Chimeric antigen receptor T-cell (CAR-T) technology is based on genetic modification technology to express T-cell expression tumor specific chimeric antigen receptor bind tumor antigen in an antigen-dependent anti-MCH way. Single chain antibody fragment (scFv) of tumor-associated antigen (TAA) combines with up-stream activating sequence of T-cell in vitro. The forming recombinant plasmid transfects the purified and large scale proliferating T-cell in vitro by transfection technique. This process starts and activates specific killing reaction of tumor. The clinical application of cell therapy shows high efficiency and good anti-tumor effect in treatment of malignant neoplasm, such as leukemia, lymphoma, melanoma, which made CAR-T become the mainstream method of cell therapy.

Objective To design and identify small interfering RNA (siRNA) targeting tumor necrosis factor‐α (TNF‐α) ex‐pression ,siRNAs were electroporated into synovial cells of Human to screen those which can effectively suppress TNF‐α expres‐sion .Methods Three TNF‐α specific double stranded siRNA were designed targeting different regions of TNF‐α mRNA(compared with negative control group) .RT‐PCR and Elisa were applied to detect TNF‐α ,mRNA expression and the secretion of TNF‐α in cell supernatant ,respectively .Results Two of the three customized TNF‐α siRNA could inhibit the expression of TNF‐α mRNA in synovial cells of Human (P< 0 .01) .At the same time ,the secretion of TNF‐α decreased in cell supernatant ,the difference was sig‐nificant statistically compared with the control group(P< 0 .01) .Conclusion TNF‐α siRNA can be successfully designed and syn‐thesized ,which can specifically and effectively suppress TNF‐α mRNA expression .

Objective To compare the curative effects of multi-level puncture versus single-level puncture percutaneous vertebroplasty(PVP) for treatment of multiple-level osteoporotic vertebral body compression fractures(OVCF) in senile patients.Methods From June 2008 through November 2014,89 senile patients with fresh multiple-level OVCF underwent PVP guided by C-arm fluoroscopy in prone position.Of them,51 received PVP in which the vertebrae of multiple levels were simultaneously punctured for bone cement injection while the other 38 received PVP in which the vertebrae of multiple levels were punctured one by one for bone cement injection.The 2 groups were compatible with no significant differences in preoperative demographic data (P ＞ 0.05).The 2 groups were compared in terms of visual analgesic scale (VAS),operation time for a single vertebra,fluoroscopy times for a single vertebra,bone cement injection amount for a single vertebra,and extraosseous cement leakage.Results PVP procedures were successful in both groups without serious complications.The VAS scores in both groups at 2 days post-operation were significantly lower than those at pre-operation(P ＜ 0.05).The operation time and fluoroscopy times for a single vertebra in the multi-level puncture PVP group were significantly less than those in the single-level puncture PVP group (P ＜ 0.05).There were no significant differences between the 2 groups in bone cement injection amount for a single vertebra or extraosseous cement leakage (P ＞ 0.05).Conclusions The curative effects of multi-level puncture and single-level PVP are satisfactory for senile OVCF,but multi-level puncture PVP may lead to less operation time and less X-ray exposure.

Objective To determine the dangerous depths of acupuncture at Yamen(GV15) and Fengfu(GV16) in different age groups and different body types based on MRI measurements and to analyze their distribution chracteristic .Methods 98 patients of different age groups in the affiliated hospital of Hainan Medical College from June to December 2012 were selected and the each age group was divided into 3 types :asthenic type ,moderate type and endomorphy type according to the Rohrer index .All images took the median sagittal position .The dangerous depth was measured according to the clinical operation habit and actual feasibility .Re-sults The difference in the acupuncture depth of GV15 and GV16 among different asthenic type patients in different age groups had no statistical significance ;in the moderate type patients ,the difference in the acupuncture depth of GV15 and GV16 between the age group of 18- <45 years and 45-60 years had the statistical significance(P=0 .029 ,P=0 .018);in the endomorphy type pa-tients ,the difference in the acupuncture depth of GV15 and GV16 between the age group of 45-60 years and the other age groups had the statistical significance(P<0 .05) .Conclusion The dangerous depth of GV15 and GV16 had no correlation with age ,but is positively correlated with the Rohrer index ,i .e .,the dangerous depth of GV15 and GV16 is increased with the increase of Rohrer index .

Objective To observe the results and side effects of radiotherapy(RT) combined with hyperthermia(HT) for non-small cell lung cancer(NSCLC)with lesion(T) ≥5?cm in diameter.Methods Patients conforming to T≥5?cm or T volume ≥125?cm~3 in the lung and other required eligibilities were given RT and HT.The temperature of HT in the center of Ts was automatically programmed between 43.0℃ and 43.5℃ by a computer.RT dose of D_T60-66?Gy was delivered in 30-33 fractions with in 1 hour after HT.Results From October 2001 to April 2004,30 patients were registered for this study.Among all patients,23 males and 7 female with median age of 61 years(32-81);22 squamous cancer and 8 adenocarcinoma type;6 stage Ⅰb,1 Ⅱb,10 Ⅲa,8 Ⅲb and 5 Ⅳ lesions.The mean volumes of T was 275.45?cm~3(125-900?cm~3).The mean number of HT given was 7.2 times(2-14).The mean irradiation dose was 59.10?Gy(23.96-9.67?Gy).Five out of 30 patients were dropped from this trial due to reasons un-related to acute side effects of RT or HT.None complained of breathlessness or aggravation of respiratory symptom during heating.No change had been observed in blood pressure,pulse or frequency of breath before or after HT,nor was there any hardening of subcutaneous fatty tissue as well as lipid nodules or severer than grade Ⅲ acute irradiation side effects.Complete response was observed in 23%,partial response in 50%,no change or progression in 27%.Loco-regional control rate was 83% with a median of median locoregional duration of 19.2 months.Eleven patients had died,including 2 of hemoptysis.The median survival duration was 19.7 months and 1-year survival rate was 75%.Conclusions Radiotherapy combined with hyperthermia modal can be used for NSCLC with primary lesion≥5?cm as it carries better efficiency and loco-regional control without server side effects.

Objective Conventional examination for vision function adopts subjective psychophysics methods.Pattern visual evoked potential (P-VEP) binocular summation response is a new objective way for the test of vision function.But its clinical value in evaluating vision function of amblyopia children is still in controversy.This study was to explore the binocular vision and the feature of P-VEP binocular summation in children with amblyopia and evaluate the significance of P-VEP binocular summation in binocular vision.MethodsThis is a case-controlled study.P-VEP binocular summation response and single ocular P-VEP response was respectively recorded in 151 hyperopia amblyopia children and 80 age- and gender-matched normal children.P-VEP response from children with hyperopia amblyopia was recorded under the corrected vision.The hyperopia amblyopia was diagnosed based on the standard of National Children Amblyopia and Strabismus Prevention and Treatment Working Group.The feature of P-VEP binocular summation was further analyzed and compared with monocular VEP response.Informed consent was obtained from each subject or custodian prior to the trail.ResultsNo significant difference was found in latency between P-VEP binocular summation response and monocular P-VEP response whether amblyopia children or normal children(P＞0.05).Binocular response/monocular response value in amblyopia children was lower than that in normal children(P＜0.05).Binocular response/monocular response value in mild or moderate amblyopia children was higher than that in severe amblyopia children(P＜0.05).Binocular response/monocular response value was significantly reduced in the children with binocular vision than in the children without binocular vision (P＜0.05).ConclusionP-VEP binocular vision,acting as an evaluation factor,could impartially reflect the function of binocular vision of amblyopia children.

The shape and structure of granules are controlled by the granulation process, which is one of the main factors to determine the nature of the solid dosage forms. In this article, three kinds of granules of a traditional Chinese medicine for improving appetite and promoting digestion, namely, Jianwei Granules, were prepared using granulation technologies as pendular granulation, high speed stirring granulation, and fluidized bed granulation and the powder properties of them were investigated. Meanwhile, synchrotron radiation X-ray computed micro tomography (SR-µCT) was applied to quantitatively determine the irregular internal structures of the granules. The three-dimensional (3D) structure models were obtained by 3D reconstruction, which were more accurately to characterize the three-dimensional structures of the particles through the quantitative data. The models were also used to quantitatively compare the structural differences of granules prepared by different granulation processes with the same formula, so as to characterize how the production process plays a role in the pharmaceutical behaviors of the granules. To focus on the irregularity of the particle structure, the box counting method was used to calculate the fractal dimensions of the granules. The results showed that the fractal dimension is more sensitive to reflect the minor differences in the structure features than the conventional parameters, and capable to specifically distinct granules in structure. It is proved that the fractal dimension could quantitatively characterize the structural information of irregular granules. It is the first time suggested by our research that the fractal dimension difference (Df,c) between two fractal dimension parameters, namely, the volume matrix fractal dimension and the surface matrix fractal dimension, is a new index to characterize granules with irregular structures and evaluate the effects of production processes on the structures of granules as a new indicator for the granulating process control and optimization.

BACKGROUND:It is unclear about dexamethasone effect on the regulation of microRNA-155 expression in macrophages.
OBJCTIVE:To explore whether dexamethasone can regulate the expression of microRNA-155 in macrophages.
METHODS:(1) Lipopolysaccharide stimulation of mouse macrophages: mouse macrophage cel lines, Raw264.7 cels, were culturedin vitro and stimulated by lipopolysaccharide. Cultured cels were colected at 0, 0.5, 2, 6 hours after culture to detect the dynamical expression of microRNA-155. (2) Dexamethasone intervention for macrophages: Macrophages were divided into four groups: control group treated with phosphate buffer; lipopolysaccharide group stimulated by lipopolysaccharide; combined group given intervention with dexamethasone and lipopolysaccharide; dexamethasone group cultured with dexamethasone. At 6 hours after culture, cel supernatant was colected to detect the expression of tumor necrosis factor α and interleukin-6 using ELISA method. Real-time fluorescence quantitative PCR was used to detect the expression of microRNA-155 in the Raw264.7 macrophages.
RESULTS AND CONCLUSION:Lipopolysaccharide significantly increased the expression of tumor necrosis factor α, interleukin-6 and microRNA-155 after 6 hours of culture (P < 0.05). Combined use of dexamethasone and lipopolysaccharide slightly increased the expression of tumor necrosis factor α, interleukin-6 and microRNA-155 (P< 0.05). Dexamethasone alone had no influence on the expression of tumor necrosis factor α and interleukin-6, but significantly decreased the expression of microRNA-155 (P < 0.05). These findings indicate that dexamethasone can inhibit the expression of microRNA-155 in the macrophages induced by lipopolysaccharide.

Objective To investigate the growth inhibition and DNA damage of energized fusion protein anti-CD20(Fab)-LDM on B JAB cells in vitro.Methods The binding activity of fusion protein anti-CD20 (Fab)-LDP to B JAB cells was studied by flow cytometry and confocal laser scanning microscopy.MTT assay was used to study the energized fusion protein anti-CD20(Fab)-LDM on cell growth of B JAB cells.Comet assay was employed to detect DNA damage in B JAB cells.The cell growth cycle of BJAB was analyzed by FACS.Results The recombinant fusion protein anti-CD20 (Fab)-LDP possessed an significant target affinity towarded BJAB cells.The energized fusion protein anti-CD20(Fab)-LDM showed obvious inhibition on proliferation,as well as induced potent DNA damage in B JAB cells in vitro compared with lidamycin.B JAB cells treated with energized fusion protein anti-CD20 (Fab)-LDM showed S phase cell cycle.Conclusion The energized fusion protein anti-CD20 (Fab)-LDM could target binding to BJAB cells and significantly inhibit the proliferation of B JAB cells by inducing DNA damage and S phase arrest.

OBJECTIVE:To study the effects of simvastatin on oxidative stress and cell apoptosis in aged mice with myocardi-al ischemia-reperfusion (IR). METHODS:Aged mice were randomly divided into sham operation group (phosphate buffer solu-tion),model group(phosphate buffer solution)and simvastatin low-dose,medium-dose and high-dose groups(2.5,5 and 20 mg/kg) with 14 mice in each group. Those groups were given relevant medicine intraperitoneally before modeling for 7 d,once a day. IR model was induced in those groups except for sham operation group. The area ratio of myocardial infarction,myocardial cell apop-tosis rate,activity of myocardial tissue apoptosis gene Caspase-3,the protein expression of Bax and Bcl-2,Akt phosphorylation, serum concent of MDA and activity of SOD were all detected. RESULTS:Compared with sham operation group,the area ratio of myocardial infarction,myocardial cell apoptosis rate,Caspase-3 activity,the protein expression of Bax and MDA content were all increased in model group,while the protein expression of Bcl-2,Akt phosphorylation and SOD activity were decreased(P<0.01). Compared with model group,the area ratio of myocardial infarction,myocardial apoptosis rate,Caspase-3 activity,the protein ex-pression of Bax and MDA content were all decreased in simvastatin high-dose group,while the protein expression of Bcl-2,Akt phosphorylation and SOD activity were increased (P<0.01). There was no statistical significance in above indexes in simvastatin low-dose and medium-dose groups (P>0.05). CONCLUSIONS:Simvastatin can relieve myocardial IR injury in aged mice,and the mechanism of which may be associated with inhibiting myocardial cell apoptosis and the generation of oxidative stress.