Objective To study the localization of specific allergen of Dermatophagoides pteronyssinus. \ Methods\ Through optical microscope,the specific allergens of D.pteronyssinus were observed in paraffin sections using D.pteronyssinus\|specific IgE antibodies from the patient sera. \ Results and Conclusion \ The digestive system was found occupying large parts of body cavity of D.pteronyssinus by HE staining, while the specific allergens of D.pteronyssinus were mostly occurred in the midgut tissue, gut contents, cuticle and reproductive system in the immunostained sections. The results also showed that many parts of D. pteronyssinus were recognized by the specific IgE antibodies obtained from allergic individuals to D.pteronyssinus, which provided a theoretic base for further study of isolation and purification of the specific allergen.

Objective To study the value of serum procalcitonin(PCT) to the early diagnosis of sepsis .Methods From June 2014 to June 2015 ,a total of 686 cases were enrolled in this retrospective study .PCT tests were assayed within 2 days of bacterial culture .Results In this study ,56 cases ,67cases ,and 567cases were classified into the positive blood culture group ,positive body fluid culture group ,and negative all culture group ,respectively .Median PCT values were 4 .26 2 .78 ,0 .46 ng/mL ,respectively .Me-dian PCT values in the gram-positive bacterial culture group and gram-negative bacterial culture group ,respectively ,were 2 .35 and 4 .56 ng/mL .Median PCT values in the positive hydrothorax culture group ,positive ascites culture group ,and positive bile culture group ,respectively ,were 1 .91 ,5 .23 ,3 .64 ng/mL .In all ,Median PCT values of 47 cases of sepsis and 16 cases of severe sepsis were 5 .32 and 10 .25 ng/mL ,respectively .Conclusion PCT level is correlated with the severity of sepsis ,pathogenic bacteria type ,and the site of infection ,and can be used in the early diagnosis of sepsis .

This review introduced the research progress of covalent modification strategies in local anesthetic drug delivery systems. As a commonly used and multimodal analgesic drug, local anesthetics have limited duration of action and potential toxicity in clinical application. In order to prolong the analgesic effect and reduce systemic toxicity, researchers are committed to the development of sustained-release local anesthetics with long-lasting dose-controlled-release functions. When it comes to the delivery of local anesthetics, the covalent modification strategy is a key approach. By covalently binding drugs to large molecule carriers, covalent modification strategies can improve drug stability, targeting and delivery efficiency. Macromolecular prodrugs can modulate the kinetic process of the drug, so that the drug is released in the form of the active ingredient and achieve better therapeutic effects. In recent years, stimulus-responsive macromolecular prodrugs have become a research hotpot for local anesthetic drug delivery systems, and the stimulus-responsive performance of macromolecular prodrugs can rapidly release drugs under internal and external stimulus conditions, and maintain low toxicity and high efficiency in blood circulation and normal tissues. These emerging research directions provide important guidance for prolonging the analgesic effect of local anesthetics and reducing systemic toxicity, and provide new idea for the development of more effective drug delivery systems in the future.

Objective To observe the in vitro anti-Coxsackievirus B3m (CVB3m) effect of sophocarpine(SC) extracted from Sophora flavescens, a traditional Chinese herb. Methods HeLa cells were cultured and the micro-dose cytopathic effect (CPE) assays were applied to detect the toxicity of SC. CPE-inhibitory assays were used to observe the in vitro anti-CVB3m effect of SC. MTT and crystal assays were introduced to examine the anti-CVB3m effect of SC. HeLa cells were infected with CVB3m and added with SC in different concentrations 15 h later.The viability and number of survival of HeLa cells were determined by MTT and crystal violet assays, respectively. Results No toxicity was found on HeLa cells by SC with concentrations 100 ?g/mL, SC could accelerate and aggravate the CPE. SC could protect the CVB3m-infected HeLa cells with concentrations from 1.56 to 25 ?g/mL, and the viability and cell number measured by MTT and crystal violet assay in the SC-handled cells were higher and bigger than those in the virus infected ones. However, the inhibitory effect of virus was exacerbated with higher concentrations (50 and 100 ?g/mL), and the cell number and viability of the SC-handled cells were smaller and lower than those of the infected ones. Conclusion SC with a proper concentration has the in vitro anti-CVB3m effect and may protect HeLa cells from CVB3m infection.

Objective To investigate the possibility and utility of metformin alone or in combination with fosinopril to reduce blood pressure in patients with essential hypertension.Met hods A total of 140 cases of non-diabetic essential hypertension with hyperinsulinemia were recruited and randomly assigned to two groups: a group of 68 treated with metformin 500 mg tid and a group of 72 treated with fosinopril 10 mg qd.The duration of the treatment was 8 weeks.Combination therapy with the two drugs was used after 4 weeks of treatment if needed.If the target goals of systolic blood pressure (SBP) < 140 mm Hg (1 mm Hg =0.133 kPa) and /or diastolic blood pressure (DBP) <90 mm Hg were not attained 4 weeks, combination therapy with two drugs was used in either group in the next 4 weeks.The changes of blood pressure and insulin sensitivity of the two groups were observed before and after treatment.Results (1) After 4 weeks of treatment, SBP in metformin group and fosinopril group decreased by ( 13.0 ± 1.2) mm Hg and (15.4 ± 1.4) mm Hg, and DBP decreased by (9.0 ± 1.0) mm Hg and ( 10.4 ± 1.1 ) mm Hg respectively.After 8 weeks of treatment, SBP in metformin group and fosinopril group decreased by (17.8 ± 1.5) mm Hg and (20.9 ± 1.5) mm Hg, and DBP decreased by (13.2 ±0.9) mm Hg and (15.3 ± 1.1) mm Hg respectively.There was no significant difference in the decline of blood pressure between the two groups (P >0.05).The rates of combination therapy were both 54% in the two groups.(2) Fasting insulin as well as 30 min and 120 min insulin levels after oral glucose tolerance test and insulin area under the curve in the metformin group were significantly reduced after 4 and 8 weeks of treatment as compared with those of baseline(P < 0.05 and P < 0.01 ) .In the fosinopril group, however, they decreased only after 8 weeks treatment (P < 0.05).The insulin action index in the metformin group was higher than that in the fosinopril group after 4 weeks of treatment (P <0.05) ,but there was no significant difference between the two groups after 8 weeks of treatment (P > 0.05).Conclusion Metformin and fosinopril have similar antihypertensive effect and a good synergy in essential hypertension with hyperinsulinemia.

Aim To investigate the effects of 6% hydroxyethyl starch 130/0.4(6%HES 130/0.4)and Ringer's solution resuscitation on early lung injury in hemorrhagic shock rats and its mechanisms.Methods Twenty-four male SD rats were divided into 4 groups:sham,Ringer's solution(RS),two HES groups(H1,H2).Group H1,H2 received HES 33,50 ml·kg~(-1) and Ringer's solutions respectively after 90-minute shock(the dose of Ringer's solutions was 3 times as muchas the maximum shed blood volume minus the dose of HES).Blood samples were taken from artery for blood analysis and the expression of CD11b/CD18 at T_0,T_1,T_4,T_5.The lungs were removed for ultrastructure examination.Results PaO_2 increased in group H1 at T_(1～5) and group H2 at T_5 as compared with T_0.PaCO_2 decreased in all the resusitation groups.The ultrastructure was basically normal except that the mitochondria changed slightly in group H1.In group H2,the perinuclear space was dilated slightly and the rough endoplasmic reticulum expanded slightly,and the degranulation was observed.Compared with group RS,the expressions of CD11b and CD18 decreased at T_4,T_5;compared with group H1,the expressions of CD11b and CD18 in group H2 increased.Conclusions Treatment with 6%HES(130/0.4)33 ml·kg~(-1)and Ringer's solution can attenuate hemorrhagic shock,and the resuscitation reduces lung injury through inhibition of expressions of CD11b and CD18.

Objective:To express recombinant protein mIL-21-hIgGFc in 293E cells,and investigate its effect on CD8+T cell.Methods:Total RNA was extracted from the mouse spleen cells ,and then IL-21 gene was amplified by RT-PCR and inserted into expression vector PTT3-hIgGFc.PTT3-mIL-21-hIgGFc were transfected into 293E cells by calcium phosphate method.The supernatants were collected at 48 hours and 72 hours and concentrated by MOLLIPORE Labscale TM TFF system ( 5 kD membrane ).The mIL-21-hIgGFc fusion protein was purified with HiTrap TM Protein G column.The protein was quantified by SDS-PAGE and ELISA.The biological activity of the protein was determined by detecting the change of the phenotypes of CD 8+ T cells treated with the protein.Results: The constructed recombinant plasmid PTT 3-mIL-21-Fc was confirmed by sequencing.PTT3-mIL-21-Fc was transfected into 293E cells,mIL-21-Fc protein in culture supernatant was collected after 48 hours and 72 hours.The protein in cell su-pernatant reached a concentration of 787 ng/ml which was determined by ELISA.The protein was purified by Protein G chromatography column.P1A-specific T cells were treated with mIL-21-hIgGFc, and found that the CD44low CD62Lhi CD8+ population increased compared to the control.Conclusion:We built PTT3-mIL-21-hFc recombinant plasmid, expressed mIL21-hFc fusion protein in 293E cells,and purified by Protein G column.By treating mIL-21-hFc ,the antigen-primed CD8+T cells prefer to differentiate into CD44low CD62Lhi CD8+T cells which had been reported as a memory stem phenotype .This protein may be used to improve the effectness of adoptive T cell cancer therapy.

OBJECTIVE To investigate the bacterial infection and drug susceptibility of 91 victims wounded in Wenchuan earthquake from Sichuan and provide the laboratory evidences for sequential treatment.METHODS The specimens such as sputum,wound secretion,urine,blood,stool and necrotic tissue from the wounded were cultured on different kinds of culture medium under the aerobic or the anaerobic condition.The isolated bacteria were identified by VITEK GN analyzer or GP cards and API 20C analyzer for fungus.The drug susceptibility tests were carried out by the K-B method.RESULTS 91 wounded(58.7%) were infected,most of them were more than 60 years old.Respiratory tract(49.5%),wound(30.0%) and urinary tract(6.6%) were the main infectious sites.Among 91 strains of isolated bacteria,56 strains were G-bacilli(61.5%),20 were G+ bacteria(22.0%) and 12 strains were fungi(13.2%).The most common species among them were Escherichia coli(13.2%),fungi(13.2%),Acinetobacter(11.0%),Burkholderia cepacia(11.0%),Enterococcus(9.9%) and Staphylococcus aureus(9.9%).Drug susceptibility results showed that except the Pseudomonas aeruginosa and B.cepacia,the most G-bacilli were sensitive to imipenem,but resistant to the first and second-generation cephalosporins.The most G+ cocci were sensitive to VAN,LNZ and QDA antibiotics,and the fungus were sensitive to antibiotics used commonly in clinical treatment.CONCLUSIONS The pathogenic culture and the drug susceptibility analysis of the victims wounded in Wenchuan earthquake are inportant to guide the anti-infectious treatment.

Objective To identify serum biomarkers for rheumatoid arthritis(RA)by protein finger- print pattern. Methods One hundred and forty-one serum samples of 90 RA patients, 20 systemic lupus ery- thematosus(SLE)patients, and 31 healthy individuals were randomly divided into training set(n=93, 60 RA patients, 13 SLE patients and 20 healthy individuals)and test set(n=48, 30 RA patients, 7 SLE patients and 11 healthy individuals). They were detected by surface enhanced laser desorption/ionization-time of flight- mass spectrometry(SELDI-TOF-MS). The protein fingerprint pattern obtained from SELDI-TOF was trained by a multi-layer artificial neural network(ANN)to establish a diagnostic model. Results The detective mod- el obtained by ANN was used to detect the 48 unknown serum samples. The sensitivity and specificity for RA detection was 90% and 90.9% respectively. Conclusion In comparison with traditional methods, SELDI- TOF-MS could identify new serum biomarkers in RA. Combined with ANN, it provides high sensitivity and specificity for RA diagnosis.