Preparation and immunogenicity evaluation of recombinant influenza hemagglutinin trimer vaccine / 中华微生物学和免疫学杂志

Objective:To prepare a recombinant hemagglutinin trimer (HA-Tri) vaccine against influenza viruses and to study its immunogenicity in a mouse model.Methods:A stable CHO cell line that could express HA-Tri was constructed. Western blot, single radial immunodiffusion, protein particle size detection and N-glycosylation site analysis were performed for qualitative and quantitative analysis of the recombinant protein. According to the different treatment conditions such as dosage and adjuvant, BALB/c mice were divided into 11 groups and subjected to consistent immunization procedures. Serum neutralizing antibody titers were measured on 56 d after the first immunization to evaluate the immunogenicity of HA-Tri.Results:The constructed CHO cells could secret and express HA-Tri proteins. The HA-Tri proteins were biologically active and capable of forming precipitation rings in the single radial immunodiffusion. The particle size of HA-Tri was approximately 18.79 nm and 10 N-glycosylation sites were detected, including high mannose, complex glycoforms and heterozygous glycoforms. After prime-boost immunization, there was no statistically significant difference in the titers of neutralizing antibodies induced in mice by 3.75 μg of HA-Tri in combination with RFH01 adjuvant and 15 μg of monovalent vaccine stock solution ( P=0.431 2, U=36). Serum antibody titers in the HA-Tri+ RFH01 groups were higher than those in the corresponding HA-Tri groups without RFH01 adjuvant, and the highest titer was induced in the 15 μg HA-Tri+ RFH01 group, which was 1 280. Conclusions:The recombinant HA-Tri protein was successfully prepared. HA-Tri in combination with RFH01 adjuvant could induce humoral immune responses against influenza viruses in BALB/c mice, which would provide reference for the development of influenza virus recombinant subunit vaccines.

Advances of Researchs on Molecular Mechanisms of Mesenchymal Stem Cells and Their Exosomes in Angiogenesis--Review / 中国实验血液学杂志

Mesenchymal stem cells (MSC) have the potential of multi-directional differentiation, and can recruit endothelial cells, promote their proliferation, migration and angiogenesis, improve blood perfusion and oxygen suppliment, and repair damaged tissue. Exosome secreted by MSC contain mother cell-specific proteins, lipids and nucleic acids, and acts as signaling molecule, playing an important role in cell communication, thereby altering target cell function. In this review, the biological characteristics of MSC and its exosome, the mechanism of promoting vascular regeneration in patients with ischemic diseases, and the mechanism of hypoxia-inducible factor-1α（HIF-1α） in the vascular ischemia of ischemic diseases are all summarized briefly.

The risk factors for predicting or diagnosing bone metastases from urinary bladder neoplasms / 解放军医学杂志

Objective To explore the clinical significance of some risk factor,such as blood calcium serum alkaline phosphatase (ALP) and hemoglobin (Hb) for predicting or diagnosing bone metastases from urinary bladder neoplasms.Methods A total of 35 patients diagnosed as urinary bladder neoplasms and bone metastasis based on the histopathologic analysis (bone metastasis group,BM),and 105 cases of age-,sex-and follow-up time matched urinary bladder neoplasms patients without bone metastasis (non bone metastasis group,NBM) were included in present study.The correlations between clinical-pathological parameters (blood calcium,ALP,Hb) and bone metastases from urinary bladder neoplasms were analyzed and the risk factors for predicting or diagnosing bone metastases were identified.Results The levels of blood calcium,ALP and Hb were significantly higher at first diagnosis and follow-up period in BM group than in NBM group (P＜0.05).Multivariate logistic regression analysis indicated that the levels of calcium,ALP and Hb at first diagnosis were the independent risk factors for predicting the later incidence of bone metastases (P＜0.05).The levels of calcium,ALP and Hb in follow-up period were the independent risk factors for diagnosing bone metastases (P＜0.05).Receiver operating characteristic (ROC) curves analysis showed that the levels of calcium,ALP,Hb may give a more accurate prediction on bone metastases (AUC:0.615,0.704,0.692;cut-off:2.55mmol/L,97.0U/L,101.5g/L,respectively) or diagnosing bone metastases (AUC:0.706,0.776,0.829;cut-off:2.46mmol/L,112.5U/L,103.5g/L,respectively).Conclusion The levels of blood calcium,ALP and Hb may be used to predict the risk of bone metastases for patients firstly diagnosed as urinary bladder neoplasms,and predict whether the patients will be complicated with bone metastases during follow-up period.

Effects of sleeve gastrectomy in mice by using titanium clip / 国际外科学杂志

Objective To introduce details of a new procedure of sleeve gastrectomy in mice by using titanium clip,and to observe whether this model has the same post-surgical metabolic effects as traditional procedure of sleeve gastrectomy in mice by using running stitch.Methods Thirt-one male FVB-Tg (NPY-hrGFP) mice,6 weeks (29-31 g) were randomly divided into SG (running stitch) group (n =8),SG (titanium clip) group (n =15) and Sham Operation (SO) group (n =8) by using random number table.Titanium clip was applied to close the side incision of stomachin SG (titanium clip) group,while running stitch was used in SG (running stitch) group which was detailed described in current references.Ligaments around stomach were detached without any stomach incision in SO group.Operation duration,mortality,and changes of post-surgical body weight and food intake were observed.Measured data were expressed as mean ± standard deviation ((x) ± s).Independent sample t test was used for comparison between the two groups.The variance of the data between the three groups was equal and analysis of variance (least significant LSD method) was used to compare each group.Preoperative and postoperative data were compared using a paired t test.Results Operation duration of SG (running stitch) group and SG (titanium clip) group were (44.60 ± 3.13) min and (28.25 ± 2.77) min,respectively.Compared with SG (running stitch) group,operation duration was significantly reduced in SG (titanium clip) group (P ≤ 0.01).Mortality of SG (running stitch) group and SG (titanium clip) group were 37.5％ and 20％,respectively,which has no significant difference (P =0.621).Compared with SO group,postoperative food intake was significantly reduced in both two SG groups (P ≤ 0.01),but there's no difference between two SG groups (P =0.149).At the end of the 6-week-postoperative,body weights of SG (running stitch) group and SG (titanium clip) group were reduced (17.67 ± 5.06) ％ and (18.76 ± 7.87) ％,respectively,while body weight of SO group increased (7.17 ± 2.31)％.Compared with SO group,postoperative change of body weight was significantly reduced in both two SG groups (P≤ 0.0 1),but there's no difference between two SG groups (P =0.746).Conclusions SG by using titanium clip in mice can reduce food intake and body weight as traditional SG by using running stitch does.But with shorter operation duration,this new model of SG in mice can provide a better and stable surgical model in research of mechanism of SG.

The role of microRNAs in bone remodeling / 国际口腔科学杂志·英文版

Bone remodeling is balanced by bone formation and bone resorption as well as by alterations in the quantities and functions of seed cells, leading to either the maintenance or deterioration of bone status. The existing evidence indicates that microRNAs (miRNAs), known as a family of short non-coding RNAs, are the key post-transcriptional repressors of gene expression, and growing numbers of novel miRNAs have been verified to play vital roles in the regulation of osteogenesis, osteoclastogenesis, and adipogenesis, revealing how they interact with signaling molecules to control these processes. This review summarizes the current knowledge of the roles of miRNAs in regulating bone remodeling as well as novel applications for miRNAs in biomaterials for therapeutic purposes.

Pilot study of the damage degree of Henoch-Schonlein purpura nephritis on children with acoustic radiation force impulse imaging / 中华超声影像学杂志

Objective To assess the value of acoustic radiation force impulse (ARFI) imaging in the pathological damage of Henoch-Schonlein purpura nephritis on children by comparison with renal biopsy.Methods 50 cases of healthy control group as group A; 58 children with HSPN were divided into three groups according to pathological grading:Ⅰ ～ Ⅱ class (group B),Ⅲ class (group C) and Ⅳ ～ Ⅵ class (group D).ARFI was then used to measure the shear wave velocities(SWV) of renal cortex of each group,compared the differences SWV of each group.Results SWV values of children's renal cortex with HSPN were significantly higher (t =5.883,P =0.017) than those in the group A.Pairwise comparisons found that there were statistically significant differences between group D and the other three groups (P ＜0.05).According the ROC curve,the cut-off value of SWV was 2.59 m/s when the maximum area under the curve equal to 0.719,the sensitivity and specificity were 63％ and 67％.Conclusions ARFI technology can quantify the elastic properties of the kidney,which is expected as an important indicator to evaluate the pathological extent of damage of the HSPN.

Analysis of X-rays of cervical imbalances syndrome before and after the treatment of osteopathy and traction intervention in 187 youth cases / 中国骨伤

<p><b>OBJECTIVE</b>To observe X-ray features of before and after treatment of cervical imbalance syndrome with osteopathy and traction intervention in youth patients, then to investigate the clinical effect of the treatment of the cervical imbalances syndrome in youth by osteopathy.</p><p><b>METHODS</b>From September 2007 to December 2010, one hundred and eighty-seven students of 19 to 22 years (means 21 years) with neck pain as the main symptom in Beijing university of Chinese medicine were selected and divide them into osteopathy group and traction groups randomly. In osteopathy group, there were 94 patients including 40 males and 54 females; in traction group, there were 93 patients including 42 males and 51 females. The treatment of osteopathy group lasted for 3 weeks,three times per week; traction group by traction treatment three weeks,three times a week, too. X-ray before treatment and 3 weeks after treatment were collected.</p><p><b>RESULTS</b>The osteopathy group:94 cases, before treatment, abnormal curvature in 57 cases, spinous position change in 45 cases and angular displacement in 44 cases, vertebral sliding in 15 cases, Ruth Jackson line intersect proneness change in 70 cases,stretch change in 47 cases; after treatment, abnormal curvature in 35 cases, spinous position change in 24 cases and angular displacement in 18 cases, vertebral sliding in 3 cases, Ruth Jackson line intersect proneness change in 41 cases, extension change in 33 cases; X-Ray measurement results: cervical curvature improved from (7.070 +/- 4.629) degrees before treatment to (7.660 +/- 4.156) degrees after treatment, angular displacement improved from (13.790 +/- 2.590) degrees before treatment to (11.050 +/- 2.560) degrees after treatment; vertebral sliding improved from (3.770 +/- 0.350) mm before treatment to (3.160 +/- 0.485) mm after treatment. The change of angular displacement and vertebral sliding were so significant before and after treatment in this group, there were statistical defferences between before and after the treatment (P < 0.01) in osteopathy group. The traction group: 93 cases, before treatment,abnormal curvature in 60 cases, spinous position change in 39 cases and angular displacement in 39 cases, vertebral sliding in 15 cases, Ruth Jackson line intersect proneness change in 70 cases, stretch in 47 cases; after treatment,abnormal curvature in 50 cases, spinous position change in 29 cases and angular displacement in 17 cases; vertebral sliding in 3 cases, Ruth Jackson line intersect proneness change in 41 cases, stretch in 33 cases; X-Ray measurement results: cervical curvature improved from (5.590 +/- 4.639) degrees before treatment to (5.990 +/- 4.330) degrees after treatment, angular displacement improved from (13.360 +/- 2.064) degrees before treatment to (11.210 +/- 1.872) degrees after treatment; vertebral sliding improved from (3.790 +/- 0.339) mm before treatment to (3.480 +/- 0.332) mm after treatment. The change of angular displacement and vertebral sliding were also so significant in traction group before and after treatment, there were statistical defferences between before and after the treatment (P < 0.01).</p><p><b>CONCLUSION</b>(1) Pathological changes can be called youth cervical imbalance syndrome. (2) Stress points, angular displacement, cervical vertebral slip, curvature straightened, spinous position change are main X-ray performances. (3) Both osteopathy and traction intervention on are efficient in youth neck pain. (4) Youths cervical vertebra imbalance is early but reversible performance.</p>

Palmitate induces apoptosis and endoplasmic reticulum stress in human umbilical cord-derived mesenchymal stem cells / 生理学报

The saturated free fatty acid (FFA), palmitate, could induce apoptosis in various cell types, but little is known about its effects on human umbilical cord-derived mesenchymal stem cells (hUC-MSCs). Here, we investigated whether palmitate induced apoptosis and endoplasmic reticulum (ER) stress in hUC-MSCs. hUC-MSCs were stained by labeled antibodies and identified by flow cytometry. After administration with palmitate, apoptotic cell was assessed by flow cytometry using the Annexin V-FITC/7-AAD apoptosis detection kit. Relative spliced XBP1 levels were analyzed using semi-quantitative RT-PCR. The mRNA of BiP, GRP94, ATF4 and CHOP were analyzed by real-time PCR. Relative BiP and CHOP protein were analyzed using Western blot analysis. The results showed that hUC-MSCs were homogeneously positive for MSC markers; palmitate increased apoptosis of hUC-MSCs and activated XBP1 splicing, BiP, GRP94, ATF4 and CHOP transcription. These findings suggest that palmitate induces apoptosis and ER stress in hUC-MSCs.

Hepatitis B e antigen from chronic hepatitis B patients induces Th1/Th2 cytokine imbalance in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the immunoregulatory effect of hepatitis B virus (HBV) e antigen (HBeAg) on peripheral blood monocytes (PBMCs).</p><p><b>METHODS</b>PBMCs were isolated from patients with chronic hepatitis B (CHB; both HBeAg- and HBeAg+) and healthy controls, and cultured with recombinant HBeAg. The HBeAg-induced changes in expression of PD-1/PD-L1 were measured by flow cytometry of the cells and in secreted cytokines were measured by enzyme-linked immunosorbent assay of the supernatants. Comparisons between two groups were made by the independent-samples t-test; the relationship between PD-1/B7-H1 level and HBV DNA copy number was evaluated by Spearman's correlation analysis.</p><p><b>RESULTS</b>Exposure to HBeAg led to a significant decrease in CD3+CD4+ T lymphocyte-specific expression of IFNa for both the CHB patients' and healthy controls' samples (t = 2.382 and -4.190 respectively, P less than 0.01). For the HBeAg- CHB patients' and healthy controls' samples, the HBeAg exposure led to increased levels of secreted cytokines IL-6, IL-10 and TNFa (t = 2.504, 3.583 and 4.324, P less than 0.01 and t = 3.542, 6.246 and 5.273, P less than 0.01 respectively) and of CD14+ PBMC-specific expression of PD-L1 (t = 4.815 and 3.454, P less than 0.05 respectively). Compared to the HBeAg-negative CHB patients' and healthy controls' samples, the HBeAg+ CHB patients' samples had significantly lower CD3+CD4+ T cell-specific expression of IFNa (t = -3.177 and -4.541, P less than 0.01 respectively), but significantly higher levels of secreted IL-4 (t = 3.382 and 4.393, P less than 0.01 respectively), of CD3+ T cells-specific expression of PD-1/PD-L1 (t = 4.755, 2.942 and 4.518, 4.595, P less than 0.01 respectively), and of CD14+ T cells-specific expression of PD-L1 (t = 5.092 and 5.473, P less than 0.01 respectively). The CD3+ T cells-specific expression of PD-L1 was significantly higher in the samples from HBeAg- CHB patients than from the healthy controls (t = 3.214, P less than 0.01).</p><p><b>CONCLUSION</b>HBeAg was able to down-regulate the production of Th1-type cytokines (IFNgamma), and up-regulate the secretion of Th2-type cytokines (IL-6, IL-10) and the expression of PD-1/PD-L1on monocytes. These changes are conducive to the formation of immune tolerance to HBV. Therefore, HBeAg may play an important role in immune tolerance to chronic HBV infection.</p>

Value of ultrasonography in the diagnosis of infantile hepatic hemangioendothelioma / 中华超声影像学杂志

Objective To investigate the ultrasonographic performance of infantile hepatic hemangioendothelioma(IHHE).Methods Clinical and ultrasonographic findings in 14 cases with IHHE were reviewed retrospectively,and compared with CT and MR.Results Among 14 cases,there were 6 cases with single mass,5 cases with multiple masses and 3 cases with diffuse masses.Ultrasonography displayed single mass and multiple masses with clear boundary,heterogeneous inner echo,calcification was detected in 4 cases and anechoic areas were detected in 6 cases.Diffuse masses were small,ill-defined,the echogenicity of the whole liver was extremely disorder.The hepatic artery and the proximal abdominal aortic dilatation was found in 4 cases,and the abdominal aorta which below the coeliac artery became relatively thinner in these cases.Color Doppler showed rich blood supply and intratumoral vasodilation in 8 cases,PW detection of intratumoral arteriovenous fistula in 4 cases.Conclusions Ultrasonography findings of IHHE have some characteristic feature.

Multiple linear regression analysis of X-ray measurement and WOMAC scores of knee osteoarthritis / 中国骨伤

<p><b>OBJECTIVE</b>To perform Multiple Linear Regression analysis of X-ray measurement and WOMAC scores of knee osteoarthritis, and to analyze their relationship with clinical and biomechanical concepts.</p><p><b>METHODS</b>From March 2011 to July 2011, 140 patients (250 knees) were reviewed, including 132 knees in the left and 118 knees in the right; ranging in age from 40 to 71 years, with an average of 54.68 years. The MB-RULER measurement software was applied to measure femoral angle, tibial angle, femorotibial angle, joint gap angle from antero-posterir and lateral position of X-rays. The WOMAC scores were also collected. Then multiple regression equations was applied for the linear regression analysis of correlation between the X-ray measurement and WOMAC scores.</p><p><b>RESULTS</b>There was statistical significance in the regression equation of AP X-rays value and WOMAC scores (P<0.05), while there was no statistical significance in the regression equation of lateral X-ray value and WOMAC scores (P>0.05).</p><p><b>CONCLUSION</b>1) X-ray measurement of knee joint can reflect the WOMAC scores to a certain extent. 2) It is necessary to measure the X-ray mechanical axis of knee, which is important for diagnosis and treatment of osteoarthritis. 3) The correlation between tibial angle,joint gap angle on antero-posterior X-ray and WOMAC scores is significant, which can be used to assess the functional recovery of patients before and after treatment.</p>

The clinical application of infrared imaging technology in teenagers with cervicodynia / 中国骨伤

<p><b>OBJECTIVE</b>To compare the temperature distribution of both sides of shoulder, provide objective reference for the application thermoview in the diagnosis of cervicodynia in teenagers.</p><p><b>METHODS</b>Forty-five adolescents with cervicodynias from March 2009 to December 2009 were collected. There were 23 males and 22 females, with an average age of 21 years (ranged from 19 to 22 years). The course of disease ranged from 2 to 20 weeks (averaged 13 weeks). C7 horizontal line were used to divide the back into the neck area and shoulders area, and the midline to subdivide the chosen area into left and right area. Thermal infrared imaging was used to observe the temperature both sides of neck and shoulders, the data was analyzed by the computer.</p><p><b>RESULTS</b>The temperature of shoulder was higher than neck. There were significant differences in the highest and average temperature both of the left and right side of neck and shoulder (P < 0.05), but no difference in the lowest temperature both of left and right side of neck.</p><p><b>CONCLUSION</b>The thermal infrared imaging is important for diagnosing cervical imbalance syndrome, finding the sign of abnormal muscle metabolism of shoulder and providing the basis for prevention.</p>

Case-control study on therapeutic effects between bone-setting and herbal fumigation for the treatment of the obsolete malleolus joint sprains / 中国骨伤

<p><b>OBJECTIVE</b>To compare the clinical effects of bone-setting technique and herbal fumigation for the treatment of the obsolete malleolus joint sprains.</p><p><b>METHODS</b>From March 2008 to May 2011, 76 patients were divided into treatment group (39 cases) and control group (37 cases). In the treatment group: 15 males and 24 females; the age ranged from 20 to 59 years with an average of (42.97 +/- 9.21) years; the course of disease ranged from 1 to 60 months; the average score of ankle joint function was (71.27 +/- 4.50). In the control group: 11 males and 26 females; the age ranged from 25 to 57 years with an average of (41.29 +/- 8.77) years; the course of disease ranged from 1 to 36 months with an average of (8.47 +/- 7.37) months; the average score of ankle joint function was (71.45 +/- 4.61). The patients in the treatment group were treated with bone-setting technique two times a week, and the patients in the control group were treated with herbal fumigation once a day. The ankle joint function scores and treatment effects of the two groups were compared after 3 weeks by using Baird-Jackson ankle function score.</p><p><b>RESULTS</b>After 3 weeks of the treatment, the average score of ankle joint function of the treatment group was (93.44 +/- 4.91), and in the control group was (85.8 +/- 16.57), the difference has statistical significance. The treatment group score was better than that of the control group. Before and after treatment,the average ankle score of the treatment group was (71.27 +/- 4.50) and (93.44 +/- 4.91), the difference has statistical significance. Before and after treatment, the average ankle score of the control group was (71.45 +/- 4.61) and (85.81 +/- 6.57), the difference has statistical significance. In the treatment group, 16 cases got an excellent result, 18 good, 3 fair, 2 poor; in the control group, 9 cases got an excellent result, 14 good, 5 fair,9 poor. The difference has statistical significance.</p><p><b>CONCLUSION</b>The bone-setting techniques and herbal fumigation treatment of obsolete malleolus joint sprains both have a certain effect, and the former is better than the latter.</p>

Dynamic changes and clinical significance of HBcAg18-27 specific cytotoxic T lymphocytes in acute hepatitis B patients / 中华肝脏病杂志

This report aims to investigate the dynamical changes of HBcAg18-27 epitope specific cytotoxic T lymphocytes(CTL), alanine aminotransferase (ALT), HBV DNA and HBsAg in peripheral blood of acute hepatitis B patients, and to explore the roles of HBcAg18-27-specific CTLs in virus clearance and liver injury. Acute hepatitis B (AHB) and chronic hepatitis B (CHB) patients were divided into two groups according to results of HLA-A0201. Patients with positive HLA-A0201 were classified into HBcAg-specific CTL group and those with negative HLA-A0201 were referred as control group. The specific CTLs were stained with HLA-A0201 limited HBcAg18-27 epitope MHC-Pentamer and the frequencies of CTLs, T, B, NK and NKT cells were detected by flow cytometry (FCM). The serum ALT, HBV DNA and HBsAg were examined using speed analysis, quantitative PCR and abbott chemiluminescent technology. The frequencies of HBcAg18-27-specific CTLs in AHB patients were higher in the early three weeks as compared to the late three weeks. The apex time of HBV-specific CTL frequencies lagged behind those of HBV DNA, HBsAg and ALT. The loss of HBsAg in patients with high frequencies of HBV-specific CTL was earlier than that in patients with low frequencies (t = 2.018, P value is less than 0.05). In the second week the peak frequencies of CD3+CD8+ cells overlapped with that of HBcAg18-27-specific CTLs and with a positive correlation between (r = 0.420, P value is less than 0.05). During the early stages of AHB, the frequencies of NK and NKT cells were found significantly lower than that of control group and CHB group and the levels were back to normal after recovery. Moreover, a negative correlation existed between the frequencies of NK cells and the dynamic changes of HBcAg18-27-specific CTLs (r = -0.435, P value is less than 0.01) in AHB group. The frequencies of HBcAg18-27-specific CTLs were significantly higher as compared to CHB group in the first three weeks (z = -3.258, -4.04, and -3.259, P value is less than 0.01). The early loss of HBsAg was closely related to the high frequencies of HBcAg18-27 specific CTLs in AHB patients. HBcAg-specific CTL frequencies in peripheral blood could be used to predict clinical outcome after HBV infection. The frequencies of CD8+ T cells can reflect the changes of frequencies of HBcAg-specific CTL during acute HBV infection.

A novel missense mutation resulting in X-linked adrenoleukodystrophy in female heterozygotes of a Chinese family / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify ABCD1 gene mutation in a Chinese family with three heterozygous female patients.</p><p><b>METHODS</b>Four fragments covering the entire coding sequence of the ABCD1 gene from one of the female patients were amplified by reverse transcription-PCR. The PCR products were directly sequenced. The result of sequencing was confirmed by restriction enzyme digestion of PCR products from genomic DNA. Human ABCD1 gene and ALD protein were aligned with those of rat, monkey, mouse and cattle by Clustal X 1.83. Softwares of Motif Scan, TMpred and ESYpred3D were used to predict the effect of the mutation on the structure of the ALD protein.</p><p><b>RESULTS</b>A novel missense mutation, CAC to CGC, was found at codon 283 of the ABCD1 gene from the patient, resulting in the replacement of histidine by arginine. This mutation abolished an Msl I site in the gene. Her son was free from this mutation. The mutated amino acid residue (283H) was highly conservative in evolution, and the mutation caused a dramatic change in the structure of the ALD protein.</p><p><b>CONCLUSION</b>Three female patients heterozygous for ABCD1 gene mutation were first reported in China, and a novel mutation, p.H283R, was identified in this X-ALD family.</p>

Down-regulation of mTOR activity and survivin expression during tamoxifen-induced apoptosis in hepatoblastoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to investigate the changes in mTOR activity and survivin expression in liver cancer cell line HepG2 cells treated with tamoxifen.</p><p><b>METHODS</b>Survivin transcription level and p70S6K was demonstrated by PCR, dual-luciferase reporter assay and Western blot analysis, respectively, and the apoptosis in the HepG2 cells was detected by flow cytometry.</p><p><b>RESULTS</b>Tamoxifen leads to apoptosis of the cells and reduction in survivin expression, as well as a dramatic reduction in the activated form of p70S6K. Treating HepG2 cells with rapamycin, a specific mTOR inhibitor, significantly reduced the survivin protein level but not affected the survivin transcription, indicating that tamoxifen and rapamycin were synergistic in regards to down-regulation of survivin expression in hepatocellular carcinoma cells.</p><p><b>CONCLUSIONS</b>Our results suggest that tamoxifen down-regulates survivin expression in HepG2 cells and it is mediated by transcriptional and post-transcriptional level via PI3K/Akt/mTOR pathway to induce apoptosis.</p>

Endovascular embolization and microsurgical operation treatment of intracranial anterior circulation aneurysms / 中南大学学报(医学版)

OBJECTIVE@#To explore the surgical treatment and effect of intracranial anterior circulation aneurysms.@*METHODS@#Thirty-eight patients with intracranial anterior circulation aneurysms were enrolled, 9 were treated with endovascular embolization,and 29 with pterion approach micro-euthyphoria operation.@*RESULTS@#One patient was postoperative death. Thirty-four patients were followed up. Among them, 26 were recovery, 1 was botan animation, 2 were meta-palsy, 3 oculomotor palsy, and 2 epilepsy.@*CONCLUSION@#Surgical treatment of intracranial anterior circulation aneurysms is the first choice to help blood tumor cleaning-up and intracranial pressure degrading. Embolotherapy can be applied for patients unfit for operation, but we do not recommend wide use of it due to preoperative cranial nerve injury.

hTERT promoter enhances the radiosensitivity to gene-radiation therapy of human laryngeal carcinoma transplanted in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the therapeutic efficiency of human telomerase reverse transcriptase promoter (hTERTp) mediated horseradish peroxidase (HRP) catalyzed effects of indole-3-acetic (IAA) on laryngeal squamous cell carcinoma with different radiosensitivity in vivo.</p><p><b>METHODS</b>Human laryngeal squamous cell carcinoma Hep-2 and Hep-2R cells were transplanted into nude mice. After growing to about 30 approximately 50 mm3, the tumor-bearing mice were randomly divided into eight groups: Hep-2 line: combined group (A), gene group (B), radiation group (C) and blank group (D); Hep-2R line: combined group (AR), gene group (BR), radiation group (CR) and blank group (DR). The phTERTp-HRP was delivered by intratumoral injection and the IAA by intraperitoneal injection, combined with 2 Gy daily radiation to a total dose of 30 Gy. The tumor volume was recorded. The cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The expression of HRP protein was detected by AP immunohistochemisty.</p><p><b>RESULTS</b>The tumor growth of combined groups was attenuated significantly and the tumor volume of Hep-2R blank group was the largest. The inhibition rate of each group was: A: 54.8%, B: 10.0%, C: 31.9%; AR: 52.7%, BR: 24.8%, CR: 17.0%. In the combined groups, necrosis and apoptosis of tumor cells were observed under the light microscope and the apoptotic index [A (16.6 +/- 1.3)% vs. AR (17.6 +/- 1.3)%] of tumor cells was highest (P < 0.05). The HRP protein expression of BR (33.3 +/- 8.9)% was higher than that of B (21.9 +/- 5.7)%, which was directly up-regulated in the tumors (45.0% vs. 54.8%, P < 0.05) after radiation.</p><p><b>CONCLUSION</b>In the Hep-2- and Hep-2R-transplantation tumors in nude mice, hTERTp can be induced by radiation and enhance the expression of horseradish peroxidase (HRP) gene according to telomerase activity. hTERTp-HRP/IAA system, which has synergistic effects with radiation and inhibits the tumor growth by induction of apoptosis and necrosis, may be a new gene-radiation strategy for the treatment of laryngeal carcinoma.</p>

Insulin/PI3K signalling pathway regulates the expression of survivin in liver cancer HepG2 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To determine the expression level changes of survivin, a inhibitor of apoptosis protein, followed by activation of insulin receptors in human hepatocellular carcinoma HepG2 cell line, and to investigate the signalling pathway involved in the regulation.</p><p><b>METHODS</b>Human hepatocellular carcinoma HepG2 cells were treated with insulin alone or pre-treated with LY294002, a specific inhibitor of PI3K signalling pathway, to determine whether blocking PI3K signaling can attenuate the up-regulation of survivin expression. Real time RT-PCR and Western blot analysis were used to measure survivin mRNA and protein changes before and after treatment, respectively.</p><p><b>RESULTS</b>Without serum supplement, HepG2 cells expressed a small amount of survivin. Insulin induced survivin expression in a dose- and time-dependent fashion. Survivin expression was blocked if cells were pre-treated with LY294002 prior to insulin stimulation.</p><p><b>CONCLUSION</b>Insulin induces survivin expression via PI3K signalling pathway, suggesting that to interfere the key gene in this signalling pathway may block survivin expression, therefore, promoting apoptosis in hepatocellular carcinoma cells.</p>

The influence of hepatitis B e antigen on the expression of toll-like receptor 2 on peripheral monocytes / 中华肝脏病杂志

<p><b>OBJECTIVES</b>In order to investigate the relationship among the toll-like receptor 2 (TLR2), hepatitis B e antigen and HBV DNA, the expression levels of TLR2 on peripheral blood monocytes of chronic hepatitis B (CHB) patients as well as on their monocytes stimulated by ligand of TLR2 (Pam3CSK4) and HBeAg were analyzed.</p><p><b>METHODS</b>Sixty-eight adults with CHB were enrolled, including 37 HBeAg-positive patients, 17 HBeAg-negative and HBV DNA negative patients, and 14 HBeAg-negative and HBV DNA positive patients. Sixteen healthy volunteers were also studied as controls. TLR2 expression levels on their peripheral blood monocytes stimulated with Pam3CSK4 or not stimulated were analyzed by FACS Caliber. The relationship of the expression levels of TLR2, HBeAg and HBV DNA were also analyzed. The level of TLR2 on peripheral blood monocytes of healthy volunteers and HBeAg-negative CHB patients stimulated by HBeAg was examined for six hours.</p><p><b>RESULTS</b>The TLR2 expression levels on CD14+ cells were significantly reduced in HBeAg-positive patients (47.57%+/-21.40 %) compared to both healthy volunteers (76.51%+/-7.46%) and HBeAg-negative patients (HBV DNA positive group 73.2%+/-14.2%, HBV DNA negative group 75.2%+/-11.3%); but there was no difference between those of the HBeAg-negative patients and the healthy volunteers. Expression levels of TLR2 on monocytes stimulated by TLR2 ligand in HBeAg-positive patients were obviously increased, and reached the basic levels of the healthy volunteers and the HBeAg-negative patients. The expression levels of TLR2 on monocytes stimulated by HBeAg of the healthy volunteers and the HBeAg-negative patients were markedly reduced.</p><p><b>CONCLUSIONS</b>In the presence of HBeAg, HBV down-regulates the expressions of TLR2 on CD14+ cells from peripheral blood, and there is no correlation between HBV-DNA and TLR2. Pam3CSK4 can boost the TLR2 expression in HBeAg-positive patients. The proposed interaction between HBV and TLR2 may provide an important clue to explain the reasons of the establishment of persistent HBV infection.</p>