Objective:To establish a method for the content determination of total nitrogen in Nidus Vespae by salicylic acid and sodium hypochlorite spectrophotometry. Methods:The content of total nitrogen in Nidus Vespae from different areas with various batches was detected. Nidus Vespae was treated with sulfuric acid and hydrogen peroxide, and colored by salicylic acid and sodium hypochlo-rite. Using ammonium chloride as the control sample, the content of total nitrogen in the samples was tested at 650 nm. To ensure the method reliability, single factor experiments were performed to study the effects of temperature, reaction time and reagent dosage; or-thogonal experiments were used to screen optimum amounts of phosphate buffer solution, sodium salicylate solution and sodium hypo-chlorite solution. Results:The results showed that the optimum temperature was ambient, and the optimum time was 1. 5h. The opti-mum amount of phosphate buffer solution , sodium salicylate solution and sodium hypochlorite solution was 4 ml,7 ml and 1. 5 ml, re-spectively. The linear range of total nitrogen was 0. 06-0. 78 μg·ml-1(r=0. 9998), and the average recovery was 100. 98% (RSD=2. 10%,n=6). Conclusion:The method can be used to determine the total nitrogen content in Nidus Vespae. The total nitrogen content in Nidus Vespae is various in various areas with different batches.

Objective:To explore the correlation among the expression of p53, p21, and PCNA in the negative surgical margin of early laryngeal cancer and its relationship with local recurrence of the cancer. Methods:Data of 92 patients with early laryngeal cancer admitted to the Tangshan Union Hospital from 2004 January to 2010 December were selected. Immunohistochemical method was used to detect the expression of p53, p21, and PCNA in the negative surgical margin tissues of cancer. All patients were followed up for two years to observe survival status and tumor recurrence. Results:The expression of P53, p21, and PCNA had no differences in the cancer tissues in terms of grade and stage. Two years after the surgery, 16 of the 92 patients with laryngeal cancer presented recurrence with a rate of 17.39%. The recurrence rates of expression of p53, p21 and PCNA protein in the positive cut edge were 50.00%, 34.21%and 33.33%, significantly higher than those of negative ones (8.33%, 5.56%and 9.68%). The positive rates of the three proteins were higher in the recurrence specimen than in the non-recurrence while there was a significant correlation between expression of PCNA and p21 (P<0.01). The expression of both p21 and p53 in laryngeal carcinoma was also significantly correlated with the negative margin. Con-clusion:The expression of p53, p21, and PCNA in the surgical margin of early laryngeal cancer are valuable biological markers for the prognosis of these patients. Thus, combined detection of p53, p21, and PCNA in laryngeal margin tissues can be used as a predictive in-dicator of the recurrence.

Purpose To compare the therapeutic effects of CO2 laser resection and traditional split laryngeal surgery on the early laryn-geal cancer, and to examine relationship between the expression of p27 and PTEN and clinical prognosis in early laryngeal squamous cell carcinoma. Methods 85 patients who underwent CO2 laser resection surgery and 46 patients who underwent split laryngeal surger-y were selected and the occurrence rates after surgery were observed. SP immunohistochemical method was uesd to detect the expression of p27 and PTEN in tumor resection marginal and their surgical margine tissues. Clinical data were collected and all patients were fol-lowed up. Results In the more than two-year follow-up study, 14 of 85 patients in CO2 laser treatment group ( recurrent group) pres-ented with local recurrence and the recurrence rate was 16. 5%, while 6 of 46 patients in split laryngeal surgery group presented with local recurrence and the recurrent rate was 13. 0%. There was no statistical significance in recurrence rate between the two groups ( P> 0. 05). 10 of all the none recurrent patients did not follow the doctor’s advice to quit smoking after the operation, while 12 in the recurrent patients did not, the difference between the two groups was statistically significant (P<0. 01). The positive rate of p27 and PTEN in laryngeal carcinoma tissues and the cancer adjacent tissues ( negative surgical margin tissues ) was 43. 5% ( 57/131 ) , 80. 2% (105/131) and 48. 9% (64/131), 83. 2% (109/131), respectively, with a significant difference (P<0. 01). The positive rate of p27 and PTEN in laryngeal carcinoma tissues of the recurrent group and non recurrent group was 20. 0% (4/20), 47. 7% (53/111) and 10. 0% (2/20), 55. 9% (62/111), respectively, with a significant difference (P<0. 05). While the positive rate of p27 and PTEN in tumor resection marginal tissues of the recurrence group and non recurrence group was 50. 0% ( 10/20 ) , 85. 6% ( 95/111) and 40. 0% (8/20), 91. 0% (101/111), the difference was also statistically significant (P<0. 01). Conclusions There was no statistically significant difference in tumor recurrence rate between CO2 laser surgery and traditional split laryngeal surgery. Postoper-ative recurrence is closely related to resume smoking. The recurrence rate of p27 and/or PTEN negative patients was higher than that of the opposite ones which should be followed up closely after treatment.

Purpose To study the expression of p53, p21 and Cdk1/p34cdc2 in the laryngeal cancer and its margin tissues and to ex-plore their relationship with local recurrence of laryngeal cancer. Methods A total of 85 patients with early laryngeal cancer were se-lected randomly during 2004 to 2010 in Tangshan Union Hospital, Hebei, China. SP immunohistochemical method was used to detect the expression of p53, p21 and Cdk1/p34cdc2 in the tumor and margin tissues. Pathological data were collected for follow-up. Results In more than 2 years of follow-up study, 14 of 85 patients with laryngeal cancer presented with recurrence (recurrent group), while 71 patients without recurrence (none recurrent group). The positive rate of p53 protein in laryngeal cancer and its margin tissues was 60. 0% and 36. 5%, respectively, the positive rate of p21 protein in laryngeal cancer and its margin tissues was 38. 8% and 21. 2%, respectively. The positive rate of Cdk1/ p34cdc2 in laryngeal cancer and its margin tissues was 70. 6% and 29. 4%, respectively. p53 protein in the surgical margin of the recurrent group and non recurrent group was 71. 4% and 29. 6% (P = 0. 003), that of p21 was 50. 0% and 15. 5%, (P =0. 004) and Cdk1/ p34cdc2 was 57. 1% and 23. 9% (P =0. 013), respectively. There was no correlation between expression of p53 with p21 protein and Cdk1/ p34cdc2 protein(P > 0. 05). Conclusion p53, p21 and Cdk1/ p34cdc2 may be involved in the occurrence, development and recurrence of laryngeal squamous cell carcinoma. Overexpression of p53, p21 and Cdk1/ p34Cdc2 in the surgical margin is closely related to local recurrence of laryngeal cancer.

Objective:To investigate the expressions of peroxisome proliferator-activated receptor gamma (PPARγ), estrogen re-ceptor alpha (ERα), and estrogen receptor beta (ERβ) in endometrial carcinoma and to analyze their correlations and clinical signifi-cance. Methods:Immunohistochemical assay and Western blot were used to detect the expressions of PPARγ, ERα, and ERβin normal endometrial tissues and well-differentiated, moderately differentiated, and poorly differentiated endometrial carcinomas. Results:PPARγexpression was significantly lower in endometrial carcinoma than in the normal endometrium and was intimately associated with cli-ni-copathologic variables. ERαexpression gradually decreased in moderately and poorly differenti-ated endometrial carcinomas. How-ever, no significant differences were found between the normal endometrium and well-differentiated endometrial carcinoma. ERβex-pression only decreased in the poorly differentiated endometrial carcinoma. No significant association was observed between ERβand clinicopathologic variables. Pearson correlation analysis showed a significant positive cor-relation between the expressions of PPARγand ERα. No correlations were observed between the expressions of ERαand ERβand between that of ERβand PPARγ. Conclusion:The expression lev-els of PPARγand ERαwere significantly associated with the clinicopathologic stage of endometrial carcinoma, and have essential functions in endometrial tumorigenesis and tumor progression.

Rehabilitation therapeutics has been put in practice in China. The setting of clinical course needs to be further explored. Thispaper summed up the past experience of diagnostic teaching by questionnaire for students to guide future teaching.

Although DNA vaccination is now a promising strategy against hepatitis B virus (HBV) infection, this approach has relatively modest antiviral effect, indicating that immunosuppressive mechanisms may occur in the long-term established infection. In this study, we studied the immunogenicity and anti-HBV efficiency of a combination of HBV surface (HBsAg) and core (HBcAg) DNA vaccine, enhanced by heat shock protein (HSP) gp96 or HSP70 and mediated by in vivo electroporation. Immunization with gp96 adjuvanted HBsAg/HBcAg DNA formulation induced potent T cell and antibody immunity against HBsAg and HBcAg. Notably, treatment with gp96 or HSP70 as adjuvant resulted in reduction of Treg populations by around 20%. Moreover, compared with nonimmunized control mice, immunization with gp96 or HSP70 adjuvanted DNA vaccine dramatically decreased serum HBsAg and viral DNA levels, and HBcAg expression in liver. These results may therefore provide an effective strategy for designing gp96-based DNA vaccine for immunotherapy of chronic HBV infection.
Adjuvants, Immunologic ; Animals ; Electroporation ; HSP70 Heat-Shock Proteins ; immunology ; Hepatitis B Core Antigens ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; immunology ; Hepatitis B, Chronic ; prevention & control ; Immunization ; Membrane Glycoproteins ; immunology ; Mice ; Mice, Transgenic ; Vaccines, DNA ; immunology

Objective To identify immunodominant B linear cell epitopes in capsid proteins VP1 to VP3 of Coxsackievirus A16 (CVA16) strain YY157.Methods The protean algorithms of bioinformatic software Lasergene were used to analyze antigenicity, hydrophilicity and surface probability of amino acid sequence of CVA16 capsid proteins VP1 to VP3.Multiple regions containing potential lineal B cell epitopes were predicted and their corresponding average indexes were calculated by BepiPred 1.0 Server.Corresponding peptides were synthesized and examined in peptide-enzyme-linked immunosorbent assay (ELISA) individually to check whether it reacted positively or negatively toward sera from children with confirmed CVA16 infection.Results Totally 21 possible B cell linear epitopes were predicted according to their relatively strong antigenicity, hydrophilicity and surface probability.The corresponding synthetic peptides reacted positively with sera of CVA16-infected children in a varying extent.Conclusion Immunodominant B cell linear epitopes of capsid proteins VP1 to VP3 of CVA16 strain YY157 are successfully predicted and confirmed.

Objective To evaluate expression and significance of TLR4 and NF-κB on inflammatory injure after intracerebral hemorrhage in rats .Methods 60 Sprague Dawley maleness rats were randomly divided into Sham group ,12 h ,24 h ,72 h and 7 d af-ter ICH group(12 s) .The ICH was induced by injection of autologous blood in rats .The behavioral changes were detected by neu-rologic deficit score .The water content of the brain was used to evaluate brain edema changes .Number of TLR4 and NF-κB positive cells by Nissl staining and the expression of protein determined by immunohistochemistry and Western blot .Results After ICH 12 h ,expression of TLR4 and NF-κB positive cells around the hematoma were expressed ,with the extension of the time ,expression was gradually increasing ,and after ICH 72 h the expression of protein were the highest .Cerebral edema and severe neurological damage occurred .Western blot shows the amount of TLR4 expression and NF-κB were in line with the result .Conclusion After in-tracerebral hemorrhage in rat causing inflammatory injure of brain tissue around the hematoma .TLR4 may activate the expression of NF-κB involved in the secondary inflammatory injure after intracerebral hemorrhage in rats .

OBJECTIVETo compare the clinical efficacy difference in dysantonomia between transcutaneous electrical stimulation at Renying(ST 9) combined with stellate ganglion block(SGB) and simple SGB.

METHODSSixty patients in accord with the diagnostic criteria of dysantonomia were randomly divided into two groups,30 cases in each group. In the observation group,transcutaneous electrical stimulation at Renying(ST 9) combined with SGB were adopted; in the control group,simple SGB was applied. In the two groups, treatment was used three times a week,and nine treatments were considered as one course. There was an interval of one week between courses,and two courses were treated. Total seven weeks were required. Scores were evaluated according to subjective symptoms before treatment,one month and three months after treatment in the two groups.

RESULTSThe scores of subjective symptoms were not statistically different before treatment in the two groups(P>0. 05). The scores of subjective symptoms one month and three months after treatment were all lower than those before treatment(all P< 0. 01), and subjective symptoms scores in the observation group were lower than those in the control group(both P<0. 01).

CONCLUSIONTranscutaneous electrical stimulation at Renying(ST 9) combined with SGB could obviously enhance the clinical effects for dysantonomia, and the control and improvement for clinical symptoms are apparently superior to simple SGB.

Acupuncture Points ; Adult ; Anesthetics ; administration & dosage ; Autonomic Nerve Block ; Autonomic Nervous System Diseases ; drug therapy ; therapy ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Stellate Ganglion ; drug effects ; physiopathology ; Transcutaneous Electric Nerve Stimulation ; Young Adult