Objective To observe the protective effect of levo-anlodipine on the calcium overload in the anoxic myocardial cells. Methods Myocardial cells were cultured, and in vitro model of anoxic myocardial cells was established. Fura2/am fluorescent probe was used to measure the free calcium concentration of the myocardial cells by spectrofluorometer. The expression of Ca~2+ ATPase of cellular membrane was detected by RT-PCR, and the content of calcium pump in sarcoplasmic reticulum was measured by western blot. Results The levo-anlodipine could decrease the free calcium concentration of the anoxic myocardial cells markedly(359.06?75.00nM), and increase the expression of Ca~2+ ATPase of cellular membrane and the content of calcium pump of sarcoplasmic reticulum in the anoxic myocardial cells. Conclusion Levo-anlodipine could enhance the resistance to calcium overload in anoxic myocardial cells through increasing the expression of Ca~2+ ATPase.

Objective To observe the impact of lysophosphatidyl choline(LPC) on the expression of scavenger receptor class B type Ⅰ(SRBI) in L-20 cells and the impact of simvastatin.Methods Cultured hepatocytes were randomly assigned to normal group,LPC-damaged group and simvastatin group.The changes of mRNA and protein expression of SRBI in hepatic tissue were assayed by RT-PCR,immunocytochemistry and Western blot,respectively.Results The expression of SRBI was higher in LPC-damaged hepatocytes than that in normal cells at both mRNA and protein level(P

Traditional right ventricular pacing is non-physiological pacing, can lead to ventricular electrical and mechanical dyssynchmnization, which increases the risk of heart failure and atrial fibrillation. His-Purkinje system pacing is the hot research of physiological pacing method. The high thresholds and difficult implantation of his bundle pacing limits its application and popularization. In recent years, domestic scholars have proposed left bundle branch area pacing. Many studies have confirmed the safety and feasibility of left bundle branch area pacing. Left bundle branch pacing transmit down the conduction system, which can effectively achieve ventricular electrical and mechanical synchrony, At the same time, it also has many others advantages, such as good and stable electrical parameters, significantly improved cardiac function, and pacing across blocked parts. It will become the main development direction of physiological pacing in the future. This article aims to review the origin, standard and classification, advantages and limitations of left bundle branch area pacing.

AIM: To observe the influence of serum contained Tongmai herbal medicine on calcium overload of the anoxic myocardial cells. METHODS: Serum contained Tongmai herbal medicine was prepared with the serum pharmacological method and the model of anoxic myocardial cells was established. The intracellular free calcium concentration and the expression of the L-type calcium channel were determined with Fura-2/AM and RT-PCR, respectively. RESULTS: The serum contained Tongmai herbal medicine and levo-anlodipine decreased the intracellular free calcium concentration in the anoxic myocardial cells. The serum contained Tongmai herbal medicine also suppressed the expression of L-type calcium channel in the anoxic myocardial cells, but levo-anlodipine had no effect on the expression of L-type calcium channel. CONCLUSION: Tongmai herbal medicine can attenuate calcium overload in anoxic myocardial cells.

[Objective] To investigate combined application of multiplex reverse transcription-polymerase chain reaction(mRT-PCR)and karyotype analysis detect of clonal chromosomal aberrations in acute lymphoblasfic leukemia (ALL),and explore the expression of common fusion genes.Methods 189 ALL patients were examined by multiplex RT-PCR and R or G banding techniques.[Results]10 fusion genes were detected in 69 out of 189 ALL patients(36.5％),including E2A/PBX1,TEL/AML1,BCR/ABL,MLL/AF4,MLL/AF6、MLL/AF9,MLL/AF10,MLL/ELL,SIL/TAL1,TLS/ERG.R or G banding techniques could find chromosome structural and numeracy abnormalities in 86 out of 152 patients (56.6％) available for analysis.Combination of mRT-PCR and R or G banding could raise the rate of detecting clonal chromosomal abnormalities to 69.3％.Fusion genes were detected in 33 out of 90 (36.7％) patients with adult ALL and 36out of 99 (36.4 ％) patients with children ALL,there were 22 patients with positive BCR/ABL but no TEL/AML1 in adult ALL group,while there were 24 patients with positive TEL/AML.1 and 2 with positive BCR/ABL in children ALL group.There was significant statistical difference for the expression of RCR/ABL and TEL/AML1 between adult ALL and children ALL (P＜0.01),but no difference for MLL related fusion gene,E2A/PBX1,SIL/TAL1 and TLS/ERG(P＞0.05).BCR/ABL and TEL/AML1 fusion gene could be detected in 66 ALL patients with normal karyotype (36.3％).[Conclusion]There were different biological characteristics between adults and children with ALL.mRT-PCR technique can quickly screen chromosome structural aberrations in patients with newly diagnosed leukemia.It is useful in detection of fusion genes in ALL with normal karyotypes and it would refine the karyotype analysis and provide imrootramt prognosis-relevant information.

Objective To explore the value of fluorescence in situ hybridization (FISH) and multiplex RT-PCR in the detection of mixed lineage leukemia (MLL) gene rearrangement in acute leukemia (AL) patients. Methods Dual-color MLL probe, multiplex RT-PCR and R or G banding techniques were used to detect the MLL gene rearrangement in 189 cases of AL.Results MLL gene rearrangements were detected in 9 cases (5.03 ％) by FISH,and 16 cases (8.47 ％) by multiplex RT-PCR,including MLL/AF9,MLL/AF10,MLL/AF6, MLL/AF7, MLL/ELL, MLL/PTD. R or G banding techniques could find 11q23 in 5 out of 189 patients (2.65 ％). There was no statistic difference in the incidence of 6 common MLL gene rearrangements between ALL (73 cases) and AML patients (116 cases) (P ＞ 0.05).Conclusion Multiplex RT-PCR is a powerful technique in the detection of MLL gene rearrangement for tentatively diagnosed AL.It could not only confirm translocation detected by conventional cytogenetic method, but also detect MLL partial tandem duplication which could not been detected by cytogenetic examination or FISH. It plays an important role in guiding therapy and predicting prognosis for AL.

BACKGROUND:Calcium channel abnormalities can damage myocardium. Heroin can directly affect calcium ion channel, and alter myocardial structure.
OBJECTIVE:To study the changes of heroin-induced myocardial ultrastructure, L-type calcium channel current and action potential of myocardial cel s after rat cardiac arrhythmia.
METHODS:Sprague-Dawley rats were randomly divided into control group and model group. In the model group, rats were administered heroin at initial dose of 5 mg/kg?d. The daily dose escalation method was used (increasing dose:2.5 mg/kg?d) to replicate rat models of heroin addiction. At 20 days, models of heroin addiction were successful y established. At 30 days after increasing the dose, rat models of heroin addiction-induced arrhythmias were further established.
RESULTS AND CONCLUSION:Compared with the control group, the electron microscopy demonstrated that myocardial structure changes mainly presented nuclear membrane shrinkage, nuclear condensation, nuclei became smal , chromatin assembled into blocks, mitochondria disordered and disappeared, sarcomeres disordered, focal fracture, and unclear myofilament in rat models of heroin addiction. Electric current-voltage curve of myocardial cel L-type calcium channel current showed the increasing trends. The 90%repolarization action potential was significantly shortened. These data indicated that heroin can directly lead to the pathological change of myocardial structure. Calcium channel current change is one of the main reasons for myocardial injury.

Abstract BACKGROUND:Caspase-8 plays an important role in starting the process of cel apoptosis, and diacetylmorphine can induce neuronal apoptosis. The relationship between the apoptosis of cerebelar granule neurons cels induced by diacetylmorphine and caspase-8 has not been reported. OBJECTIVE:To investigate the effect of caspase-8 on diacetylmorphine-induced neuronal apoptosis. METHODS:Cerebelar granule neurons from Sprague-Dawley rats aged 7 days were culturedin vitro. At 7 days, the cels were cultured with different dosage of diacetylmorphine (10, 40, 80, 100, 120 mg/L) and SP600125 for 24 hours, and were divided into control group, 80 mg/L diacetylmorphine group, diacetylmorphine+SP600125 group. Cel morphology was observed by Hoechst 33258 fluorescent staining, and cel apoptosis rate was measured by flow cytometry. Immunofluorescence staining, RT-PCR and western blot assay were used to detect caspase-8 mRNA and protein expression. RESULTS AND CONCLUSION:(1) After different dosage of diacetylmorphine was used to induce neuronal apoptosis, dark blue translucent apoptotic bodies were found in apoptotic cels, appearing with nucleus condensation, cohesion and fracture, and the apoptosis rate presented an increasing trend with increasing of drug dosage (P < 0.05). (2) Compared with the control group, caspase-8 mRNA and protein expression was remarkable under the intervention of 80 mg/L diacetylmorphine(P < 0.05); caspase-8 mRNA expression exhibited an increasing trend with increasing dosage of diacetylmorphine (P < 0.05), while caspase-8 mRNA and protein expression in the diacetylmorphine+SP600125 group was significantly lower than that in the 80 mg/L diacetylmorphine group (P < 0.05). These findings indicate that caspase-8 is involved in the process of diacetylmorphine-induced neuronal apoptosis, and meanwhile, it is also an important candidate of pro-apoptotic factors in the c-jun N-terminal kinase signaling pathway.

Objective To study the effect and safety of GDP regimens (gemcitabine,cisplatin,dexamethasone) as salvage chemotherapy in relapsed refractory non-Hodgkin’s lymphoma (NHL).Methods Clinical response and adverse effects of 34 NHL patients treated by GDP regimens were analyzed retrospectively.Results The overall response rate (ORR) was 52.9 ％ (18/34),there was no statistic difference between ORR in relapsed NHL [72.7 ％ (8/11)] and that in refractory ones [43.5 ％ (10/23)](P =0.11).The ORR for B-NHL was 64.0 ％ (16/25),while for T-NHL was 22.2 ％ (2/9),there was no statistic difference between them (P =0.052).The ORR for low risks groups was 66.7 ％ (6/9),while it for intermediate-risk and high-risk ones were 55.6 ％ (10/18) and 28.6 ％ (2/7),respectively,the effect of GDP regimens for low-risk group was better for high-risk ones,but there was no statistic difference (P =0.349).The incedience of Ⅲ-Ⅳ granulopenia reduced was 23.5 ％ (8/34) and there were no Ⅲ-Ⅳ gastrointestinal adverse reactions or liver and kidney toxicity.Conclusion GDP regimens were efficient and less toxic as second-line salvage chemotherapy.

OBJECTIVE:To establish the method for simultaneous determination of the content of berberine hydrochloride, baicalin and osthole in Jinchan zhiyang capsules. METHODS:HPLC method was adopted. The determination was performed in Hypersil BDS C18 column with mobile phase consisted of methanol-acetonitrile-0.1%phosphoric acid-three triethylamine(50∶30∶19∶1, V/V/V/V) at the flow rate of 1.0 mL/min. The detection wavelengths were set at 265 nm (berberine hydrochloride),280 nm (baicalin)and 322 nm(osthole). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride,baicalin and osthole were 80-800 ng(r＝0.999 8),60-600 ng(r＝0.999 9),60-600 ng(r＝0.999 6),respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The limits of quantitation were 80,60,60 ng,respectively,and the limits of detection were 24,20,20 ng,respectively. The recovery rates were 97.4%-98.3%(RSD＝0.33%,n＝6),98.4%-99.6%(RSD＝0.42%,n＝6)and 96.9%-99.0%(RSD＝0.92%,n＝6),respectively. RSDs of durability tests were all lower than 1.2%. CONCLUSIONS:The method is simple, accurate, precise, stable, reproducible and durable. It can be used for simultaneous determination of berberine hydrochloride,baicalin and osthole in Jinchan zhiyang capsules.