[Leydig cell hyperplasia of testis in children: a clinicopathological study].

Objective: To investigate the clinical, pathological features and differential diagnosis of testicular Leydig cell hyperplasia (LCH) . Methods: Clinical data, histological features, immunohistochemical findings, ultrastructural characteristics and follow-up data were analyzed in three cases of LCH. The cases were collected from 2011 to 2014 at Beijing Children's Hospital. A literature review was performed. Results: Two males (1.8 years and 2.9 years of age) showed isosexual pseudoprecocity with elevated serum testosterone. Imaging study showed bilateral testicular enlargement with multiple small nodules in the parenchyma. Another 13 years-old patient showed male pseudohermaphroditism and cryptorchism. Gross examination showed the bilateral markedly enlarged testis without discrete lesion. Histologically, LCH was seen in both nodular and diffuse patterns without destruction of seminiferous tubules. Adjacent spermatogenesis was noted. Immunohistochemically, the Leydig cells were positive for inhibin, calretinin and Melan A and ultrastructural analysis showed enriched cytoplasmic endoplasmic reticulum. Two cases had followed up for 7 years. One patient was symptom-free and one was stable. Conclusion: LCH is a rare benign condition, which is easily misinterpreted as testicular tumor or non-neoplastic diseases. Clinical presentation, imaging study and pathological evaluation are required for the diagnosis.

Expression of Cx43 and Cx45 in Cardiomyocytes of an Overworked Rat Model.

ABSTRACT: Objective To study the effect of overwork stress response on the expression of connexin 43（Cx43） and connexin 45（Cx45） in cardiomyocytes and on cardiac function. Methods The experimental animals were divided into control group, overworked 1-month group and overworked 2-month group. A overworked rat model was established by forcing swimming of overworked group. The expressions of Cx43 and Cx45 in myocardial tissues of experimental animals were detected by Western blotting, while the corresponding myocardial tissues were stained with hematoxylin-eosin （HE） staining and Masson's staining, then histologically observed. Results Western blotting results showed that, compared with the control group, Cx43 expression in myocardial tissues of overworked rats decreased while Cx45 expression increased. HE staining and Masson's staining results showed that hypertrophy, rupture and interstitial fiber tissue hyperplasia were observed in myocardial fibers of overworked rats. Conclusion Overwork stress response may affect cardiac function as an independent factor and may even cause heart failure or arrhythmias and lead to death.

The Concept, Status Quo and Forensic Pathology of Karoshi.

ABSTRACT: "Karoshi" originates from Japan's economic take-off period in the 1960s and 1970s. It is generally believed that overwork lead to the accumulation of fatigue, which triggers the outbreak of potential diseases, and results in sudden death. Karoshi causes great harm to both the community and families because it occurs primarily in 30 to 60 year old young adults. Japan put Karoshi into the category of industrial injury for the first time in 2001 and started to undertake a series of studies in the sociological and pathological fields. However, there is a tremendous gap in the forensic pathological diagnosis domain. In China, research on Karoshi started from the 1990s and is closely related to the reform and opening up policy as well as economic development. According to the incomplete statistics, 600 thousand people die from overwork each year in China, the highest in the world. Karoshi has become one of the most serious social problems in China at the present stage, thus a systematic study in the sociology and forensic pathology fields is urgently required. This paper summarizes the past and present status of Karoshi, and puts forward the problems that need attention during the judicial expertise of Karoshi from forensic pathology perspective.

Resveratrol protects against arsenic trioxide-induced cardiotoxicity in vitro and in vivo.

BACKGROUND AND PURPOSE: The clinical use of arsenic trioxide (As(2)O(3)), a potent antineoplastic agent, is limited by its severe cardiotoxic effects. QT interval prolongation and apoptosis have been implicated in the cardiotoxicity of As(2)O(3). The present study was designed to evaluate the effects of resveratrol on As(2)O(3)-induced apoptosis and cardiac injury. EXPERIMENTAL APPROACH: In a mouse model of As(2)O(3)-induced cardiomyopathy in vivo, QT intervals and plasma enzyme activities were measured; cardiac tissues were examined histologically and apoptosis assessed. In H9c2 cardiomyocyte cells, viability, apoptosis, generation of reactive oxygen species (ROS) and cellular calcium levels were measured. KEY RESULTS: In the mouse model, resveratrol reduced As(2)O(3)-induced QT interval prolongation and cardiomyocyte injury (apoptosis, myofibrillar loss and vacuolization). In addition, increased lactate dehydrogenase activity and decreased activities of glutathione peroxidase, catalase and superoxide dismutase were observed in the plasma of As(2)O(3)-treated mice; these changes were prevented by pretreatment with resveratrol. In As(2)O(3)-treated H9c2 cardiomyocytes, resveratrol significantly increased cardiomyocyte viability and attenuated cell apoptosis as measured by acridine orange/ethidium bromide staining, TdT-mediated dUTP nick end labelling assay and caspase-3 activity. As(2)O(3)-induced generation of ROS and intracellular calcium mobilization in H9c2 cells was also suppressed by pretreatment with resveratrol. CONCLUSIONS AND IMPLICATIONS: Our results showed that resveratrol significantly attenuated As(2)O(3)-induced QT prolongation, structural abnormalities and oxidative damage in the heart. In H9c2 cardiomyocytes, resveratrol also decreased apoptosis, production of ROS and intracellular calcium mobilization induced by treatment with As(2)O(3). These observations suggested that resveratrol has the potential to protect against cardiotoxicity in As(2)O(3)-exposed patients.

LncRNA SNHG12 regulated the proliferation of gastric carcinoma cell BGC-823 by targeting microRNA-199a/b-5p.

OBJECTIVE: We aimed at detecting the expression of lncRNA SNHG12 in gastric carcinoma and at exploring its effect on the proliferative ability and metastasis of gastric carcinoma. Also, its mechanism was analyzed. PATIENTS AND METHODS: A total of 54 pairs of gastric carcinoma and paracancerous tissues were harvested from patients in our hospital. Clinical data were used for analyzing the relationship between lncRNA SNHG12 and general information of patients with gastric carcinoma. Knockdown and overexpression experiments were carried out to investigate the relationship between the expression of lncRNA SNHG12 and the proliferation ability of BGC-823. RNA levels of possible target microRNAs were verified and further confirmed by RIP and CHIP experiments. RESULTS: Higher lncRNA SNHG12 expression was observed in 6 gastric carcinoma tissues than the paracancerous tissues. Clinical data analysis demonstrated that highly expressed lncRNA SNHG12 was related to tumor size and TNM staging. Survival analysis showed that highly expressed lncRNA SNHG12 in patients with gastric carcinoma was negatively correlated to the overall survival time. Additionally, the expression of lncRNA SNHG12 was positively linked with the proliferative ability of BGC-823. RIP experiment confirmed the binding abilities of lncRNA SNHG12, microRNA-199a/b-5p and Argo2. The distinct interaction between lncRNA SNHG12 and microRNA-199a/b-5p was verified by CHIP experiment CONCLUSIONS: This study indicates that lncRNA SNHG12 may play an important role in tumorigenesis and may serve as a molecular target for the malignant gastric carcinoma.

Advances in exploring the role of microRNAs in the pathogenesis, diagnosis and therapy of cardiac diseases in China.

Cardiovascular disease has become the most serious health threat and represents the major cause of morbidity and mortality in China, as in other industrialized nations. During the past few decades, China's economic boom has tremendously improved people's standard of living but has also changed their lifestyle, increasing the prevalence of cardiovascular disease, the so-called 'disease of modern civilization'. This new trend has attracted a significant amount of research. Many of the studies conducted by Chinese investigators are orientated towards understanding the molecular mechanisms of cardiovascular disease. At the molecular level, the long-standing consensus is that cardiovascular disease is associated with a sequence mutation (genetic anomaly) and expression deregulation (epigenetic disorder) of protein-coding genes. However, new research data have established the non-protein-coding genes microRNAs (miRNAs) as a central regulator of the pathogenesis of cardiac disease and a potential new therapeutic target for cardiovascular disease. These small non-coding RNAs have also been subjected to extensive, rigorous investigations by Chinese researchers. Over the years, a large body of studies on miRNAs in cardiovascular disease has been conducted by Chinese investigators, yielding fruitful research results and a better understanding of miRNAs as a new level of molecular mechanisms for the pathogenesis of cardiac disease. In this review, we briefly summarize the current status of research in the field of miRNAs and cardiovascular disease in China, highlighting the advances made in elucidating the role of miRNAs in various cardiac conditions, including cardiac arrhythmia, myocardial ischaemia, cardiac hypertrophy and heart failure. We have also examined the potential of miRNAs as novel diagnostic biomarkers and therapeutic targets.

Cardiovascular research is thriving in China.

Cardiovascular disease has become the leading cause of death and constitutes a serious public health issue in China. Faced with the burgeoning epidemic of cardiovascular disease and the huge burden and economic losses it causes, the Chinese government has attached the utmost importance to cardiovascular research, increasing funding to support basic and clinical studies, integrating resources and recruiting outstanding talent from overseas. The continued and growing support from the government has yielded substantial changes in terms of new discoveries, scientific publications and drug research and development within the last decade. In spite of the advances in cardiovascular research, China still faces significant challenges ahead in encouraging innovation, developing the prevention-oriented health policies and strengthening international collaboration.

[Inhibition of transmembrane K(+) currents in mammalian ventricular myocytes by antiarrhythmic agent RP62719].

The effect of RP62719 on the inward rectifier K(+) current (I(K1)),transient outward K(+) current (I(to)) and delayed outward K(+) current (I(K)) in isolated cardiac myocytes was determined using the whole cell patch clamp technique in guinea pig and dog. RP62719 decreased I(K1) with an inhibitory concentration 50 (IC(50) ) of 5.0+/-1.0 micromol/L at -100 mV in guinea pig ventricular cells. In dog ventricular myocytes, RP62719 inhibited Ito by 84+/-4.4% with an IC(50) of 1.2+/-0.51 micromol/L at +40 mV. In guinea pig ventricular cells, RP62719 decreased I(K): I(Kstep) by 50.0+/-8.3%%and I(Ktail) by 56.0+/-4.9% at +40 mV, respectively. RP62719 inhibited I(Kstep) with an IC(50) of 4.2+/-0.8 micromol/L and I(Ktail) with an IC(50) of 3.3+/-0.75 micromol/L. Thus it is suggested that the ionic mechanism of antiarrhymic effect by RP62719 may be due to its inhibition of I(K1),I(to) and I(K).

[Effects of 7-bromoethoxybenzene-tetrahydro-palmatine on isolated working guinea pig heart].

The effective period of isolated working guinea pig heart perfused with Tris-sodium-pyruvate solution (Tris-SP) was shown to be 70 min. Perfluorotributylamine at a concentration of 0.5% added to Tris-SP prolonged the effective time to 90 min. 7-Bromoethyoxybenzene-tetrahydropalmatine (EBP) 10 mumol/L, and prazosin 0.1 mumol/L showed no obvious influence on the parameters in the isolated working guinea pig heart. Methoxamine 1 mumol/L and dopamine 1 mumol/L produced marked changes on LVP, ABF, +/- dp/dtmax and T-CO. It is suggested that the alpha 1-adrenoceptor was partially associated with the positive chronotropic action of the heart. Effects of methoxamine and dopamine on the parameters were antagonized by EBP. It shows that the antagonistic affects of EBP on methoxamine and dopamine are concerned with its antagonistic effect on alpha 1-adrenoceptor and dopamine receptor. This is consistent with previous results in rabbit aortic strips and rat anococcygeus muscles.

[The mechanism of antiarrhythmic action of 7-bromoethoxybenzene-tetrahydropalmatine].

The mechanism of antiarrhythmic action and the electrophysiologic effects of 7-bromoethoxybenzene-tetrahydroptamine (EBP) have been studied using conventional microelectrode technique. The effect of EBP on the membrane ISi and I(X) currents were investigated in the canine cardiac purkinje fibres using the double microelectrode voltage clamp methods. EBP was shown to increase the duration of action potential at 20 and 90% of repolarization of isolated guinea pig papillary muscles. However, the amplitude of action potential, the resting potential and the overshoot and maximum rate of 0 phase depolarization (Vmax) remained unchanged. The membrane potential was held at 40 mV and command potential at -15 mV, 0.3 Hz 500 ms. After 10 min of perfusion of EBP 30 mumol/L, the slow inward current (ISi) was reduced markedly. When the holding potential was held at -20 mV, and the command potential at +10 mV, EBP (10-100 mumol/L) exerted a depressed effect on the delayed (outward) rectifier current (I(X)) in a dose-dependent manner. It may be concluded that the antiarrhythmic effects of EBP is mainly related to decrease the automaticity and prolong the duration of action potential at 20-90% of repolarization of isolated guinea pig papillary muscles.

[Effects of fluid percussion injury on intracellular [Ca2+]i and pH in cultured rat neurons].

AIM: To study the change of intracellular [Ca2+]i and pH in cultured neurons after fluid percussion injury, and the therapeutic effect of drugs. METHODS: The neurons of Sprague Dawley rats were cultured for 8-14 days, then treated them with fluid percussion injury (2.5 kPa, 20 ms). Alterations of [Ca2+]i and pH in single neural cells following fluid percussion injury were measured by a laser scanning confocal microscope. After being injured for several hours the cultured neurons were treated with nimodipine or D-(-)-2-amino-5-phosphonovaleric acid (D-AP-5). Two hours later, the effects of drugs on intracellular [Ca2+]i and pH were studied. RESULTS: The Intracellular [Ca2+]i increased quickly after brain injury and reached peak in 12 hours. It then decreased gradually and became normal at 48 hours. The pH decreased slowly, reached minimum in 12 hours, and then kept at a lower level. It did not recover normal at 48 hours. Nimodipine and D-AP-5 decreased significantly the ascension of [Ca2+]i and the descent of pH. But nimodipine and D-AP-5 must be given within 10 hours after injury for a good therapeutic effect. CONCLUSION: According to the change of intracellular [Ca2+]i and pH, early use of nimodipine and D-AP-5, will get a better therapeutic effect.

[Cardiac-hemodynamic effects of M3 receptor agonist on rat and rabbit hearts].

AIM: To study the activation of choline on M3-R in heart and observe the hemodynamic changes of rat and rabbit. METHODS: A cardiac catheter was inserted into the left ventricular cavity via the right carotid artery, then the HR, LVSP, LVEDP, and +/- dp/dt were measured using a polygraph system. RESULTS: Choline was shown to decrease the hemodynamic assessments, such as HR, + dp/dt, LVSP and LVEDP. while the M3-R antagonist 4-DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) showed little effect on these assessments. It was found to reverse the hemodynamic effects of choline. CONCLUSION: M3 receptor agonist can produce negative inotropic and chronotropic effects on the heart of rat and rabbit.

Ketamine-induced ventricular structural, sympathetic and electrophysiological remodelling: pathological consequences and protective effects of metoprolol.

BACKGROUND AND PURPOSE: Growing evidence suggests that long-term abuse of ketamine does harm the heart and increases the risk of sudden death. The present study was performed to explore the cardiotoxicity of ketamine and the protective effects of metoprolol. EXPERIMENTAL APPROACH: Rats and rabbits were divided into control, ketamine, metoprolol alone and ketamine plus metoprolol groups. Ketamine (40 mg·kg(-1) ·day(-1), i.p.) and metoprolol (20 mg·kg(-1) ·day(-1), p.o.) were administered continuously for 12 weeks in rats and 8 weeks in rabbits. Cardiac function, electrophysiological disturbances, cardiac collagen, cardiomyocte apoptosis and the remodelling-related proteins were evaluated. KEY RESULTS: Rabbits treated with ketamine showed decreased left ventricular ejection fraction, slowed ventricular conduction velocity and increased susceptibility to ventricular arrhythmia. Metoprolol prevented these pathophysiological alterations. In ketamine-treated rats, cardiac collagen volume fraction and apoptotic cell number were higher than those of control animals; these effects were prevented by co-administration of metoprolol. Consistently, the expressions of poly (ADP-ribose) polymerases-1, apoptosis-inducing factor and NF-κB-light-chain-enhancer of activated B cells were all increased after ketamine treatment and sharply reduced after metoprolol administration. Moreover, ketamine enhanced sympathetic sprouting, manifested as increased growth-associated protein 43 and tyrosine TH expression. These effects of ketamine were prevented by metoprolol. CONCLUSIONS AND IMPLICATIONS: Chronic treatment with ketamine caused significant ventricular myocardial apoptosis, fibrosis and sympathetic sprouting, which altered the electrophysiological properties of the heart and increased its susceptibility to malignant arrhythmia that may lead to sudden cardiac death. Metoprolol prevented the cardiotoxicity of ketamine, indicating a promising new therapeutic strategy.

Receptor tyrosine kinases as therapeutic targets in malignant glioma.

Malignant gliomas have retained their dismal prognosis despite aggressive multimodal conventional therapeutic approaches, illustrating the need for novel therapeutic strategies. Recent advances in the cellular and molecular biology of gliomas have enhanced our understanding of the role of receptor tyrosine kinases (RTK) and RTK-mediated signal transduction pathways in tumor initiation, maintenance, angiogenesis, and vascular proliferation. Special attention has been focused on targets such as epidermal growth factor receptors (EGFR), platelet-derived growth factor receptors (PDGFR), vascular endothelial growth factor receptors (VEGFR), and on pathways such as the Ras/Raf/mitogen-activated protein (MAP)-kinase and phosphatidylinositol-3 kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathways. Novel targeted drugs known as small molecule inhibitors have been shown to modify the activity of these receptors and signaling pathways. Thus far, however, small molecule RTK inhibitor development has concentrated on a few RTK only, and drug activity has been comprehensively evaluated only in a limited number of different malignancies. One of the limiting factors for novel drug design and development is the incomplete knowledge of RTK functions in malignant glioma. This review summarizes current basic and clinical knowledge on the role of RTK in malignant glioma and on their importance as targets for new forms of therapy.

Frequency- and voltage-dependent inhibition of delayed outward potassium current by flecainide in isolated atrial cell of guinea pig heart.

Effects of flecainide (Fle) on membrane currents were studied using an isolated single atrial cell from guinea pig hearts. The tight-seal cell clamp technique was used. In the current clamp condition, Fle prolonged significantly the atrial action potential (APD) with frequency dependence. Delayed outward K+ current and outward tail current were specifically inhibited by Fle in a frequency- and concentration-dependent fashion. Fle inhibited Ik more strongly as the membrane potential became more positive from +10 mV to +60 mV. The value of Ik was attenuated to 973 pA from 1105 pA of control and the value of tail current was reduced to 113 pA from 288 pA of control at 60 mV. The drug did not affect the holding current. The effects of Fle on the action potential and transmembrane ionic currents strongly suggested that the main mechanism of action of this agent was to inhibit the voltage-dependent potassium current. In the voltage clamp condition, Fle affected neither the conventional L type Ca2+ current nor the Ik1 current significantly. Our research proved that Fle was not completely consistent with the class Ic agents, because Fle could markedly increase the APD in the experiment.

Effects of berbamine on hemodynamics and myocardial reperfusion injury in isolated working rabbit hearts.

The isolated working rabbit hearts perfused with Tyrode's buffer solution persisted working in good condition at least up to 70 min. Berbamine (Ber) 3 mumol.L-1 significantly changed the hemodynamic parameters and reduced cardiac functions. The effects of Ber appeared in a dose-dependent manner. Reperfusion following 30-min of global ischemia aggravated the myocardial damages induced by ischemia, and the parameters of cardiac functions in working hearts markedly reduced and did not restore even at 20 min of reperfusion. Ber 1 mumol.L-1 reduced the myocardial ischemic reperfused damages and restored all parameters to the level of preischemia within 10 min of reperfusion, and this situation of working hearts also lasted 40-50 min. The present results showed that Ber could protect myocardium against ischemic reperfused damages, promote the recovery of cardiac functions and prolong the efficient working period in isolated working rabbit hearts.

Effects of RP58866 on transmembrane K+ currents in mammalian ventricular myocytes.

AIM: To determine effects of RP58866 on inward rectifier K+ current (IKl), transient outward K+ current (Ito) and delayed outward rectifier K+ current (IK) in isolated cardiac myocytes. METHODS: In isolated ventricular myocytes of guinea pig and dog, the effect of RP58866 on IKl, Ito, and IK were observed by the whole cell voltage-clamp technique. RESULTS: RP58866 decreased IKl in a concentration-dependent manner, with an IC50 of (3.4 +/- 0.8) micromol.L-1 (n = 6) at -100 mV in guinea pig ventricular cells. In dog ventricular myocytes, RP58866 inhibited Ito with IC50 of (2.3 +/- 0.5) micromol.L-1 at +40 mV. In guinea pig ventricular cells, RP58866 at 100 micromol.L-1 decreased IK: IKstep by (58 +/- 13)% at +40 mV, and IKtail by (86 +/- 17)%, respectively. RP58866 inhibited IKstep with an IC50 of (7.5 +/- 0.8) micromol.L-1, and IKtail with an IC50 of (3.5 +/- 0.9) micromol.L-1. The envelope of tail analysis suggested that both IKr and IKs were inhibited. CONCLUSION: RP58866 inhibits IKl, Ito, and IK in cardiac myocytes with a similar potency, and is not a specific IKl inhibitor.

Inhibitory effects of berberine on IK1, IK, and HERG channels of cardiac myocytes.

AIM: To study the effects of berberine on inward rectifier potassium current (IK1) and outward delayed rectifier potassium current (IK) of guinea pig ventricular myocytes, and on human ether-a-go-go related gene (HERG) channel expressed in Xenopus oocytes. METHODS: Whole cell patch-clamp and geneclamp techniques were used to record ionic currents. RESULTS: Berberine prolonged action potential duration (APD) and inhibited IK1 and IK in a concentration-dependent manner. Berberine 100 micromol/L increased APD90 from (450 +\- 48) ms to (888 +\- 90) ms (n = 6, P < 0.01), and inhibited IK1 by 65 % +\- 7 % (n = 6, P < 0.01). Berberine 50 micromol/L inhibited IK by 57 % +\- 6 %, IKtail by 53 % +\- 6 % (n = 6, P < 0.01). Berberine produced a voltage-dependent block on IK that increased with stronger depolarization, and once all channels were activated, there was no further block at positive potentials. Berberine blocked the HERG channels potently with an IC50 value of approximately 75 micromol/L. This block was voltage-dependent, suggesting that it probably bind to either open or inactivated HERG channels. CONCLUSION: Berberine prolonged APD and possessed blocking effect on IK1, IK, and HERG channel expressed in Xenopus oocytes. The antiarrhythmic mechanism of berberine is related to its inhibitory effects on IK1, IK, and HERG channel.

Artemisinin blocks activating and slowly activating K+ current in guinea pig ventricular myocytes.

AIM: To study the effect of artemisinin (Art) on outward rectifier potassium current in ventricular myocytes. METHODS: In isolated guinea pig ventricular myocytes, the effects of Art on the two components of delayed outward rectifier K+ current (IK), the rapidly activating inward K+ current (IKr), and the slowly rectifying outward K+ current (IKs) were observed by the whole cell patch-clamp technique. RESULTS: Art decreased IK in a concentration-dependent manner. The IKstep and IKtail were reduced from 387 +/- 46 pA to 240 +/- 48 pA and from 299 +/- 30 pA to 130 +/- 38 pA, respectively at holding potential of +40 mV by Art 50 mumol.L-1. The envelope of tail analysis suggested that both IKr and IKs were inhibited. CONCLUSION: Art blocked the two components of delayed outward rectifier K+ current (IKr and IKs) in guinea pig ventricular cells.