Value of Serum Light Chain in Diagnosis and Evaluation of Efficacy for Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical application value of serum light chain (sLC) in the diagnosis and therapeutic efficacy evaluation for multiple myeloma.@*METHODS@#46 patients with newly diagnosed multiple myeloma were selected as MM group and 50 healthy persons as control group. Rate scattering immunoturbidimetry was used to detect serum light chain and immunoglobulin (Ig) in two groups, serum protein electrophoresis was used to detect M protein by agarose gel. Then, the sensitivity and specificity of the two methods in MM diagnosis were analyzed and compared, and the significance of sLC detection in MM diagnosis were discussed. In addition, 15 MM patients after received conventional therapy were tracked, sLC levels in five different therapentic times were recorded, and the effect of sLC in efficacy evaluation of MM was analyzed.@*RESULTS@#There were 11 cases of IgA type, 15 cases of IgG type, 8 cases of light chain κ type, 8 cases of light chain λ type, 2 cases of IgD type, and 2 cases of non-secretion type. The sLC-κ, sLC-λ and their ratio (including light chain type and double clone type), IgA and IgG (except IgD type), as well as albumin, beta-globulin and gamma-globulin levels showed statistically significant differences (P＜0.05) compared with the control group. The sensitivity of serum protein electrophoresis, Ig quantification, sLC and its ratio in the diagnosis of multiple myeloma were 57%, 76% and 65%, and their specificity were 83%, 61% and 90%, respectively. After the second or third chemotherapy, the sLC-κ/λ ratio gradually approached the normal range as the disease reliefes, and the sLC-κ/λ ratio continued to be on or off the line at outliers or further away from the reference value as the disease progresses in MM patients with κ type or λ type.@*CONCLUSION@#sLC detection shows positive significance in early diagnosis of multiple myeloma, SLC monitoring can be used for the efficacy evaluation for treatment of MM patients.

Twist promotes cell migration and invasion in human breast cancer cell line Hs578T / 基础医学与临床

Objective To study the effects of Twist gene silencing on migration and invasion of human triple-nega-tive breast cancer cell Hs578T and its potential mechanisms. Methods The lentiviral vectors with Twist gene-targe-ted specific shRNA and the negative control were constructed,then transfected into the human triple-negative breast cancer cell line Hs578T to establish the stable cell lines of Twist gene silencing. The blank group(blank),negative control group(shNC) and Twist gene silencing group(shTwist) were set up.After screening by puromycin,the fluo-rescence expression and the infection efficiency of each group cells were observed by inverted fluorescence micro-scope. The expression of Twist mRNA and protein level was detected by quantitative real-time PCR and Western blot.Transwell migration and invasion experiments were performed to measure cell migration and invasion abilities, re-spectively. Western blot was used to detect the levels of p-AKT, AKT, p-ERK1/2 and ERK1/2 proteins. Results Twist gene was successfully knockdown in human triple-negative breast cancer cell line Hs578T. Compared with the blank group and shNC group cells,the cell migration and invasion ability of the shTwist group cells were decreased significantly(P<0.05,P<0.05),and the expression of Twist downstream p-AKT and p-ERK1/2 proteins were al-so reduced notably in the shTwist group cells(P<0.05,P<0.05). Conclusions Twist promotes cell migration and invasion via AKT/ERK signaling axis in human triple-negative breast cancer cell line Hs578T.

Construction of a lentiviral vector carrying short?hairpin RNA targeting PAX6 and its effect on proliferation of glioma U251 cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral vector for delivering short hairpin RNA (shRNA) targeting PAX6 and investigate its effect on the proliferation of glioma U251 cells in vitro.</p><p><b>METHODS</b>Two small interfering RNA sequences targeting PAX6 gene were designed based on the reported sequence of PAX6 and annealed to form a double?stranded chain, which was inserted into a lentiviral vector to construct the recombinant lentiviral vector shRNA?PAX6. The recombinant vector was infected into U251 cells, and the expression of PAX6 mRNA and protein in the cells was detected by real?time PCR and Western blotting, respectively. The changes in the proliferation of U251 cells after the infection was assessed using MTT assay.</p><p><b>RESULTS</b>Double enzyme digestion of the lentiviral vector pLKD?CMV?G&NR?U6?shRNA yielded an 8208?bp fragment, and colony PCR and sequencing analysis confirmed successful construction of the lentiviral vector shRNA?PAX6. Infection of the cells with shRNA?PAX6 caused a significant reduction of the expressions of PAX6 mRNA and protein (P<0.05) and resulted in obviously increased proliferation of U251 cells (P<0.05).</p><p><b>CONCLUSION</b>We successfully constructed the recombinant vector shRNA?PAX6 for silencing PAX6 gene. PAX6 gene silencing results in increased proliferation of U251 cells in vitro.</p>

A cross-sectional study on the effect of virological response after HAART on subsets of T lymphocytes and expression of CD127 in pediatric AIDS patients with different viral loads / 中华儿科杂志

<p><b>OBJECTIVES</b>To study the effect of HAART on subsets of T lymphocytes and expression of CD127 on memory and naїve CD4(+) and CD8(+)T cells in pediatric AIDS patients with different viral loads receiving HAART.</p><p><b>METHOD</b>A cross- sectional study on 194 pediatric AIDS patients receiving HAART was carried out and 52 age matched healthy children were recruited as controls. The percentage of CD4(+), CD8(+), CD8(+)CD45RA(+)CD127(+/-), CD8(+)CD45RO(+)CD127(+/-), CD4(+)CD45RA(+)CD127(+/-) and CD4(+)CD45RO(+)CD127(+/-)T cells was tested using flow cytometry, and HIV-RNA in plasma was detected by quantitative RT-PCR.</p><p><b>RESULT</b>The percentage of memory (CD45RO(+)) CD4(+)T cells decreased to (45.73 ± 8.85)%, and that of naїve (CD45RA(+)) CD4(+) and memory CD8(+)T increased to (60.44 ± 5.01)% and (54.69 ± 7.71) % respectively in the pediatric AIDS patients vs. controls (P < 0.05). The percentage of naїve (CD45RA(+)) CD4(+)T cells of patients with viral load (VL) < 400 copies/ml was (65.57 ± 5.33) %, which was significantly higher than that of patients with VL ≥ 400 copies/ml (P < 0.05).Of patients with VL < 400 copies/ml, the percentage of CD4(+)CD127(+)T cells, especially the subset of memory CD4(+)CD127(+)T cells was (82.35 ± 2.31)%, which was higher than that of patients with VL ≥ 400 copies/ml, but lower than that of controls (P < 0.05). The percentage of memory and naїve CD8(+)CD127(+)T cells was lower than that of controls (P < 0.05).</p><p><b>CONCLUSION</b>The recovery of CD4(+)T cell subsets in pediatric AIDS patients is associated with viral load. Effective HAART can increase the percentage of naїve CD4(+)T cells and the life of memory CD4(+)T cells.</p>

Significance of sentinel lymph node detection for cN0 laryngeal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The purpose of this study was to investigate the clinical value of radiolabeled tracer method, methylene blue method and combination of these two methods in detection of sentinel lymph node (SLN), and to evaluate the accuracy of SLN in predicting the cervical lymph nodes status in laryngeal carcinoma patients with clinically negative neck lymph nodes (cN0 ).</p><p><b>METHODS</b>Forty-one patients with cN0 laryngeal neoplasms underwent SLN detection using both of radiolabeled tracer and methylene blue. SLN imaging was performed with laryngoscope-guided injection of radioactive isotope 99Tc(m)-sulfur colloid (SC) into the laryngeal carcinoma before surgery, then all these patients underwent intraoperative lymphatic mapping with a handheld gamma-detecting probe. After mapping of SLN, methylene blue was subsequently injected at the same spots around the tumor in order to identify SLN during surgery. The results of SLN detection by isotope tracer, dye and combination of both methods were compared.</p><p><b>RESULTS</b>The SLN detection rates by radiolabeled tracer, methylene blue and combined method were 87.8%, 70.7% and 92.7%, respectively (P < 0.01). The number of detected SLN was significantly different between radiolabeled tracer method and combined method (P < 0.05), and also between blue dye method and combined method (P < 0.01). However, no statistically significant difference was found between methylene blue method and radiolabeled tracer method (P > 0.05). Nine patients were found to have lymph node metastasis by final pathological examination. The sensitivity, accuracy and negative predictive values of SLN detection by the combined method using radiolabeled tracer and methylene blue were 88.9%, 97.4% and 96.7%, respectively.</p><p><b>CONCLUSION</b>The combined method using radiolabeled tracer and methylene blue can improve the accuracy of sentinel lymph node detection. Furthermore, sentinel lymph node detection can accurately predict the cervical lymph node status in cN0 laryngeal carcinoma.</p>

Establishment and evaluation of a bone cancer pain model / 中南大学学报(医学版)

OBJECTIVE@#To explore the feasibility of a bone cancer pain model by injecting the Lewis lung carcinoma cells into the femur bone marrow cavity of C57BL/6 mice.@*METHODS@#Sixty clear grade male C57BL/6 mice (body weight 18 approximately 20 g) were randomly divided into 4 groups(15 in each group). Cancer cell inoculated group: 2*10(6) Lewis lung carcinoma cells in 10 microL PBS were injected into the left femur bone marrow cavity, and the other 3 control groups were injected the heat inactivated Lewis cells, PBS, or a false operation respectively. Spontaneous lifting time and mechanical allodynia threshold of the mice hind paw were measured in the alternative days throughout the experiment. The structural damage of the femur was monitored by radiogram on the 7th,15th, and 23rd day respectively,and the pathohistological changes of the femur bones were observed by HE staining on the same days.@*RESULTS@#Those mice that received intra-femur innoculation of Lewis lung carcinoma cells gradually developed the spontaneous pain, which was began on the 11th day after the innoculation, and followed by mechanical allodynia. The course of flinch lasted in the later experimental session. The 50% Von Frey threshold was significantly decreased on the 13th day after the innoculation, and the mechanical allodynia lasted the whole experimental period. On the 23rd day after the innoculation, X-ray film showed that the medullary cavity of ipsilateral distal femur was filled with tumor cells, and the cortical bone became thick; furthermore, the tumor cells invaded the peripheral muscles.@*CONCLUSION@#Injecting the Lewis lung carcinoma cells into the femoral medullary cavity of C57BL/6 mice can successfully establish a murine bone cancer pain model, and the murine model shows much resemblance compared with the human bone cancer pain.

Effects of hypoxic preconditioning on hypoxia tolerance of astrocytes in vitro / 中国应用生理学杂志

<p><b>AIM</b>To explore the mechanisms of hypoxic preconditioning on protecting cultured astrocytes from hypoxia injury.</p><p><b>METHODS</b>Cultured astrocytes were divided randomly into several groups: control(C), hypoxia(H) and hypoxic preconditioning (HP). Cells MTT metabolic activity, qualitation of apoptosis and modality to explore the protection effects of hypoxic preconditioning. Immunocytochemistry of Bcl-2 and Bax to explore the mechanisms of hypoxic preconditioning on protecting astrocytes from hypoxia.</p><p><b>RESULTS</b>Compared with H group there was marked increase of MTT metabolic activity in HP48 and HP72 groups. Immunocytochemistry of Bcl-2 and Bax showed that compared with H group, expression of Bcl-2 was increased in HP group, while expression of Bax was decreased in HP group.</p><p><b>CONCLUSION</b>Hypoxic preconditioning can protect astrocytes from hypoxia. One possible mechanism maybe concerned with inhibition of Bax and maintain of Bcl-2 to depress apoptosis procedure.</p>

Protection of hepatocyte growth factor on neurons subjected to oxygen-glucose deprivation/reperfusion / 生理学报

The present study was conducted to investigate the effect of hepatocyte growth factor (HGF) on cortical neurons exposed to oxygen-glucose deprivation/reperfusion (OGD/R). Primary cultured cerebral cortical neurons were prepared from Sprague-Dawley rats. The cells were used for experiments after culture for 12 d in vitro. To initiate OGD/R, the culture medium was replaced by glucose-free medium, and cells were transferred to a humidified incubation chamber flushed by a gas mixture of 95% N(2) and 5% CO(2) at 37 °C for 2 h. Following this treatment, neurons were fed with glucose-supplemented (25 mmol/L) medium, and returned to the incubator under normoxic condition for 0-24 h. The cell viability was assessed by MTT assay, and cell injury was evaluated by lactate dehydrogenase (LDH) leakage rate. The percentage of apoptotic cells was analyzed by flow cytometry and Hoechst 33258 staining. The expressions of c-Met mRNA and protein were detected by RT-PCR and Western blot analysis, respectively. Oxygen-glucose deprivation for 2 h decreased the cell viability and increased LDH leakage rate in cultured cerebral cortical neurons. The cell viability declined and LDH leakage rate increased with the reperfusion time going on (0-24 h). To explore the influence of HGF on neurons under oxygen-glucose deprivation for 2 h/reperfusion for 24 h (OGD(2)/R(24)) condition, the cultures were pretreated with HGF at different concentrations (5-120 ng/mL) 2 h prior to OGD(2)/R(24). The results showed that OGD(2)/R(24) treatment significantly decreased the cell viability, increased LDH leakage rate and the percentage of apopototic cells. Pretreatment with HGF at 5 ng/mL and 10 ng/mL did not affect the decrease in cell viability resulting from OGD(2)/R(24). In the presence of 20 ng/mL HGF, the increase in cell viability in cortical neurons exposed to OGD(2)/R(24) began to appear, and 80 ng/mL of HGF exhibited the maximal effect. HGF at 5, 10 and 20 ng/mL did not affect the increase in LDH leakage rate in cortical neurons exposed to OGD(2)/R(24). In the presence of 40 ng/mL HGF, the decrease in LDH leakage rate in cortical neurons subjected to OGD(2)/R(24) began to appear, and 80 ng/mL of HGF displayed the maximal effect. In addition, HGF at 80 ng/mL significantly attenuated cell apoptosis resulting from OGD(2)/R(24). As detected by semi-quantitative RT-PCR and Western blot analysis, c-Met mRNA and protein were expressed in cerebral cortical neurons cultured for 12 d in vitro. c-Met mRNA and protein expressions in cortical neurons exposed to OGD(2)/R(24) were significantly upregulated and were not affected by pretreatment of HGF at 80 ng/mL. Treatment with c-Met inhibitor SU11274 (5 μmol/L) completely eliminated HGF-mediated protection of cortical neurons subjected to OGD(2)/R(24). The results suggest that HGF directly protects cortical neurons against OGD/R-induced cell injury in a dose-dependent manner, and HGF has a potent anti-apoptotic action on neurons exposed to OGD/R.

Effect of thrombin on cultured rat cerebral astrocyte injured by hypoxia/reoxygenation and its relationship with iNOS / 中南大学学报(医学版)

OBJECTIVE@#To observe the effect of thrombin on the cytotoxicity of astrocytes injured by hypoxia/reoxygenation(H/R) and to explore its relationship with inducible nitric oxide synthase (iNOS).@*METHODS@#Primary astrocytes were cultured in DMEM with 10% approximately 15% calf serum and divided into 6 groups: a control group, a Tm control group, an H/R group, a Tm+H/R group, a hirudin (HR) control group, and a Tm+HR+ H/R group. The cell damage and viability were detected by the 3-(4, 5-di-methyl-thazol-2-yl)-2, 5 diphenyl-tetrazol-iumbromide (MTT) conversion method. The NO level in the cultured cell supernatant was assayed by Griess reagent. The flow cytometry was performed to evaluate the apoptosis rate of astrocytes. The iNOS mRNA was examined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Immunocytochemistry was used to observe the expression of iNOS protein.@*RESULTS@#The cell viability injured by H/R was lower than that of the control group, the NO production and apoptosis rate in the cell of H/R group were higher than those of the control group. Incubation of H/R cell with 10kU/L Tm enhanced the cytotoxicity of H/R stimulation compared with the cells injured by H/R. Hirudin can reverse the effect of thrombin. RT-PCR and immunocytochemistry analysis demonstrated that the levels of iNOS mRNA and iNOS protein increased in the cells treated by H/R. Tm enhanced the expression of iNOS mRNA and iNOS protein in the cells treated by H/R. Hirudin blocked the effect of Tm.@*CONCLUSION@#Increasing the level of iNOS and enhancing the production of NO may be the mechanism of thrombin cytotoxicity in astrocytes injured by H/R.

Protective effect of thrombin precondition on astrocytes insulted by oxygen-glucose deprivation / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of thrombin precondition (TPC) on the rat cerebral astrocytes(As) cultured in oxygen-glucose deprivation (OGD).@*METHODS@#Astrocytes were pretreated with thrombin (TB) at various concentrations (0.005 approximately 5.000 kU/L), and then insulted by OGD. The cell damage and viability were evaluated by the lactate dehydrogenase (LDH) effusion rate and the 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) conversion method. Detection of apoptotic cells was determined by the flow cytometry technique. The glutamate uptake of astrocytes was studied with [3H]-glutamate incorporation.@*RESULTS@#OGD increased the LDH, decreased the cell viability, increased the number of apoptotic astrocytes, and decreased the glutamate uptake (P<0.01). While preconditioned with thrombin at the same condition, the LDH decreased, the cell viability increased, the percentage of apoptotic cells decreased, and the glutamate uptake increased (P<0.05). The maximum protective effect of thrombin was observed at 0.1 kU/L.@*CONCLUSION@#Low concentration of thrombin precondition (TPC) can protect the astrocytes from oxygen-glucose deprived injury, and attenuate its apoptosis in a dose-dependent manner.

MICA*008/A5.1 allele and HCMV infection in kidney transplanted donees of Hunan Han nationality / 中南大学学报(医学版)

OBJECTIVE@#To investigate the relationship between MICA*008/A5.1 allele and human cytomegalovirus (HCMV) infection in kidney transplanted donees of Hunan Han nationality.@*METHODS@#The MICA*008/A5.1 allele based on 91 kidney transplanted donees and 81 unrelated normal individuals of Han nationality in Hunan Province were analyzed by PCR/SSP assay. At the same time, anti-HCMV antibody IgM was detected in the serum by ELISA method.@*RESULTS@#The positive rate of MICA*008/A5.1 allele was significantly higher in the control group (56.79%) than that in the kidney transplanted donee group (34.07%) (P <0.05). The infection rate of HCMV in those individuals whose genotype was MICA*008/A5.1 (-) was significantly higher than that in the MICA*008/A5.1(+).@*CONCLUSION@#The individual whose genotype is MICA*008/A5.1 (+) is not liable to HCMV infection, but the individual whose genotype is MICA*008/A5.1 (-) is liable to HCMV infection.

Effect of hepatocyte growth factor on oxygen-glucose deprived injury of astrocytes / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of hepatocyte growth factor (HGF) on oxygen-glucose deprived injury and apoptosis of astrocytes.@*METHODS@#The injury of primary cultured rat cerebral cortical astrocytes was induced by oxygen-glucose deprivation. Astrocytes were treated with HGF at various final concentrations of 20 - 100 ng/mL. The cell damage and viability were evaluated by the lactate dehydrogenase (LDH) released rate and the 3- (4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) conversion method. Detection of apoptotic cells was determined by the flow cytometry, and the ultrastructure was observed by the transmission electron microscope.@*RESULTS@#Oxygen-glucose deprivation increased the LDH release rate, decreased the cell viability and increased the number of apoptotic astrocytes. While exposed to HGF at the same condition, the LDH release rate decreased, the cell viability increased, and the percentage of apoptotic cells decreased (P <0.05). The maximum protective effect of HGF was observed at 60 ng/mL.@*CONCLUSION@#HGF can protect cultured astrocytes from oxygen-glucose deprived injury, and attenuate the apoptosis of astrocytes in a dose-dependent manner.