Objective:To study the effect of electroacupuncture (EA) at the Pericardium Meridian on the heart function of volunteers with acute hypoxia, and to provide scientific evidence for the acupoints selection along the affected meridian in acupuncture-moxibustion therapy. Methods:Based on a self-control design, eighteen healthy volunteers were recruited in the study. Points from the Pericardium Meridian [Neiguan (PC 6), Ximen (PC 4), Quze (PC 3) and Tianquan (PC 2)], non-Pericardium Meridian point [Shousanli (LI 10)], non-meridian and non-acupoint points [1.0-1.5 cm lateral to Neiguan (PC 6) and Ximen (PC 4), respectively on both sides], and a blank control (only inhaling low-oxygen gas without EA stimulation) were selected to observe, once every week, 10 sessions in total, and only 1 acupoint was observed once. The volunteers inhaled low-oxygen gas mixture (10.8% O2 and 89.2% N2) for 30 min to imitate acute hypoxia. EA was conducted when the gas mixture was inhaled for 10 min and then lasted for 20 min; meanwhile, hemodynamic indexes such as cardiac output (CO), cardiac index (CI), systemic vascular resistance (SVR), systemic vascular resistance index (SVRI), left cardiac work (LCW), left cardiac work index (LCWI) and heart rate (HR) were recorded on a hemodynamic monitor. Results:EA at the acupoints of Pericardium Meridian significantly down-regulated the increased CO/CI, LCW/LCWI, and HR (P<0.05), and significantly up-regulated the decreased SVR/SVRI in hypoxia (P<0.05); EA at other meridian acupoints or at non-meridian and non-acupoint points didn't produce such effects. Conclusion: EA at the Pericardium Meridian can obviously improve the cardiac hyper-activation caused by acute hypoxia in healthy volunteers.

Nowadays, the 5-year survival rate of childhood cancer patients can be more than 80%, but some patients with relapse and refractory cancers have shown no good response to traditional strategies. Chimeric antigen receptor engineered T (CAR-T) cell therapy is promising for these patients. CAR-T cells recognize the tumor-associated antigens in a non-major histocompatibility complex-restricted manner, so their anti-tumor ability is enhanced. There are four generations of CAR-T cells now. The complete remission rate of pediatric patients with relapse and refractory acute lymphoblastic leukemia can be as high as 90% when treated with CD19-targeting CAR-T cells. Furthermore, CAR-T cell therapy can also be used to bridge to transplantation and donor CAR-T cell infusion can be a strategy to prevent relapse after hematopoietic stem cell transplantation. As to solid tumors, only patients with neuroblastoma present good response to the GD2-targeting CAR-T cell therapy. The toxic or side effects of CAR-T cell therapy include cytokine release syndrome, off-tumor effect, tumor lysis syndrome, and insertion mutation. Although the CD19-targeting CAR-T cell therapy for childhood cancer can result in a high remission rate, the relapse rate is high, including CD19and CD19relapse. The mechanisms for relapse merit further investigatio.
Antigens, CD19 ; immunology ; Child ; Humans ; Immunotherapy, Adoptive ; adverse effects ; methods ; Neoplasms ; therapy ; Receptors, Antigen, T-Cell ; genetics ; T-Lymphocytes ; transplantation

OBJECTIVECigarette smoke extract (CSE) can induce injuries of pulmonary II epithelial cells, activate nuclear factor-kappaB and increase tumor necrosis factor-alpha(TNF-alpha) secretion. This study aimed to investigate whether azithromycin can protect pulmonary II epithelial cells from injuries induced by CSE and relevant mechanisms.

METHODSPulmonary II epithelial cells (A549 cells) were cultured in vitro. After 48 hrs of culture the cells were randomly treated with serum-free DMEM only (blank control group), azithromycin + serum-free DMEM, CSE+ serum-free DMEM or CSE+azithromycin. Eight hours later the morphology of A549 cells, the activity of NF-kappaB and the levels of TNF-alpha were measured by inverted microscope, immunohistochemistry and ELISA.

RESULTSThe morphology and structure of A549 cells were changed, NF-kappaB activity increased (dark brown staining ) and TNF-alpha levels (0.307 +/- 0.036 pg/mL vs 0.234 +/- 0.028 pg/mL)increased in the CSE+ serum-free DMEM group compared with the blank control group (P < 0.01). CSE together with azithromycin treatment recovered partly the morphological injuries of A549 cells. It also attenuated NF-kappaB staining and decreased TNF-alpha levels from 0.307 +/- 0.036 pg/mL (CSE+serum-free DMEM group) to 0.269 +/- 0.009 pg/mL (P < 0.05).

CONCLUSIONSAzithromycin may inhibit NF-kappaB activity, decrease TNF-alpha secretion and thus lessen cytotoxicity of CSE to A549 cells.

Anti-Bacterial Agents ; pharmacology ; Azithromycin ; pharmacology ; Cells, Cultured ; Epithelial Cells ; drug effects ; Humans ; Immunohistochemistry ; Lung ; drug effects ; metabolism ; pathology ; NF-kappa B ; analysis ; Smoke ; adverse effects ; Tobacco ; adverse effects ; Tumor Necrosis Factor-alpha ; analysis

Adult ; Female ; Humans ; Middle Aged ; Myxoma ; pathology ; surgery ; Vulvar Neoplasms ; pathology ; surgery

Objective To establish a kind of animal model and methods for neuroendoscope training. Methods With rat abdominal cavity hypothesized as cerebral ventricle, a set of programs for neuroendoscopic operative skill training were designed, including endoscopic techiniques, electronic coagulation, suction, flush, biopsy and balloon dilatation, and so on. Results Simulation operation was performed on rat abdomen under neuroendoscope. The procedure helped to practice the endoscopic manipulation, get to know well how to perform endoscope, coagulation, suction, flash instruments, be familiar with the usage of electric coagulation in the liquid condition. The emphasis was put on the basic skills of pure neuroendoscopic operation such as balloon dilatation, electric coagulation, cutting off,forcep biopsy and flush. Conclusions The rat model can provide a way to train pure neuroendoscopic operation. The trainer can get basic experience with regard to endoscopic manipulation, balloon dilatation, electric coagulation, suction, flush and biopsy under endoscope. The method can be an important part of neuroendoscopic laboratory training.

Objective To investigate the neuroprotective effects of angiotensin Ⅱ type 1 receptor (AT1R) blocker olmesartan medoxomil (OLM) on intracerebral hemorrhage (ICH) in spontaneously hypertensive rats (SHRs). Methods SHRs (male, 12 weeks old; weighing 300±20 g) were randomly assigned to normal, ICH, vehicle-treatment ICH (control), OLM-treatment ICH (OLM) groups. ICH was induced via stereotaxic right basal ganglia administration of collagenase type Ⅶ. One hour after ICH, the rats in OLM group were given a single oral dose of OLM (10 or 3 mg/kg solved in 1 mL sodium carboxymethylcellulose) via nasogastric feeding, and those in the control group received an equal volume of sodium carboxymethylcellulose only. Six hours after ICH induction, mean arterial blood pressure (MAP) was measured using the non-invasive method of tail-cuff plethysmography in conscious rats. Twenty-four hours after ICH induction, neurobehavior was detected by the modified limb placing test (MLPT); brain water content was measured by dry-wet method; the mRNA expression levels of receptor and target genes were analyzed by real-time PCR. Results MAP in the ICH group ([121.4±3.5] mm Hg) did not significantly differ from baseline pressure in the normal group ([120.2±3.8] mm Hg)(P>0.05); MAP in the OLM group with 10 mg/kg ([105.6±3.1] mm Hg) was significantly lower than that in the ICH group (P<0.05); the OLM group with 3 mg/kg ([120.8±3.1] mm Hg) and control group ([118.6±3.9] mm Hg) did not induce blood pressure reduction, and did not show significant difference as compared with the ICH group (P>0.05). In the hemorrhagic hemisphere, brain water content in the OLM group with 3 mg/kg (80.02%±0.32%) had significant difference from that in the ICH group (80.90%±0.36%, P< 0.05); brain water content of the control group (80.81%±0.32%) was slightly lower than that of the ICH group, without significant differences (P>0.05). The OLM group with 3 mg/kg (5.03±0.71) was showed significantly lower score of MLPT as compared with that in the ICH group (6.62±0.55, P<0.05). The score of MLPT in the control group (6.41 ±0.55) did not differ from that in the ICH group (P>0.05). In the hemorrhagic hemisphere, the mRNA expressions of AT1R and target genes, such as HO-1, COX-2, IL-6 and VCAM-1, in the OLM group with 3 mg/kg were significantly lower than those in the ICH group (P<0.05), but the difference between the control and ICH groups did not show statistical significance (P>0.05). Conclusion Treatment with low doses of OLM in the experimental ICH of SHRs may promote its neurological recovery and induce its neuroprotective effects, including reduction of edema, inhibition of inflammation and oxidative stress.

OBJECTIVETo investigate the methylation status of the promoter of resion death associated protein kinase (DAPK) gene in bladder cancer cell (T24), and study the effect of 5-aza-2'-deoxycytidine (5-aza-dc) on DAPK gene reactive expression in T24 and its inhibitory effect on T24.

METHODSThe bladder cancer cell T24 was treated with different doses of 5-aza-dc. The inhibitory effect and apoptosis rate were detected by MTT and flow cytometry, and the changes of DAPK mRNA and protein expression and the methylation status of DAPK promoter were assessed by RT-PCR, Western blotting, and methylation specific PCR, respectively.

RESULTSThe growth of bladder cancer cell was inhibited significantly and the maximal apoptosis rate detected by flow cytometry was (24.12-/+1.4)%. DAPK mRNA was not expressed in bladder cancer cell T24 in normal conditions. DAPK mRNA and protein re-expressed after 5-aza-dc (12.5 micromol/L) treatment in cell line T24 for 24 h, and DAPK promoter became unmethylated.

CONCLUSIONSThe promoter methylation can be an important factor for silencing the expression of DAPK in bladder cancer cell. 5-aza-dc can inhibit the growth and induce apoptosis of bladder cancer cells through reversing unmethylation status of DAPK promoter.

Apoptosis Regulatory Proteins ; genetics ; metabolism ; Azacitidine ; analogs & derivatives ; pharmacology ; Calcium-Calmodulin-Dependent Protein Kinases ; genetics ; metabolism ; Cell Line, Tumor ; DNA Methylation ; DNA Modification Methylases ; antagonists & inhibitors ; Death-Associated Protein Kinases ; Humans ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; metabolism ; Transcriptional Activation ; drug effects ; Urinary Bladder Neoplasms ; metabolism ; pathology

OBJECTIVETo understand the characteristics of embB gene mutation of Mycobacterium tuberculosis (MTB) isolates from tuberculosis patients in Chongqing, and the value of embB306 as a molecular marker used to diagnose ethambutol (EMB)-resistant MTB strains.

METHODSDirect sequencing was used to analyze the polymorphism of embB mutation in 51 EMB-resistant MTB strains and 50 EMB-sensitive MTB strains. And diagnostic testing was used to evaluate the value of embB306 as a molecular marker of EMB -resistant MTB strains as compared with the traditional sensitivity test.

RESULTSAll 34 of 51 EMB-resistant strains (66.7%) and 3 of 51 EMB-sensitive strains (6%) had had embB306 mutation. The embB306 mutation rate in EMB-resistant strains coming from previously treated case was 87.5%, showing significantly higher than that from new cases (48.1%, P < 0.01); embB306 mutation rate was increased with the number of the resistant drugs; embB306 mutation serving as a marker to diagnose EMB-resistant MTB strains comparing with the traditional sensitivity test, had the rate of sensitivity = 66.7%, specificity = 94.0%, accuracy = 80.2% and Youden index = 60.7%.

CONCLUSIONembB306 mutation should be the main mechanism of MTB resistance to EMB in Chongqing, showing an association with the history of the treated and numbers of the resistant drugs. embB306 mutation should be a good marker to diagnose EMB-resistant MTB strains.

China ; DNA Mutational Analysis ; DNA, Bacterial ; genetics ; Genes, Bacterial ; Humans ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; isolation & purification ; Pentosyltransferases ; genetics ; Tuberculosis, Multidrug-Resistant ; microbiology

This study was aimed to investigate the effect of recombinant mutant human TNF-related apoptosis-inducing ligand (rmhTRAIL) combined with As(2)O(3) on inducing apoptosis of adriamycin-resistant leukemia cell line K562/A02 (mdr-1(+)). The morphologic changes of cells treated with rmhTRAIL were observed by inverted microscope, taking adriamycin-sensitive cell line K562 (mdr-1(-)) as control; the inhibitory rate of cell proliferation after being treated with rmhTRAIL, As(2)O(3) alone or combined was assayed by MTT method; the apoptosis peaks of K562/AO2 and K562 were quantitatively detected by flow cytometry with PI staining after being treated with rmhTRAIL, As(2)O(3) alone or in combination. The results indicated that the inhibition effect of rmhTRAIL and As(2)O(3) in combination on K562/AO2 and K562 cells was higher than that of riTRAIL and As(2)O(3) alone (p < 0.01), rmhTRAIL combined with As(2)O(3) had synergistic effect in killing K562/AO2 and K562 cells by king's formula. The apoptosis rates of K562/AO2 and K562 cells were 34.93 +/- 0.10% and 10.53 +/- 0.16% (p < 0.01), as well as 5.95 +/- 0.07%, and 3.50 +/- 0.01% (p < 0.05), 50.95 +/- 0.91% and 20.75 +/- 0.95% (p < 0.05) respectively when their cells were treated by rmhTRAIL and As(2)O(3) alone. The apoptosis rate in K562/AO2 group was higher than that in K562 group. It is concluded that rmhTRAIL can induce K562/A02 and K562 cell apoptosis; rmhTRAIL combined As(2)O(3) had synergistic effects; the efficacy of on rmhTRAIL or As(2)O(3) inducing K562/AO2 cell apoptosis is higher than that on their parental cell line K562.
Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Humans ; K562 Cells ; Oxides ; pharmacology ; Recombinant Proteins ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology

Objective In vitro isolation of adenohypophyseal mainly includes enzymatic digestion and mechanical methods.But there is no relevant report about which method is better.In this paper,several mainstream methods of the vitro isolation of adenohypophysis cells in rats are compared and the identification of gonadotropin secretory cells is carried out to figure out which preparation method is more convenient and efficient.Methods 48 mature female SD rats were randomly divided into 4 groups:trypsin group (0.25％ trypsin EDTA digestion),Ⅳ-type collagen enzyme group (Ⅳ type collagen enzyme digestion),mechanical separation group (200 mesh cell sieves grinding) and trypsin digestion plus mechanical separation group (0.25％ trypsin EDTA digestion plus 200 mesh cell sieves grinding).12 rats in each group.The effect of these 4 methods was evaluated.Secondly,the primary cells of each group were cultured.We dynamically observed the vitro growth of cells in each group.Finally,the cells were identified by using the immunocytochemi-cal staining technique Results Compared with the mechanical separation group and-ⅣV type collagen enzyme group [(90.2 ± 0.96) ％,(93.32± 1.77)％],Cell viability of trypsin group and trypsin digestion plus mechanical separation group was elevated [(94.11 ± 1.71) ％,(94.92± 1.92) ％] (P＜0.05).Morphological observation:The pituitary ceils prepared by each methods were all round with strong refraction and clear edges and began to partly adhere to the wall after cultured for 16-24 hours.Then most of the cells adhered to wall after 48-72 hours,while the early glandular ceils were still round.But the volume was smaller than before.Also,a small amount of large cells were scattered among them.7 days later,the cells began to become polygonal or spindle shaped and were connected to pieces.Till around 10 days,the fibrosis gradually became obvious;Immunocytochemical staining:It is indicated that FSH positive cells were larger in volume while less in number and scattered in the cytoplasm.And the positive products,showing blue,were located in the cytoplasm;The Enzyme linked immunosorbent assay (ELISA) was used to determine the supematant of cells and confirmed that the cells in vitro still had hormone secretion functions.Conclusion Compared with the single cell preparation method,the trypsin digestion plus mechanical separation method has certain advantages;The anterior pituitary cells vitro cultured are in good health conditions and still have secrete functions.They are tested to be competent for related scientific experiments.