Two novel chiral tetranucleate copper-based complexes: syntheses, crystal structures, inhibition of angiogenesis and the growth of human breast cancer in vitro and in vivo.

The single crystals of two novel chiral tetranucleate copper(II)-based complexes (TNCu-A and TNCu-B) containing L-methioninol-derived Schiff-bases were obtained. Their single structures were characterized by X-ray single crystal diffraction, infrared (IR) rays, elemental analysis, and liquid chromatography-mass spectrometry analysis. TNCu-A can effectively inhibit human umbilical vein endothelial cells (HUVECs) to form a tubular structure and it induces apoptosis of human triple-negative breast cancer MDA-MB-231 cells and HUVECs in vitro in a mitochondria dependent manner. Moreover, in vivo TNCu-A can remarkably inhibit the growth of triple-negative breast cancer from which MDA-MB-231 cells were xenografted into severely immunodeficient nude mice by inhibiting proliferation, inducing apoptosis of MDA-MB-231 cells by dramatically inhibiting the expression of the anti-apoptotic protein Bcl-2 and up-regulating the expressions of proapoptotic proteins caspase-9 and Bax, and simultaneously inhibiting tumor angiogenesis by decreasing the density of vascular endothelial cells and suppressing migration and even partially inducing apoptosis.

Early cardiopulmonary resuscitation on serum levels of myeloperoxidase, soluble ST2, and hypersensitive C-reactive protein in acute myocardial infarction patients.

BACKGROUND: Prompt and effective cardiopulmonary resuscitation (CPR) can promote the recovery of spontaneous circulation to some extent and can save patients' lives. The minimum target of cardiac resuscitation is the restoration of spontaneous circulation (ROSC). However, owing to prolonged sudden cardiac arrest, there is relatively high mortality within 24 h after cardiac resuscitation. Moreover, severe cerebral anoxia can deteriorate the prognosis of patients. Therefore, it is important to adopt an effective clinical evaluation of acute myocardial infarct (AMI) patients' prognosis after cardiac resuscitation for the purpose of prevention and management. AIM: To investigate early CPR effects on human myeloperoxidase (MPO), soluble ST2 (sST2), and hypersensitive C-reactive protein (hs-CRP) levels in AMI patients. METHODS: In total, 54 patients with cardiac arrest caused by AMI in our hospital were selected as the observation group, and 50 other patients with AMI were selected as the control group. The differences in serum levels of MPO, sST2, and hs-CRP between the observation group and the control group were tested, and the differences in the serum levels of MPO, sST2, and hs-CRP in ROSC and non-ROSC patients, and in patients who died and in those who survived, were analyzed. RESULTS: Serum levels of MPO, sST2, hs-CRP, lactic acid, creatine kinase isoenzyme (CK-MB), and cardiac troponin I (cTnI) were significantly higher in the observation group than in the control group (P < 0.05). Serum levels of MPO, sST2, hs-CRP, lactic acid, CK-MB, and cTnI in the observation group were lower after CPR than before CPR (P < 0.05). In the observation group, MPO, sST2, hs-CRP, lactic acid, CK-MB, and cTnI serum levels were lower in ROSC patients than in non-ROSC patients (P < 0.05). MPO, sST2, hs-CRP, and lactic acid serum levels of patients who died in the observation group were higher than those of patients who survived (P < 0.05). The areas under receiver operating characteristic curve predicted by MPO, sST2, hs-CRP, lactic acid, CK-MB, and cTnI were 0.616, 0.681, 0.705, 0.704, 0.702, and 0.656, respectively (P < 0.05). The areas under receiver operating characteristic curve for MPO, SST2, hs-CRP, and lactic acid to predict death were 0.724, 0.800, 0.689, and 0.691, respectively (P < 0.05). Logistic regression analysis showed that MPO, sST2, and hs-CRP were the influencing factors of ROSC [odds ratios = 1.667, 1.589, and 1.409, P < 0.05], while MPO, sST2, hs-CRP, and lactic acid were the influencing factors of death (odds ratios = 1.624, 1.525, 1.451, and 1.365, P < 0.05). CONCLUSION: Serum levels of MPO, sST2, hs-CRP, and lactic acid have a certain value in predicting recovery and prognosis of patients with ROSC.

Positive association between serum apolipoprotein M levels and hepatitis B virus DNA load in HBeAg-negative chronic hepatitis B.

BACKGROUND: Hepatitis virus B (HBV) has infected millions of people worldwide. Notably, such infections can be associated with hepatic complications. Levels of apolipoprotein M (apoM), a component of high-density lipoprotein (HDL), are known to be significantly elevated in patients with chronic hepatitis B (CHB). The aim of this study was to investigate the relationship between HBV DNA load in serum and serum apoM levels in patients with CHB. METHODS: A total of 73 HBeAg-negative CHB patients, 50 HBeAg-positive CHB patients, and 79 non-CHB controls were included in the study cohort. The age and body mass index (BMI) of the study participants were matched. Serum levels of apoM and the HBV antigens HBsAg and HBeAg were measured by enzyme-linked immunosorbent assay (ELISA) analysis. Serum levels of alanine aminotransferase (ALT), aspartate transaminase (AST), cholesterol, and triglycerides (TG) were assessed using an automatic biochemical analyzer. Serum HBV DNA levels were quantified by real-time PCR analysis. Data were analyzed by Spearman's rank correlation coefficient, Pearson correlation coefficient, and multivariate linear regression model (continuous variables), or Student's t-test (mean differences). RESULTS: Both the HBeAg-negative CHB and HBeAg-positive CHB patient groups exhibited elevated serum levels of apoM. Moreover, serum apoM levels were positively correlated with serum HBV DNA levels in HBeAg-negative CHB patients (r = 0.394, p < 0.001). Conversely, there was no significant relationship between apoM and HBV DNA levels in the HBeAg-positive CHB group (r = 0.197, p = 0.170). The median log copies/mL value for HBV DNA (4.00) was considered the cutoff point for the HBeAg-negative CHB group. Notably, a significant number of patients with HBV DNA levels above the cutoff point also had higher serum apoM levels (63.38 ± 29.84 vs. 41.41 ± 21.84; p = 0.001). CONCLUSIONS: Our findings reveal that the correlation between serum apoM levels and viral loads may depend on HBeAg status, as serum apoM levels were positively correlated with HBV DNA levels in HBeAg-negative CHB patients. These results suggest that HBeAg may play a role in apoM-related lipid metabolism and anti-inflammatory functions in hepatitis B patients. Thus, our findings may facilitate the clinical management of HBV infection.

Simultaneous determination of phenylalanine and tyrosine in peripheral capillary blood by HPLC with ultraviolet detection.

OBJECTIVES: Phenylalanine (Phe) and tyrosine (Tyr) are the most reliable indicators for the diagnosis of phenylketonuria (PKU). The purpose of this study is to establish a simple and rapid method for the determination of Phe and Tyr in peripheral capillary blood from newborns and children by high performance liquid chromatography with ultraviolet detection (HPLC-UV). METHODS: Peripheral blood specimens were taken from newborns or children by heel stick or skin puncture on the fingers. Blood samples were deproteinized by equal volume of 5% perchloric acid. Amino acid separation was carried out on a Hypersil C8 column. The mobile phase containing 5% acetonitrile (v/v) was run at a flow rate of 1.5 mL/min. Phe and Tyr were determined in the ultraviolet detector at 210 nm. RESULTS: The determination was within 10.0 min. The linear range was from 6.0 to 1512.0 µmol/L for Phe and 5.5 to 1250.0 µmol/L for Tyr. Although Phe and Tyr levels from peripheral blood samples were relatively lower than those from serum or plasma, the data showed a good agreement between them. CONCLUSION: The method we developed is simple, rapid and convenient. It is useful for screening and diagnosis of PKU in newborns and children.

Simultaneous determination of tyrosine, tryptophan and 5-hydroxytryptamine in serum of MDD patients by high performance liquid chromatography with fluorescence detection.

BACKGROUND: Tyrosine (Tyr), Tryptophan (Trp) and 5-hydroxytryptamine (5-HT) are important amino acids in vivo and have been hypothesized to be involved in many mental disorders. We developed a rapid and sensitive HPLC method for simultaneous measurement of serum Tyr, Trp and 5-HT and explored the clinical significances of Tyr, Trp and 5-HT and the 5-HT/Trp ratio for patients with major depressive disorder (MDD) disease. METHODS: Serum samples were deproteinized by 5% perchloric acid and separated on an Atlantis C18 column (4.6 × 150 mm, 5 µm) with the mobile phase consisting of 0.1 mol/l KH(2)PO(4) and methanol (85:15, V/V).The eluates were monitored by the fluorescence detection with programmed wavelength. RESULTS: Analysis was achieved in <12.0 min. The limits of quantification were 0.014, 0.005, and 0.024 µmol/l for Tyr, Trp and 5-HT, respectively. Reproducibility and recovery were satisfactory. Tyr, Trp and 5-HT and the 5-HT/Trp ratio were significantly decreased in patients with MDD. CONCLUSIONS: In diseases, like MDD, Tyr, Trp and 5-HT play an important role. This method can potentially be applied as prognostic or diagnostic tool or even to follow the evolution of the illness or of the treatment.

Clinical significance of simultaneous determination of serum tryptophan and tyrosine in patients with lung cancer.

BACKGROUND: To explore the clinical significance of serum tyrosine (Tyr) and tryptophan (Trp) in patients with lung cancer, we used a simple and efficient method of high-performance liquid chromatography with fluorescence detection (HPLC-FD) that simultaneously measured serum Trp and Tyr contents. METHODS: The concentrations of Tyr and Trp were measured simultaneously by HPLC-FD in the sera of 80 patients with lung cancer and 120 healthy controls. RESULTS: Trp concentrations were significantly lower in patients with lung cancer than in healthy controls (39.26±5.44 vs. 49.93±5.43 µmol/l, respectively; P<0.01), whereas in Tyr concentrations there were no differences with healthy controls (65.38±7.94 vs.66.40±8.55 µmol/l, respectively; P>0.05). In addition, patients in the adenocarcinoma group had significantly lower Trp and Tyr concentrations than those in squamous cell carcinoma group. There was no difference between the early stage and advanced stage of lung cancer. CONCLUSIONS: Determination of serum Trp and Tyr concentrations can be employed to assist the diagnosis of the histotypes of lung cancer and tumor stage. Tyr and Trp as indexes on the lung cancer diagnostic sensitivity, specificity were 54.9, 62.9% and 82.4, 92.1%, Trp is an important and special index for lung cancer diagnosis of which the specificity of diagnosis of lung cancer is more than 92%.

Simultaneous determination of serum tryptophan metabolites in patients with systemic lupus erythematosus by high performance liquid chromatography with fluorescence detection.

BACKGROUND: To provide a more comprehensive clinic marker of tryptophan (TRP) catabolism in patients with systemic lupus erythematosus (SLE), we developed a simple and efficient method that simultaneously measured serum TRP, kynurenine (KYN), and kynurenic acid (KYNA) using high performance liquid chromatography with fluorescence detection (HPLC-FD). METHODS: A simple and specific high performance liquid chromatography (HPLC) method was developed for simultaneously quantitative determination of TRP, KYN and KYNA with fluorescence detection (FD) using programmed wavelength and on-column fluorescence derivatization. Thirty patients with SLE and 80 healthy control subjects were analyzed for serum TRP metabolites using the assay we developed. The tryptophan breakdown index (TBI) and neuroprotective ratio (NPR) were calculated. RESULTS: The retention time of KYN, KYNA and TRP were 8.5 min, 13.7 min and 17.6 min, respectively. The linear range for TRP was 0.245-196 micromol/L, the limit of detection (LOD) was 0.001 micromol/L and average recovery was 103.71%. The linear range for KYN was 0.049-98 v/L, the LOD was 0.0245 micromol/L, and average recovery was 97.45%. The linear range for KYNA was 1.05-2093 nmol/L, the LOD was 0.05 nmol/L, and average recovery was 100.60%. Inter-day and intra-day relative standard deviations (SDs) were <5%. Phenylalanine, tyrosine, 5-hydroxytryptamine and creatinine did not interfere with the method. The results showed great differences in TRP, KYN and KYNA contents and TBI between patients with SLE and healthy controls, but little difference in NPR. CONCLUSIONS: The method is simple, fast, accurate, and meets the requirements for simultaneous determination of TRP, KYN and KYNA in serum.

[Expressions of eNOS and cytochrome P450 in the testis of sexually mature SD rats and their significance].

OBJECTIVE: To explore the expressions of endothelial nitric oxide synthase (eNOS) and cytochrome P450 (aromatase) in the testis of sexually mature male SD rats and their significance. METHODS: Eighteen male SD rats, 6 five-week, 6 seven-week and 6 ten-week old, were selected for this study. Paraffin sections of the left testis were made and the expressions of eNOS and P450 observed by the immunohistochemical ABC method. RESULTS: Positive expressions of eNOS and P450 were found to be + + +, + and + + in the Leydig cells of the five-week, seven-week and ten-week old rats, respectively, and they were also observed in a few spermatocytes, though with no regularity. CONCLUSION: In the Leydig cells of sexually mature male SD rats, eNOS and P450 are differently expressed in different stages of sexual maturation, and they are correlated as well.

[Expression and role of nitric oxide synthase in the testis and epididymis of Macaca fascicularis].

OBJECTIVE: To investigate the expression and the role of nitric oxide synthase (NOS) in the testis and epididymis of macaca fascicularis. METHODS: The immunohistochemical ABC method was used to observe the localization of nitric oxide synthase in the testis and epididymis of the macaca fascicularis. RESULTS: (1) nNOS immunoreactivity was found in the spermatogenic cells of seminiferous tubules, the epithelia of epididymal efferent ducts, sperm and the endothelia of blood vessels; (2) iNOS was expressed in the epididymal efferent duct, the sperm inside the duct, and the myoid cells and endothelia of blood vessels; (3) eNOS immunoreactivity was detected in the interstitial cells of the testis, the epididymal efferent duct, the sperm inside the duct, and the myoid cells and endothelia of blood vessels. CONCLUSION: NOS is extensively expressed in the testis and epididymis of the macaca fascicularis and it may play an important role in such processes as spermatogenesis, sperm maturation and testosterone secretion.

[Effects of glycosides of Tripterygium wilfordii, methyltestosterone and zhuanggushenjin capsule on nitric oxide synthase in rat testes].

OBJECTIVE: To investigate the effects of glycosides of tripterygium wilfordii (GTW), methyltestosterone and Zhuanggushenjin capsule on nitric oxide synthase (NOS) in rat testes. METHODS: Forty-five rats were equally divided into 5 groups, and respectively given GTW [10 mg/(kg x d)], methyltestosterone [2 mg/(kg x d)], Zhuanggushenjin capsule [0.3 g/(kg x d)], distilled water plus Tween 80 (control I), and distilled water alone (control II) for 4 weeks. At the end of the 5th week, the immunochemical ABC method was used to observe the effects of the three drugs on the NOS positive Leydig cells of the rats. RESULTS: Compared with control II, the GTW group had a significant decrease in the numbers of nNOS and eNOS positive Leydig cells, the methyltestosterone group showed an increase in the number of nNOS but a decrease in that of eNOS positive Leydig cells, and the Zhuanggushenjin group had an increase in the numbers of both nNOS and eNOS positive Leydig cells. CONCLUSION: GTW can reduce NO production by inhibiting eNOS and nNOS, and hence influence the spermatogenic process. Zhuanggushenjin capsule plays an important role in improving male sexual function by enhancing nNOS and eNOS expression and NO synthesis.