Adult ; Electrocoagulation ; adverse effects ; methods ; Fatal Outcome ; Glossopharyngeal Nerve ; surgery ; Glossopharyngeal Nerve Diseases ; surgery ; Humans ; Male ; Microwaves

Objective To investigate the effect of tamoxifen on pituitary tumor transforming gene ex-pression in C6 cell and tumor growth of glioma in vivo. Methods Animal models were established on 32 SD rats with C6 cells of glioma. The rats bearing with C6 glioma were divided into 4 groups randomly, which were treated without tamoxifen or with different doses of tamoxifen(0. 02 mg · kg-1 · d-1, 0. 2 mg · kg-1 · d-1, 2mg · kg-1 · d-1) once a day for 20 days. The dimension of tumors were measured, the tumor inhibition rates were caculated, and living state of the rats were observed. The expression of PTTG mRNA was detected by RTPCR. Results All kinds of doses of tamoxifen could inhibit the tumors growth in rats with C6 glioma, and the tumor volume were reduced by 47.6%, 35.5% and 21.2% in the high-, middle-and low-dose groups respec-tively, there were significantly differences among the 4 groups ( P < 0. 05 ). Low-, middle- and high- dose of tamoxifen all could inhibit the expression of PTTG mRNA, and there were significantly differences among the 4 groups in PTTG mRNA expression ( P < 0. 05 ). Conclusion Tamoxifen can inhibit PTTG expression and the growth of glioma cell line C6 in vivo significantly in dose-dependent manner, which provides a theory basis for clicinal use of tamoxifen in patients with gliomas.

Objective To clone and sequence variant-specific surface antigen gene from Giardia lamblia isolate SUCH/89/BTMRI/2(C2) derived from human in China. Methods Total genomic DNA of G.lamblia was extracted and a full-length variant-specific surface antigen gene fragment was amplified by pelymerase chain reaction (PCR). The PCR product was cloned into pMD19-T simple-vector, transformed into an Escherichia coli JM109 strain and then sequenced. The sequence analysis for cloned fragment was finished by Vector NTI 9.0 software for the homology of Giardia variantspecific surface antigen gene to that of sequences publishend in GenBank. Results The full-length variant-specific surface antigen gone fragment from G. lamblia was found to be 2 142 bp, encoding a 713 amino acid polypeptide and contained a single open reading frame (ORF). The deduced polypeptide sequence was rich in cysteine (11.8 mol%), most of which occurred with in 29 copies of the 4-amino acid CXXC motif, one GGCY-tetrapeptide motifs and three NXS consensus N-linked glycosylation sites. This polypeptide was also rich in threonine (10.2 mol%), glycine (12.1 mol%) and alanine (10.1 mol%). Like other previously identified VSPs, it contained a highly conserved hydropbebic Cterminal region. The homology of G. lamblia SUCH/89/BTMRI/2(C2) variant-specific surface antigen gene to that of sequence (TSA417) published in GenBank was 99% both at the nueleotide and the amino acid levels. Conclusion The full length variant-specific surface antigen gene from the isolate of G. lamblia has the common characteristics with other previously identified VSPs.

OBJECTIVETo observe the effect of Kanli Granule (KG) on myocardial mechanics in pressure overload induced diastolic heart failure (DHF) rats.

METHODSTotally 60 male Wistar rats were divided into the sham-operation group, the model group, the KG group, and the Valsartan group according to random digit table, 15 in each group. The pressure overload induced DHF model was established in all groups except the sham-operation group using abdominal aortic constriction surgery. Totally 7 rats died after modeling (with the mortality of 10. 67%) , and the rest 53 finished the following test. Rats in the KG group were administered with KG extract (calculated as 6. 75 g crude drug/kg) by gastrogavage. Rats in the Valsartan group were administered with Valsartan (7.2 µg/g) by gastrogavage. Equal volume of double distilled water was administered to rats in the model group and the sham-operation group by gastrogavage. All rats were intervened for 32 weeks. The response of isolated heart papillary muscle tonus to isoprenaline (ISO) and adenylate cyclase (Forskolin) was respectively observed. The enhancement phenomenon after resting development force (DF) of isolated heart papillary muscle tonus, and changes of DF in different Ca²⁺ concentrations were observed.

RESULTS(1) In the ISO response test: Compared with the sham-operation group, the amplifications of DF, ±df/dt, -df/dt were obviously elevated in the model group (P < 0.05). Compared with the model group, the amplifications of DF and ±df/dt were obviously lowered in the KG group (P < 0.01), and the amplification of ±df/dt was also reduced in the Valsartan group (P < 0.01). (2) In the Forskolin response test: Compared with the sham-operation group, the amplifications of DF and ±df/dt obviously increased in the model group (P < 0.05). Compared with the model group, the amplifications of DF and ±df/dt were obviously reduced in the KG group (P < 0.01), and the amplification of DF was also reduced in the Valsartan group (P < 0.05). (3) In post-resting DF enhancement test: Compared with the sham-operation group, the amplification of DF showed gradually decreasing tendency along with prolonged resting time in the model group, and they were obviously lowered at all time points (P < 0.05). Compared with the model group, the amplification of DF was gradually increasing along with prolonged resting time in the KG group. The amplification of DF at post-resting 240 s was obviously larger in the KG group than in the model group (P < 0.05). The amplification of post-resting DF still showed gradually decreasing tendency along with prolonged resting time in the Valsartan group, with increased amplifications of DF at post-resting 60 s and 120 s (P < 0. 05) (4) The amplifications of DF in different Ca²⁺ concentrations: Compared with the sham-operation group, the amplifications of DF were significantly elevated in different Ca²⁺ concentrations (1.75, 3.5, 7.0 mmol/L ) (P < 0.05, P < 0.01). Compared with the model group, there was no statistical difference in amplification of DF in different Ca²⁺ concentrations in the KG group (P > 0.05). The amplifications of DF in different Ca²⁺ concentrations were significantly reduced in the Valsartan group (P < 0.05).

CONCLUSIONSThe ISO response and the Forskolin response were enhanced in isolated heart papillary muscle tonus of pressure overload induced DHF rats; enhanced post-resting DF was reduced; DF in different supra-physiologic levels of Ca²⁺ was still enhanced. KG could significantly improve excessive enhancement of pressure overload induced DHF rats in ISO response and Forskolin response, and improve enhancement of post-resting myocardium.

Animals ; Colforsin ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Heart ; drug effects ; physiopathology ; Heart Failure, Diastolic ; drug therapy ; Isoproterenol ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Wistar

Background C57BL/6(B6) is a kind of routine mouse specie used in experimental autoimmune uveitis (EAU) research.Previous studies showed that the pathogenesis of uveitis related to inflammatory cytokines secreted by different helper T(Th) cells.However,the interaction of different Th cells in EAU is unclear.Objective This study was to investigate the dynamic changes of inflammatory factors in the spleen and serum after immunization in EAU mice.Methods Forty-four SPF B6 mice were immunized by injection of interphotoreceptor retinoid-binding protein (IRBP) and complete Freund adjuvant (CFA) emulsion via caudal vein and footpad.Indirect ophthalmoscope was used to examine the eyes 3 times per week and the inflammatory response was scored based on Thurau's criteria.In the thirty day after injection,20 model eyes were extracted and the sections of eye tissue were prepared for histopathological examination.The spleens of model mice were enucleated before injection and 2,5,10,15,20,25,30 days after injection,and reverse transcriptase PCR (RT-PCR) was used to detect the contents of interleukin-17 (IL-17) mRNA,interferon-γ (IFN-γ) mRNA,tumor necrosis factor-α (TNF-α) mRNA and IL-10 mRNA,and the contents of IL-17,IFN-γ,TNF-α and IL-10 in model serum were assayed by ELISA in 24 model mice.The experimental protocol and use of the animals were approved by Ethic Committee for Care and Use of Laboratory Animals of Shandong University of Traditional Chinese Medicine.Results Mild inflammatory response was seen in 12 days under the indirect ophthalmoscope with the scores of 0.5.The inflammatory scores peaked in 13-15 days with the scores of 1.0 and alleviated after that with the inflammatory scores of 0.5 in 30 days after injection.The histopathological score was consistent with the clinical score in the models on the 30 days.The serum IL-17 content of model mice was (0.98±0.05) ng/L before injection and increased to (51.85 ±2.42) ng/L on the fifth day,and decreased to (4.01±0.06)ng/L on the fifteen day.But,the serum IL-17 level increased to (25.00±0.94)ng/L again on the 25th day,and then lowed to (6.01 ±0.21)ng/L 30 days after injection,showing a significant elevation in comparison with that of before injection (P=0.000).The serum IFN-γ content of the model mice was (1.02±0.09)ng/L before injection and increased to (50.54±0.48) ng/L on the fifth day,and (73.21±0.12) ng/L on the tenth day,and then it declined gradually until (5.15±0.18)ng/L in the 30th day,which was still higher than that of before injection (P=0.000).After injection of IRBP+CFA,the serum TNF-α level upregulated from the second day to fifth day with the peak values (134.25±0.59)ng/L,and declined to valley on 15th day.A repeat elevation of serum TNF-α level was found on the 20th day with the values (60.54±0.62)ng/L and followed by decrease till the 30th day,which was higher than that of before injection (P=0.660).Serum IL-10 was detectable in the tenth day and peaked on the 15th day.Then a slight decrease was seen till the 30th day,compared with before injection(P =0.000).The contents of IL-10 mRNA,IL-17 mRNA,TNF-α mRNA,IFN-γmRNA in mice spleens followed the same pattern with serum levels of their proteins.Conclusions IL-17,IFN-γ,TNF-α and IL-10 are key inflammatory factors of Th1,Th2 and Th17,they present with specific changes during EAU,it confirming that IFN-γ probably play a pathogenic role in EAU,IL-17 and TNF-α levels probably associated with the chronic and recurrent procedure of uveitis,IL-10 plays an inhibit role in EAU.

Background Lewis rat is a commonly used specie in experimental autoimmune uveitis(EAU).However,the characteristics of EAU,especially ocular uhrastructural change,are rarely reported.Objective This study was to investigate the inflammatory characteristics and ocular uhrastructure of EAU models in Lewis rat.Methods EAU models were induced in 12 SPF female Lewis rats(6-8 weeks old)by injection of complete Freund adjuvant(CFA) containing interphotoreceptor retinoid-binding protein (IRBP,1177-1191) and tuberculin (TB) into footpads,napes and back of the body,and 6 normal matched rats were used as normal controls.The diet and drinking,temperature and behavior acts of the rats were recorded during the observational duration.Ocular manifestations were examined under the slit lamp biomicroscope after modeling and scored.Eyeballs were obtained in 12 days after modeling for histopathological examination,and the ultrastructures of eyeballs were observed under the scanning electron microscope and transmission electron microscope.The use and care of the animals complied with Statement of ARVO.Results The food-intake was (190.00± 18.03)g in the model group,and that in the control group was (285.33 ±28.02) g,showing a significant difference between them (t =4.955,P =0.012).The drinking-water volume of rats was (241.67±t 18.56)ml in the model group,which was significantly less than (289.67± 18.18)ml in the control group,showing a significant difference between them (t =3.201,P =0.033).In addition,elevated temperature and tiredness were found in the model rats.Anterior chamber empyema,iris hyperemia and occlusion of the pupil appeared in the models on the 6th day and peaked on the 12nd day after immunized,with the inflammatory scores of 3.83±0.41.The infiltration of inflammatory cells were seen in anterior chamber,ciliary body and vitreous cavity under the optical microscope in the model rats.Scanning electron microscopy found uneven iris texture,coarse ciliary surface and loosen villi of retinal pigment epithelial (RPE) cells in the model rats.Under the transmission electron microscope,the infiltration of macrophagocytes on the iris,wrinkle sparse of ciliary epithelium,myeloid bodies in retinal Müller cells and vacuolus in mitochondria of RPE cells were exhibited.No obvious abnormality was found in the control rats.Conclusions Lewis rats are autoimmune status following injection of CFA with IRBP(1177-1191) and TB.The morphology and ultrastructure of eyeballs in the EAU rats can explain the finding of eyes.

Objective To examine the relationship between CCT 5 ,γδ T cell and autoimmune diseases .Methods Recombinant CCT5 protein was cloned , expressed and purified in E.coli.Three peptides of CCT5 protein were used to prepare for anti-CCT5 monoclonal antibodies .Purified CCT5 protein was used to expand γδT cells from pe-ripheral blood mononuclear cells (PBMC).Plasma level of CCT5 in healthy donors, patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) were detected by ELISA assays .The correlation analysis between plasma CCT5 concentration and the percentage of different subtypes of γδT cells measured by flow cytometry was made . Results The CCT5 gene was amplified by PCR and the length of the target fragment was 1 750 bp.The expressed 65 ku CCT5 protein was purified and validated by SDS-PAGE.Two paired monoclonal anti-CCT5 antibodies were screened to detect CCT5 protein in plasma.Immobilized recombinant CCT5 protein was able to induce specific sig-nificant amplification of peripheral γδT cells.Correlation analysis of 10 healthy donors indicated significant corre-lation between the plasma CCT 5 concentration and the proportion of Vγ9 and Vδ2 γδ T cells.The plasma CCT5 concentration significantly decreased in autoimmune diseases patients , including RA and SLE .Conclusions These data suggest that CCT 5 could be a novel Vγ9δ2 γδT cell-related factor in autoimmune diseases , which deepen the understanding of Vγ9δ2 γδT cell function in autoimmune diseases .

Objective To study the influence of Annao tablet on the expression of transforming growth factor beta 1 (TGF-β1) in acute graft-versus-host disease (aGVHD) murine and to explore the interventional mechanism of TGF-β1 on aGVHD. Methods Hematopoietic stem cells of male Balb/c mice were transplanted to female C57BL/6 mice for the development of aGVHD murine model. Recipient mice were divided into Annao group and blank group randomly and respectively administrated with Annao soup (a kind of Chinese herb) and 0.9% sodium chloride intragastrically. Clinical symptom, survival time and body weight were recorded at 14th and 30th day and some sections of liver, small bowel and skin were taken for histological changes. Serum level of TGF-β1 were measured by enzyme-labeled immunosorbent assay (ELISA), splenocyte protein of TGF-β1 by Western Blot and TGF-β1 mRNA by fluorescent quantitation polymerase chain reaction (PCR). Results Serum level of TGF-β1 in both groups had no statistical difference (P = 0.305), but it rose to (148.31 ± 7.95) ng/mL at 14th day and (183.48 ± 5.91) ng/mL at 30th day in Annao group, which had significant difference when compared with that in blank group (P = 0.000). IOD/IODβ-actin value of TGF-β1 protein in Annao group was 0.33 ± 0.05 at 14th day and 0.56 ± 0.04 at 30th day, which was higher than that in blank group (P = 0.000) and the expression of TGF-β1 mRNA of splenocyte in Annao group was 1.24 ± 0.04 at 14th day and 2.14 ± 0.33 at 30th day which was much higher than that in blank group (P = 0.000). Conclusion Annao tablet helps to relieve symptoms of acute GVHD by raising serum level of TGF-β1 and intensifying expression of protein of TGF-β1 and its mRNA.

BACKGROUND: At present, the materials that are successful y applied in rib fracture internal fixation mainly include titanium, nitinol rib encircling bone bonding plate, imported poly-L-lactic acid absorbable rib nails and anatomical plate.
OBJECTIVE: To study the biomechanical characteristics of new magnesium al oy absorbable rib intramedul ary nail in the rib fracture fixation, and to compare with imported poly-L-lactic acid absorbable rib intramedul ary nail in order to provide scientific basis for clinical application.
METHODS: Thirty fresh adult fifth rib specimens were col ected, and the specimens were used to make models for the middle rib fractures. The specimens were fixed with AZ31B magnesium al oy absorbable rib intramedul ary nail (magnesium al oy group) and poly-L-lactic acid absorbable rib intramedul ary nail (poly-L-lactic acid group), and the normal rib specimen group was set as control. The biomechanical characteristics of the nails in each group were tested with stress analysis.
RESULTS AND CONCLUSION: Three-point bending strength measurement results showed that the bending strength in the magnesium alloy group was close to the normal value of the specimen (P > 0.05), and there was significant difference in the bending strength between magnesium alloy group and the poly-L-lactic acid group (P < 0.05). Torsional strength measurement showed that there was no significant difference in torsional strength between magnesium alloy group and the normal specimens, and the results showed that the magnesium alloy was better than poly-L-lactic acid in fixation (P < 0.05). Tensile tests showed that the tensile strength and anti-pulling force of the magnesium alloy fixation were better than those of poly-L-lactic acid fixation (P < 0.05). The results indicate that the magnesium alloy absorbable rib intramedullary nail is better than poly-L-lactic acid absorbable rib intramedullary nail in strength and tensile strength, which is the ideal fixation material for rib fixation.

Objective To compare the outcomes of traumatic flail chest with multiple injuries treated by operative fixation versus conservative approach. Methods The clinical data of 60 patients with traumatic flail chest with multiple injuries were retrospectively analysed, and the outcomes between operation group (treated by operative fixation, n=32)and non-operation group (treated by conservative approach, n=28) were compared. Results The mean time of hospital stay, ICU stay and mechanical ventilation was significantly shorter, and the prevalences of chest wall deformity, pulmonary infection, pulmonary atelectasis and respiratory failure were significantly lower in operation group than those in non-operation group (P<0.05). Six months after discharge, the pulmonary function parameters such as inspiratory capacity, forced vital capacity, forced expiratory volume in one second, peak expiratory flow, total lung capacity and maximal midexpiratory flow were significantly higher in operation group than those in non-operation group (P<0.05). Conclusion Traumatic flail chest with multiple injuries treated by operative fixation may lead to less flail chest associated complications. Operative fixation has short- and long-term benefits to flail chest.