 ± 0.12 and 0.43 ± 0.11,0.33 ± 0.12 vs.0.10 ± 0.03 and 0.14 ± 0.05),and there were statistical differences (P ＜ 0.05).There were no statistical difference in the indexes between mild group and control group (P ＞ 0.05).In children with OSAHS,the ANR,degree of palatopharyngeal anteroposterior diameter collapse,degree of palatopharyngeal cavity collapse,degree of glossopharyngeal right-and-left diameter collapse,degree of palatopharyngeal cavity collapse were positively correlated with AHI (P ＜ 0.05),while there was no correlation between the degree of palatopharyngeal right-and-left diameter collapse,degree of glossopharyngeal anteroposterior diameter collapse and AHI (P ＞ 0.05).Conclusions Upper airway structural changes in children with OSAHS have certain correlation with the severity of sleep apnea.And there is clinical value by using electronic pharyngorhinoscopy in children with OSAHS to research airway morphology.

ObjectiveTo investigate the relationship between the polymorphisms of the promoter of a disintegrin and metalloproteinase 10(ADAM10) gene and sporadic Alzheimer's disease (SAD).Methods The promoter of ADAM10 gene in 10 controls and 10 SAD patients was sequenced.Three variations were found,then these variations in 298 SAD patients (SAD group) and 315 healthy controls (control group)were genotyped by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).ResultsThree polymorphisms were found in the promoter of ADAM10 gene: -279G/A (rs653765),- 630G/T( rs514049 ) and - 921GAGA/- ( rs33926666 ).For - 921GAGA/-,there were significant differences in genotype ( GAGA/GAGA:138 (46.3％ ),GAGA/-:155(52.0％),-/-:5(1.7％))and allele frequencies (GAGA:431 (73.6％),-:165 (27.7％) ) between SAD and control (genotype:x2 =34.130,P =0.000; allele:x2 =25.972,P =0.000). For - 279G/A,there were significant differences in genotype and allele frequencies between SAD and control in the subjects without ApoEε4 allele (genotype:x2 =8.734,P=0.013; allele:x2 =5.129,P=0.024). -279G and -921GAGA were relatively protective allele types for SAD,and they were not in linkage disequilibrium.ConclusionThe polymorphisms - 279G/A and - 921GAGA/- of ADAM10 are associated with SAD.Allele G or genotype G/G of -279G/A and the GAGA/GAGA genotype or the GAGA allele of -921GAGA/- might have a protective effect on SAD.

Objective To compare the effect of two different time for intravenous dexmedeto-midine alleviates shivering during parturients cesarean undercombined spinal-epidural anaesthesia. Methods One hundred and twenty parturients delivery under combined spinal-epidural anaesthesiawere equally randomized into three groups.The parturients were intravenously injected with dexmedetomidine 0.4 μg/kg (15 min)at 15 min before the beginning of anesthesia (group A)or the time immediately after delivery of fetus (group B)respectively.Moreover,group C was the control group.Parturientsin group C were given equal volume of normal saline.Mean arterial pressure (MAP),heart rate(HR),pulse oxygen saturation (SpO 2 )and ear temperature were recorded respec-tively.Apgar scores of the fetus at 1 min and 5 min,the incidence and grade of shivering response, Ramsay sedative grade and adverse responses such as nausea,vomiting and traction reaction were re-corded as well.Results By 30 min after operation,2 parturients (5%)in group A,8 parturients (20%)in group B and 19 parturients (47.5%)in group C experienced shivering.Traction reaction was observed in 2 parturients (5%)in group A,in 3 parturients (7.5%)in group B and in 10 parturi-ents (25%)in group C respectively.Compared with group C,the incidence rate and degree of shive-ring were significantly lower in groups of A and B (P <0.05),as well as the traction reaction (P <0.05).The incidence rate and degree of shivering in group A were significantly lower than that in group B (P <0.05),but the incidence rates of traction reaction were not statistically different between the two groups.Conclusion Dexmedetomidine administered before combined spina-epidural anesthesia could prevented shivering response in parturients undergoing cesarean section better than being injected immediately after the birth offetus.

Objective To analyze the clinical and pathological features of lipid storage myopathy (LSM) caused by primary carnitine deficiency (CD).Methods The clinical data of 4 cases of possible LSM caused by primary CD were analyzed retorspectively.Results The clinical features of the 4 patients were subacute or chronic onset, proximal muscle weakness and exercise intolerance. Elevated levels of creatases were measured in serum and myogenic damage was found by EMG examination. Frozen sections of muscle biopsy samples showed many fibers contained numerous vacuoles which was stained by oil red O. No ragged red fiber (RRF) was seen in MGT stain. Type I fibers were more severely affected. In electron microscopy, the prominent abnormality was the presence of excessive amounts of fatty droplets in muscle fibers and subsarcolemmal regions with mild increased mitochondria.Treatment with glucocorticoid and energy supplement had been clinically beneficial. Conclusions Fatigue and muscle weakness are prominent manifestations in LSM caused by primary CD. The main changes are accumulation of lipid droplets in muscle specimen without prominent abnormality in structure in mitochondria. Good clinic effect may be caused by therapy with glucocorticoid.