Objective: To present the clinical experience in treating thymomas and thymomas with myasthenia gravis (MG). Methods: Between 1975 to 1997, 258 patients with thymoma were surgically treated. 168 cases of thymus tumor (TT, Group I) were compared to 90 cases of thymus tumor with myasthenia gravis (TTMG, Group II). Results: In group I, the average age was 38.4 years, the youngest was 4 years old. In 93.4%, the diameter of tumors was 5cm. 61.2% were in stages III and IV of clinical pathology. In group II, the average age was 46.2 with the youngest of 20 years old, the diameter of tumor under 3cm and 5cm was 34.6% and 65.6%. 55.2% was in stage I of clinical pathology. Conclusion: Many cases of thymoma in early stage received early treatment. The diagnostic standard of thymoma in early stage is: (1) the diameter of tumor is below 3cm. (2) In stage I of clinical pathology. The clinical characteristics of TTMG are : short history, the rapid progress, serious MG symptoms, and the high incidence of thymus-crisis.

To explore indications regularity of analogous decoctions of Erchen Decoction.323 analogous decoctions of Erchen Decoction were found in Concise Thesaurus of TCM Formula.Diseases and symptoms of TCM were analyzed.22 diseases and 118 symptoms of TCM were found.The main indication of analogous decoctions of Erchen Decoction included spleen-stomach and lung system,other symptoms were broadly involved.Erchen Decoction was the most appropriate for phlegm.

The double-window technique was applied in 30 patients with thoracic diseases in CT examination. The pictures of lung window, mediastinum window and double-window were read respectively and the diagnostic accuracy were compared. The principle, advantage and limitation of double-window technique were discussed.

Objective To summarize the experience of surgical management of carotid tumors ( CBT ) and application of shunt between common and internal carotid artery intraoperatively. Methods Thirty patients of CBT (mean age:39. 2 ±2. 3 years old,10 male and 20 female, 15 in left, 14 in right and 1 in both sides) who underwent surgical resection, were retrospectively reviewed. The average size of CBT was 4.9 ±0.3 cm. The diagnosis was established by ultrasound, CT, MRI or carotid arteriography. 16 patients underwent surgical resection of CBT, 10 patients underwent additional ligation of external carotid artery, and 4 patients underwent additional intraoperative shunt between common and internal carotid artery. Results Surgical procedures were successfully performeded in all 30 patients with CBT. Intraoperative shunts were successfully used between common and internal carotid artery in 4 patients. The postoperative complications included hoarseness (15) , bucking (11), crooked tongue ( 17) , dyspnea (1), dysphagia(3). There was no hemiplegia and death. Conclusion Surgical resection is the choice of treatment of carotid body tumor. The application of intraoperative shunt between common and internal carotid artery in complicated Shamblin Ⅲ stage is safe and effective.

0.05). ③The entrance point of screw was designed based on the obtained data. Sixty screws were all inserted into pedicles successfully. CONCLUSION: L1-5 superior margin root of spinous process locates in superior and inferior planes of vertebral pedicle with stable anatomic relationship; in addition, there are no significant differences between plain tomography and CT films. It provides anatomic data for pedicular operation.

AIM:To explore a method of isolation,purification,culture and identification of human microglia.METHODS:The brain tissue from abortive fetus was sterilely obtained,then chopped and triturated gently.The suspension was filtered and centrifuged to separate and isolate mixed glia cells.The cell density was adjusted to 2?1010 cells/L with DMEM/F12 complete medium and cultured in 75 cm2 flask at 37 ℃ in CO2 incubator(marked as first 0 d).After 7-10 days culture,floating cells including microglia and oligodendrocytes appeared in the flask.The first time purification was carried out to obtain highly purified microglia,for which the flasks were shaken gently.The floating cells were collected and transferred into a new flask(marked as second 0 d).4-5 days later,when microglia and oligodendrocytes grew in confluent state,the second purification was performed,for which the mixed cultured cells were digested with trypsin-EDTA and cell suspension was centrifuged,then the cell pellet was suspended by DMEM/F12 complete medium and cultured in flask(marked as 0 h).After 2 h,non-microglial cells including oligodendrocytes and cell debris were removed,and fresh medium was added into the adherent cells.In this way,the whole procedure of microglia purification was completed and the highly purified microglial cells were obtained.After purification,the expression rate of CD45,CD11b and CD68 on/in microglia were detected by laser-confocal microscopy and flow cytometry on the basis of immuno-fluorescence staining to identify the purity of microglial cells.Microglial phagocytotic function was evaluated by phagocytosis of fluorescent microspheres.The level of activation was identified by the phenomena of membrane ruffling in combination with fluorescent phalloidin staining.RESULTS:More than 98% of the microglias that we isolated and purified expressed CD45,CD11b and CD68.Almost all of the microglias could phagocytize fluorescent microbeads CONCLUSION:We succeed in isolating and purifying human microglias,which express CD45,CD11b and CD68.Almost all of the cells exhibit phagocytic function.So these cells will be useful for further functional and proteomic studies.

Objective To examine the effects of p38 mitogen-activated protein kinase (MAPK) inhibitor on the behavioral response to zebrafish larvae after hypoxia/reoxygenation brain injury and to identify whether the protective effect is mediated by inhibiting apoptosis and protein,and mRNA related to apoptosis.Methods The 5-day post-fertilization zebrafish larvae were randomly assigned to 3 groups:control group,model group and intervention group.Fishes in the intervention group were separated into 3 subgroups according to p38 MAPK inhibitor concentration (5,10,20 μmol/L).The activity levels of the larvae were analyzed by using quantization mode of ZebraLab software,swimming distance and moving speed were recorded.Terminal transferase dUTP nick end labeling (TUNEL) assays of brain assays were performed.The protein levels of phosphorylation of p38 MAPK,apoptosis related proteins of B-cell lymphoma-2 (Bcl-2),Bcl-2 associated X protein(Bax) and Caspase-3 were determined by Western blot.The mRNA expressions of Bcl-2,Bax,and caspase-3 were also analyzed by reverse transcription-quantitative PCR (RT-qPCR).Results The activity movement analysis of the intervention group 2 and 3 demonstrated a significantly increase in the swimming distance compared with the model group(P ＜ 0.05).After irradiation under strong light,all groups showed dramatically increasing in the moving speed.After removal of strong light,a significant decrease in moving speed was found in the control group and intervention group 2 and intervention group 3.The TUNEL assay showed that apoptosis index decreased in the intervention group (21.7 ±2.0,12.8 ± 1.9,17.7 ±2.6) compared with model group (46.8 ±5.3) (all P ＜0.01).Western blot assays demonstrated a significant increase protein level of phosphorylation of p38 MAPK after hypoxia and reoxygenation,and the inhibitor reduced the p-p38 MAPK expression.Compared with the model group,p38 MAPK inhibitor increased the protein and mRNA expression level of Bcl-2,whereas reduced the Bax and caspase-3 expression in the brain.Conclusions Under the influences of p38 MAPK inhibitor,zebrafish larvae improved the behavioral changes after hypoxia-induced brain injury.The inhibitor (10 μmol/L) optimally reduces hypoxia-induced apoptosis in brain by up-regulating Bcl-2,down-regulating Bax/caspase-3 protein and their mRNA level.

BACKGROUND:Because macrophages play an important role in the body’s inflammatory response, the detection of the impact of biological materials on the behavior of macrophages can assess the immunogenicity of materials. OBJECTIVE: To analyze the activation effect of decelularized extracelular matrix materials on macrophages. METHODS: The peritoneal macrophages of BALB/c mouse were obtained and cultured by dividing into five groups. Control group was simple cel culture group, experimental group 1 was acelular matrix membrane material directly contacting with macrophage for culture, experimental group 2 was fresh pericardial material directly contacting with the macrophage for culture, experimental group 3 was acelular matrix membrane material indirectly contacting with macrophages for culture, experimental group 4 was fresh pericardium material indirectly contacting with macrophages for culture. After 24 hours of culture, the secretion of nitric oxide and cytokines in cel culture supernatant was determined. After 48 hours of culture, the absorbance value was determined by MTT method and the toxicity grading was determined. RESULTS AND CONCLUSION: The toxicity grading in experimental groups 1-4 was respectively grades 2, 4, 0, 2. The nitric oxide level in experimental groups 1 and 2 was higher than that in the control group (P < 0.05), and the nitric oxide level in the experimental group 2 was higher than that in the experimental group 1 (P < 0.05). There were no significant differences in interleukin-2, interleukin-4,interferon γ, interleukin-17A and interleukin-10 levels between these five groups. The interleukin-6 level in the experimental group 2 was higher than that in the control group (P < 0.05); The expression levels of tumor necrosis factors in experimental group 1, 2 and 4 were higher than those in the control group (P < 0.05), and experimental group 2 higher than the experimental group 1 (P < 0.05), experimental group 1 higher than the experimental group 4 (P < 0.05). These results show that acelular matrix material can activate macrophages in direct contact.

Objective To verify the indicator reference value ranges of blood cells analytic report by quoting the WS/T 405-2012 industry standard in Changsha area and to establish the reference value ranges of research parameters suitable for local area.Methods The blood cells analytic results in 3138 cases of reference group were analyzed according to the program of the Formulation of Reference Intervals in Clinical Laboratory Detection Items and the results were compared with the WS/T 405-2012 industry standard.Results WBC,Neu#,Mon%,MCHC,PLT,MPV,PCT,P-LCC,P-LCR and InR‰ in the report parameters and WBC-D,PDW-SD in the research parameters had no statistical differences between males and females(P>0.050);WBC,Neu#,Mon%,MCHC and PLT had no difference between genders,the indicators were narrowed compared with the reference value ranges in standard;the reference value ranges of Neu%,RBC,HGB,MCV and MCH in the male blood cells analytic indicators were narrowed,the lower limits of Bas%,Lym#,Mon# and HCT reference value ranges were elevated,the upper limits of Bas%,Lym#,Mon# and HCT reference value ranges were elevated,while the upper limits of Eos# and Eos% reference value ranges were decreased,the Lym% lower limit was decreased;Neu%,Lym#,Mon# and RBC in the female blood cells analytic indicators were narrowed,the lower limits of Bas# and Bas% were increased,the lower limits of Eos%,Lym%,HGB,MCV and MCH were decreased,the upper limits of Eos#,Eos%,Lym%,HGB,MCV and MCH were decreased and the HCT upper limit was elevated.Conclusion The Changsha area should have the reference value ranges of selective quoted standards and establishes the reference value ranges suitable for local area or laboratory.

Objective To assess the vitro susceptibility to 6 antimicrobial agents and genotypes of clinical isolates of Chlamydia trachomatis (Ct) from Guangzhou region. Methods Ct was isolated from clinical specimens by using McCoy cell culture and subjected to propagation. The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of 6 antimicrobial agents (clarithromycin, roxithromycin, azithromycin, doxycycline, tetracycline, ofloxacin) against Ct isolates were determined in McCoy cell culture. Nested PCR was performed to amplify the outer membrane protein 1 (omp1) VS1-2 gene followed by sequencing. Results Seventy-six Ct strains were isolated from 346 urogenital specimens, and 40 strains met the require ments for susceptibility testing after serial propagation. The MIC50/MIC90 of clarithromycin, azithromycin, roxi thromycin, doxycycline, tetracycline and ofloxacin were as follows: 0.008/0.032, 0.080/0.160, 0.125/0.500, 0.032/0.064, 0.250/0.500 and 0.500/1.000 mg/L. Seven genotypes were observed. The most prevalent geno types in decreasing order were E (14, 35%), J (10, 25%)and F (6, 15%). The MIC50 was consistent for azithromycin among the 7 genotypes, but varied by 1 - 4 folds for doxycycline, ofloxacin and roxithromycin. Conclusions Clarithromycin, doxycycline and azithromycin exhibit an excellent activity against Ct, and the activity of azithromycin is consistent among the 7 genotypes of Ct.