Objective To construct a luciferase reporter gene vector of perforin promoter and methylate it in vitro. Methods The promoter of the human peffofin was amplified by PCR, cloned into pMD18-T vector and subcloned into pGL3-Basie vector, and then con-finned by restriction mapping and DNA sequencing. The regions of interest were excised with the appropriate restriction endonucleases, then it were methylated with methylase Sss I(M. SssI)and S-adenosymethioine(SAM), and methylation was confirmed by digestion with appropri-ate methylation sensitive enzyme AciI and agrose gel electrophoresis, and then the fragment was relegated back into the promoter-reporter constructor pGL3-Basic. Results The result of DNA sequencing showed that the sequence of cloned promoter was right. The result of diges-tion methylation with appropriate methylation sensitive enzyme showed that perforin promoter was completely methylated. Conclusion The promoter of perforin was successfully cloned and completely methylated in vitro, which provided an important basis for the study of transfec-tion.

Objective: To observe the efficacy of herbal cake-partitioned moxibustion for lumbar disc herniation (LDH) due to kidney deficiency and blood stasis and observe the influence of this method on lumbar functions and inflammatory factors in patients with this condition. Methods: A total of 120 LDH patients who met the inclusion criteria were randomly divided into three groups, including a herbal cake-partitioned moxibustion group, a flour cake-partitioned moxibustion group, and a Western medication group, with 40 patients in each group. The patients in the Western medication group were treated with diflunisal tablets, 0.5 g per dose, 2 doses a day. Those in the herbal cake-partitioned moxibustion group were treated with additional herbal cake-partitioned moxibustion group at Back-Shu Points and Jiaji (EX-B2) Points once a day. Those in the flour cake-partitioned moxibustion group were treated with the same methods as in the herbal cake-partitioned moxibustion group, except that the herbal cake was replaced by a flour cake. All the patients were treated for 10 d. After treatment, the scores of the visual analog scale (VAS) and Japanese Orthopaedic Association (JOA) and the changes of the interleukin (IL)-6, tumor necrosis factor (TNF)-α, and substance P (SP) levels were observed, and the efficacy was evaluated. Results: After treatment, the VAS score and the levels of IL-6, TNF-α, and SP were lower than those before treatment, and the JOA score was higher than that before treatment in the three groups, indicating intra-group statistical significance (P<0.05). The VAS score and the levels of IL-6, TNF-α, and SP of the herbal cake-partitioned moxibustion group were lower than those of the flour cake-partitioned moxibustion group and the Western medication group, while the JOA score of the herbal cake-partitioned moxibustion group was higher than that of the other two groups, indicating inter-group statistical significance (P<0.05). The total effective rate of the herbal cake-partitioned moxibustion group was 92.5％, higher than that of the flour cake-partitioned moxibustion group (80.0％) and the Western medication group (72.5％), indicating inter-group statistical significance (P<0.05). Conclusion: On the basis of Western oral medication, additional herbal cake-partitioned moxibustion can alleviate the pain and improve the lumbar functions in patients with LDH due to kidney deficiency and blood stasis. The efficacy of the integrated method is better than that of either flour cake-partitioned moxibustion or Western medication alone, which may be related to the reduction of serum inflammatory factors.

OBJECTIVETo explore safe, effective, simple and easy non-drug treatments for grade 1 essential hypertension.

METHODSAccording to TCM syndrome differentiation, 126 cases of grade 1 essential hypertension were classified into 4 types: liver-fire hyperactivity syndrome, yin-deficiency and yang-hyperactivity syndrome, excessive phlegm-dampness syndrome, yin-yang deficiency syndrome, and then the patients were randomly divided into a photoelectric combined with musical treatment group (group A), an acupuncture group(group B) and a placebo group (group C). The acupoints were selected according to TCM syndrome differentiation in group A and group B, and multi-mode audio frequency pulse photoelectric therapeutic apparatus and acupuncture were used in the two groups respectively, once daily. Taichong (LR 3) and Quchi (LI 11) were selected in liver-fire hyperactivity syndrome, Taixi (KI 3) and Sanyinjiao (SP 6) were selected yi yin-deficiency and yang-hyperactivity syndrome, Zusanli (ST 36) and Fenglong (ST 40) were selected in excessive phlegm-dampness syndrome, while Taixi (KI 3) and Guanyuan (CV 4) were selected yi yirryang deficiency syndrome. The group C was treated with oral administration of starch tablet (25 mg), one tablet each time,three times everyday. Ten days were considered as one course, totally three courses were required in the three groups. The blood pressure and scores of TCM syndromes before and after treatment were compared among the three groups.

RESULTSThe blood pressure decreased significantly after treatment in group A and group B (all P<0.01), and the decrease in systolic blood pressure was more significant in group A (P < 0.05). The total effective rate was 90.5 / (38/42) in group A, which was superior to 71. 4 (30/42, P < 0.05) in group B and 19.1% (18/34, P<0. 01) in group C. The scores of TCM syndromes were both improved in group A and group B, but without significant difference between the two groups (P > 0.05).

CONCLUSIONThe clinical effect of multi-mode audio frequency pulse photoelectric therapeutic apparatus for treatment of grade 1 essential hypertension is reliable. Meanwhile, it has the advantages of a non-invasive and simple operation.

Acupuncture Points ; Adult ; Aged ; Blood Pressure ; Combined Modality Therapy ; Electric Stimulation Therapy ; Essential Hypertension ; Female ; Humans ; Hypertension ; physiopathology ; therapy ; Laser Therapy ; Male ; Middle Aged ; Music Therapy ; Treatment Outcome

OBJECTIVETo establish a simple and practical method for screening of Env-specific monoclonal antibodies from HIV-1 infected individuals.

METHODSHuman B cells were purified by negative sorting from PBMCs and memory B cells were further enriched using anti-CD27 microbeads. Gp120 antigen labbled with biotin was incubated with memory B cells to specifically bind IgG on cells membrane. The memory B cells expressing the Env-specific antibody were harvested by magnetic beads separating, counted and diluted to the level of single cell in each PCR well that loading with catch buffer containing RNase inhibitor to get RNAs. The antibody genes were amplified by single cell RT-PCR and nested PCR, cloned into eukaryotic expression vectors and transfected into 293T cells. The binding activity of recombinant antibodies to Env were tested by ELISA.

RESULTSThree monocolonal Env-specific antibodies were isolated from one HIV-1 infected individual.

CONCLUSIONWe can obtain Env-specific antibody by biotin labbled antigen, magnetic beads separating technique coupled with single cell RT-PCR and expression cloning.

Amino Acid Sequence ; Antibodies, Monoclonal ; isolation & purification ; B-Lymphocytes ; immunology ; HIV Antibodies ; isolation & purification ; HIV Envelope Protein gp120 ; immunology ; HIV-1 ; immunology ; Immunomagnetic Separation ; methods ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; methods

Dermatomyofibroma is a benign and rare proliferation of myofibroblasts and fibroblasts of the skin.Dermatomyofibroma commonly locates at the shoulder and neck of young adults and adolescents.Other frequently affected anatomic sites are upper arms,thigh,chest wall,back,axillary region and abdomen.Herein,we reported a case of dermatomyofibroma occurred in the nasion.The asymptomatic firm nodule and histopathological features were consistent with dermatomyofibroma.Immunohistochemically,the tumor cells expressed vimentin,HHF35 and α-smooth muscle actin (α-SMA).The patient was followed up for 2 years after excision of the tumors and recurrences were not observed.

Objective To evaluate the influence and significance of preoperative design of surgical approach on stem cell transplantation via stereotactic surgery. Methods Six patients with stroke in the basal ganglia region were selected. The transplantation target and transcranial approach point were designed by magnetic resonance examination before stem cell transplantation via stereotacfic surgery to guarantee that the line connecting the transplantation target and transcranial approach point could avoid the important functional areas, the ventricular system and the softening focus. Postoperative magnetic resonance examination was performed to observe whether the practical target and surgical approach coincided with the preoperative design or not. Results The practical transplantation target was coincided with the designed transplantation target, distributed around the softening focus without implanted cells in the softening focus. Surgical approach was coincided with the preoperative design and it successfully avoided the important brain functional area, ventricular system and softening focus.Conelnsion The preoperative design of surgical approach can not only ensure the cells being exactly transplanted into the reservation target and guarantee the curative effect, but also promise the surgical approach successfully avoiding the important brain functional area, ventricular system and softening focus and reduce the operative injury.

OBJECTIVETo explore the relationship and interaction of elevated fasting glucose and hypertension on cardiocerebral vascular disease.

METHODS10 054 males and females were recruited for our cross-sectional study during May 2007 to August 2007. Unconditional logistic regression was used to analysis the relationship between fasting glucose and hypertension on cardiocerebral vascular disease. A product of fasting glucose and hypertension was added to the logistic regression model to evaluate the multiplicative interaction and relative excess risk of interaction (RERI), attributable proportion (AP) of interaction and synergy index (S) was applied to evaluate the additive interaction of the two factors. Bootstrap was used to calculate 95% confidence intervals (CI) of RERI, AP and S.

RESULTSAfter adjusting age, gender, smoking, drinking, body mass index (BMI) and region, the product of fasting glucose and hypertension was not statistically significant, which means there was no multiplicative interaction between the two. But the additive indexes RERI, AP and S with 95%CI of diabetes and hypertension were 0.64 (0.03, 1.25), 0.27 (0.01, 0.47) and 1.83 (1.02, 5.13) respectively, which means significant additive interaction was shown between the two on cardiovascular disease but not no stroke. And there were no additive interaction between impaired fasting glucose on cardiovascular disease or stroke.

CONCLUSIONSHypertension was independently related to cardiovascular disease and stroke in Beijing citizens, and diabetes were independently related to stroke. There was additive interaction between diabetes and hypertension on cardiovascular disease.

Adult ; Aged ; Blood Glucose ; metabolism ; Blood Pressure ; Cardiovascular Diseases ; epidemiology ; Cerebrovascular Disorders ; epidemiology ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Hypertension ; blood ; complications ; Male ; Middle Aged ; Risk Factors

OBJECTIVETo investigate the feasibility and clinical effects of titanium elastic nails (TEN) for adolescent clavicular fracture.

METHODSFrom October 2008 to November 2009, 17 adolescent patients with clavicular fracture were treated with internal fixation, including 11 males and 5 females who ranged in age from 12 to 18 years (mean 15.3 years). The mean time from injury to surgery was 3.5 days (2-7 days). Constant function score before surgery and that 3 months after surgery and shoulder ROM before surgery and that 2 months after surgery were compared. Fracture reduction and healing were followed up by X-rays to analyze internal fixation with the TEN technique.

RESULTSAll patients were followed up for a mean of 6.5 months (range 3-8 months), during which no infection, TEN fracture or skin bursting was observed. The Constant score rose from preoperative (45.3 +/- 6.1) to (85.6 +/- 4.3) at 3-month follow up (t = 22.164, P < 0.01),and the shoulder activity degree at 2-week follow up was improved obviosly from preoperative (P < 0.01). X-ray at 12-16 weeks after surgery showed good bone healing and recovery of shoulder function.

CONCLUSIONInternal fixation with TEN in the treatment of adolescent clavicular fracture is safe, minimally invasive,reliable and cosmetic. This technique provides a liable ption for the treatment of adolescent clavicular fractire

Adolescent ; Bone Nails ; Child ; Clavicle ; injuries ; surgery ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Range of Motion, Articular

OBJECTIVETo explore the effect of recombinant human erythropoietin (rhEPO) on expression of brain-derived neurotrophic factor (BDNF) in different brain regions of aging rats.

METHODSForty male SD rats were randomized equally into negative control group, D-galactose group, EPO treatment group, and positive control group. Rat models of subacute aging were established by continuous subcutaneous injection of 5% D-galactose. Immunohistochemical staining was used to analyze the variation of BDNF expressions in different brain regions of the aging rats with different treatments.

RESULTSSignificant brain region-specific differences in BDNF expression were found among the rats in different groups. Compared with those in the negative control group, the numbers of BDNF-positive cells in the hippocampal CA1 region, CA3 region, dentate gyrus (DG) and frontal cortex were all decreased obviously in D-galactose group (P<0.05) but increased in both EPO group and the positive control group (P<0.05) without significant differences between the latter two groups. In the rats in the same group, the number of BDNF-positive cells varied markedly in different brain regions (P<0.05), and the expression level of BDNF was the highest in the frontal cortex followed by the hippocampal CA3 region and the dentate gyrus, and was the lowest in the hippocampal CA1 region.

CONCLUSIONTreatment with rhEPO enhances the expression of BDNF in rat neural cells, suggesting that rhEPO may protect the nervous system from aging by regulating the BDNF pathway.

Aging ; Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; CA1 Region, Hippocampal ; metabolism ; CA3 Region, Hippocampal ; metabolism ; Dentate Gyrus ; metabolism ; Erythropoietin ; pharmacology ; Frontal Lobe ; metabolism ; Galactose ; Humans ; Male ; Neurons ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; pharmacology

OBJECTIVETo investigate the significance of mutation and single nucleotide polymorphism (SNP) of class III receptor tyrosine kinases such as PDGFRbeta and SHIP in acute myeloid leukemia (AML) patients.

METHODSScreening of the mutation and SNP of PDGFRbeta and SHIP by genomic PCR, RT-PCR, directly sequencing and Mass-ARRAY system was carried out in 273 AML patients.

RESULTSThe mutations of PDGFRbeta R685C and SHIP Q1153L were detected for the first time in AML patients. The positivity ratio was 0.73% and 0.36% respectively.

CONCLUSIONThe mutations of PDGFRbeta R685C and SHIP Q1153L may contribute to leukemogenesis of AML.

Humans ; Inositol Phosphates ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Mass Spectrometry ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Receptor, Platelet-Derived Growth Factor beta ; genetics ; Reverse Transcriptase Polymerase Chain Reaction