Objective: The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods: The eukaryocyte expressing plasmid pcDNA3.1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3. 1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results: We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion: The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.

Objective: The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods: The eukaryocyte expressing plasmid pcDNA3.1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3. 1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results: We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion: The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.

The 23 cases with chronic myelogenousleukemia (CML) were treated with program ofharringtonine and ara-c (HA) and the results werethat the 13 cases got complete remission and 10cases got partial remission. Comparing with thecontrol group the patients of which were treatedwith busulfan, the authors found that the spendedtime of decreased WBC in blood (WBC decreasedto 10. 0 ? 10~9/L) and reduced spleen (reduced tosmallest status of spleen) and improvement of self-sensation symptoms in the group treated with HAwere better than those in control group. The HAprogram for cases who recured CML again and re-ceived maintenance cure after remission of CMLwas still effective. The time of maintenance cureand interval should be individualized.

A chiral LC-MS/MS method for the simultaneous analysis of desvenlafaxine (DVS) enantiomers in human plasma was developed and applied to a pharmacokinetic study on 12 Chinese healthy volunteers. d6-Desvenlafaxine was used as internal standard (IS). Chromatographic separation was performed on the Astec Chirobiotic V chiral column (150 mm x 4.6 mm, 5 μm). The assay was linear over the concentration range of 0.500-150 ng x mL(-1) for both enantiomers (r2 > 0.99). The method was successfully applied to a stereoselective pharmacokinetic study of 100 mg desvenlafaxine sustained release tablets on 12 Chinese healthy volunteers under fasting conditions. The results showed that the pharmacokinetic parameters were similar to both enantiomers in Chinese healthy volunteers. The AUC(0-t), and C(max) of the two enantiomers were about 1.5 times higher than those of blacks and whites reported in the literature.

OBJECTIVETo observe the impact of electroacupuncture (EA) on the quality of life in the patients of female mixed urinary incontinence (MUI) and evaluate its clinical efficacy.

METHODSEA was adopted in 25 cases of MUI at Zhongliao (BL 33) and Huiyang (BL 35), needle retaining for 30 min each time. The treatment was given once every two days, three treatments a week, totally for 12 weeks of treatment. The incontinence quality of life questionnaire (I-QOL), the incontinence impact questionnaire-7 (IIQ-7), the urogenital distress inventory short form (UDI-SF) and the visual analogue scale (VAS) were adopted in the comparison before and after treatment. The efficacy was evaluated.

RESULTSThe curative rate was 52.0% (13/25) and the total effective rate was 96.0% (24/25) in 25 cases. In comparison before and after treatment, in I-QOL, the score of limiting behaviors was increased up to (30.32 ± 3.39) from (11.52 ± 2.45), the score of psychosocial impacts was up to (36.48 ± 2.90) from (13.52 ± 2.25) and the score of social isolation was up to (21.52 ± 1.61) from (9.84 ± 2.23). The score of IIQ-7 was decreased, down to (2.48 ± 1.36) from (11.96 ± 3.45), the score of UDI-SF was down to (2.92 ± 0.95) from (8.84 ± 2.58) and VAS score was down to (1.40 ± 0.91) from (6.96 ± 1.65). The differences were all significant statistically in comparison (all P < 0.05).

CONCLUSIONEA improves effectively the quality of life in MUI patients and the clinical efficacy is apparent.

Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Middle Aged ; Quality of Life ; Treatment Outcome ; Urinary Incontinence ; physiopathology ; therapy ; Urination

We evaluated clinical application effect of gene chip for detection of rifampin (RFP) and isoniazid (INH) resistance in Mycobacterium tuberculosis (MTB).Rifampin and isoniazid drug-resistance gene loci were detected by gene chip with sputum specimens from smear-positive tuberculosis patients and clinical strains,comparing the results of detection.BACTEC MGIT 960 drug susceptibility test results were used as control to evaluate the detection performance of gene chip.The sequences of the polymerase chain reaction products of the rpoB,katG and inhA genes from 999 strains identified as Mycobacterium tuberculosis were determined to confirm the mutations by DNA sequencing.Results showed that 100 cases were identified as nontuberculous mycobacteria by gene chip in the 1 108 cases of smear-positive samples.Among the rest 1 008 samples,there were only 9 cases of microarray results different from BACTEC MGIT960 culture-positive strains,achieving the coincidences of 99.1％.Compared with BACTEC MGIT 960 drug susceptibility test results,the gene chip method displayed a concordance of 98.1 ％ and 94.5 ％ for RFP and INH respectively in the 999 strains.Compared with the DNA sequencing method,the accuracy of gene chip method was 99.6％ for rifampin resistance and 99.8％ for isoniazid resistance.It's concluded that the gene chip technology can quickly and accurately detect rifampin and isoniazid resistance in MTB and can be used directly for the detection of sputum samples.

Objective To explore the action mechanism of shenxiong glucose injection in treatment of acute spinal cord injury(SCI) through observing its effects on the recovery of motor function and the expression of aquaporin-4(AQP-4) in SCI rats.Methods Totally 90 healthy,aduh,Sprague-Dawley rats were randomly divided into a sham operation group (n=30),aSCI group (n=30) and a drug group (n=30).The SCI rat models in both the SCI group and the drup group were established aecording to the modified Allen's method,while the sham operation group was only given laminectomy.After the operation,the drug group was given intraperitoneal shenxiong glucose injection of 30 rnl/kg a day,while the other two groups were injected in the same way with normal saline.The neural function recovery,the pathological changes after SCI and the expressions of AQP-4 were observed 1,3,7,14 and 21 d after the operation using the Tarlov score,the hematoxylin and eosin staining,as well as immunofluorescence techniques and Western blotting.And the correlation of Tarlov scores with AQP-4 expressions was analyzed.Results No significant changes in Tarlov scores were observed in the sham operation group (P ＞ 0.05),while in the SCI group and the drug group,postoperative Tarlov scores decreased significantly.The hindlimb nerve function recovered to some degree with time in the SCI group and drug group.At 3,7,14 and 21 days after the operation,the Tarlov scores in the drug group were significantly higher than the SCI group (P ＜ 0.05).The drug group showed less severe pathological changes,with more residual neurons still visible of nucleoli than the SCI group 21 days after the operation.Compared with the sham operation group,the expression levels of AQP-4 were significantly higher in the SCI group and drug group at all the time points (P ＜ 0.05).However,the expression levels of AQP-4 in the drug group were significantly lower than the SCI group accordingly (P ＜ 0.05).The Tarlov scores were found to be significantly and negatively related to the AQP-4 protein expression levels 3 days(r =-0.523,P =0.003),7 days(r =-0.437,P=0.016),14 days(r=-0.417,P=0.022) and 21 days(r=-0.377,P=0.040)after the operation.Conclusion Injecting shenxiong with glucose can effectively promote the recovery of motor function after SCI,at least in rats.And its mechanism may be that the development of spinal cord edema is prevented and the secondary spinal cord injury alleviated by restraining the expressions of AQ P-4 in the injured areas.

AIM To investigate the anticonvulsive action of supercritical CO2 ethanol extract from Pinellia Pedatisecta Schott(SEE-CO2PP). METHODS The rat convulsive model was induced by penicillin localized injected in rat cortex. The effects of SEE-CO2PP on the latency of seizure and changes of convulsive behaviors were investigated. The latency of epileptiform discharge, and frequency and amplitude of highest wave in cortex and hippocampus were recorded by using RM6240C multichannel physiological signal collection and analysis recorder. At the same time, the contents of glutamic acid (Glu), aspartic acid (Asp), glycine (Gly) and γ-aminobutyric acid (GABA) in hippocampus were determined with high performance liquid chromatography. RESULTS SEE-CO2PP 15 and 30 g·kg-1, ig, prolonged the latent period of seizure and weakened the extent. SEE-CO2PP also prolonged the latent period of epileptiform discharge, reduced the frequency and decreased amplitude of the highest wave in both cortex and hippocampus. Moreover, SEE-CO2PP increased the content of GABA in hippocampus, but the levels of Gly,Asp and Glu had no obvious changes. CONCLUSION SEE-CO2PP inhibits the epileptiform discharge and convulsive behaviors of convulsive model rats, which suggests SEE-CO2PP has anticonvulsive action.

ObjectiveTo compare the effects of Qishen Fukang capsules and Fluoxetine on cognitive function in first-episode depressions.MethodsBetween December 2011 and January 2012,63 depressed patients with first-episode from Center for Mental Disease Control and Prevention of Third Hospital of PLA were enrolled,and were divided randomly into the Qishen Fukang capsules-treated group ( Chinese medicine group,CMG,31 cases) and fluoxetine treated group (Western medicine group,WMG,32 cases; Jiangsu changzhou pharmaceutical Co.,LTD production) by the method of the random number table.The therapeutic dose was oral 0.2～0.6 g (three times/d) for each CMG patient,and morning oral 20～40 (20 ± 5) mg/d for each WMG patient.Meanwhile,each patient was given the short - term small dosesof benzodiazepine drugs,but no other antidepressants.Each patient and control was assessed with 17-item Hamilton Depression Scale (HAMD).A total of 32 healthy subjects were involved as control group.Each patient was measured with evoked event-related potential P300 change before and after 6 weeks treatment.And compared the effects of Qishen Fukang capsules and Fluoxetine on cognitive function in first-episode depressions before and after treatment.Results(1) Compared with prior-treatment [ CMG ( 29.1 ± 5.1 )score vs WMG(29.0 ± 4.5)score],the HAMD scores of post-treatment [ CMG( 10.1 ± 3.2) score vs WMG (12.3 ± 3.4) score] were decreased significantly ( P＜0.05).The HAMD scores in CMG were significantly lower than the WMG( P＜0.05).The HAMD reductive rate in CMG was significantly higher than the WMG [ (65.6 ± 2.1 ) ％ vs (57.9 ± 3.2 ) ％,P ＜ 0.05 ].(2)compared with prior-treatment,the latency periods of post-treatment were shortened[ P2 (152.8 ± 54.1)ms vs (208.9 ± 57.6)ms,(174.5 ±63.2)ms vs (207.3 ± 55.8) ms;N2(208.7 ± 57.9)ms vs (273.4 ± 62.0) ms,(239.2 ± 59.2) ms vs (275.6 ± 60.8)ms; P3(319.1 ±60.2)ms vs (396.3 ± 66.3)ms,(315.6 ± 61.1)ms vs (394.7 ±55.6)ms ],while the amplitudes were prolonged [ P2 (7.8 ± 1.7 ) μV vs ( 3.3 ± 1.2 ) μV,( 7.0 ± 1.4 ) μV vs　(3.4±1.4)μV; N2 (3.6±1.4)μV vs (1,0±0.7)μV,(2.4±1.3)μV vs (1.2 ± 1.0)μV; P3 (9.6±2.2)μV vs (4.5 ± 1.0)μV,(7.5 ±2.2)μV vs (4.6 ± 1.2)μV] in the CMG and WMG (all　P ＜0.05).Compared with the CMG,the latency periods of F2,and N2 were signiflcantly longer (all P ＜ 0.05 ),and the latency period of p3 was no difference ( P＞0.05 ),and the amplitudes of N2,and P3 were lower ( all P ＜ 0.05 ),and the amplitude of P2 was no difference ( P＞0.05 ) in the post-treatment of WMG.(3)Compared with the controls,the latency periods of P2,N2,and P3 were shortened,while the amplitudes were prolonged in the post-treatment of CMG and WMG ( all P ＜ 0.05 ).The latency periods and amplitudes of P2,N2,and P3 did not show any difference in the CMG after treatment ( all P ＞ 0.05).The latency period of P2,and N2 were still significantly longer (all＜0.05) and the latency period of P3 was no difference ( P＞0.05) while the amplitude of P2,N2,and P3 were still significantly lower (all P ＜ 0.05) in the post-treatment of WMG.ConclusionsQishen Fukang capsules and Fluoxetine can improve significantly cognitive function in first-episode depressions.Qishen Fukang capsules is superior to fluoxetine on improving the early preparation efficiency of information processing.