Determination of total phenol and six polyphenolic components in the polyphenol extract of Cinnamomi cortex by quantitative nuclear magnetic resonance spectroscopy.

A quantitative nuclear magnetic resonance spectroscopy (qNMR) method was established for determining the total phenol and six polyphenolic components in the polyphenol extract of Cinnamomi cortex. The qNMR approach utilized DMSO-d6 as the deuterated solvent and potassium hydrogen phthalate as the internal standard for quantifying the total phenolic content, expressed as epicatechin equivalence in the sample. Two complementary qNMR methods with DMSO-d6 or D2O as solvent were established to simultaneously determine 6 polyphenol components in the cinnamon polyphenol extract, including epigallocatechin gallate (EGCG), epicatechingallate (ECG), epicatechin (EC), epigallocatechin (EGC), gallocatechin gallate (GCG) and gallic acid (GA). Method validation demonstrated excellent precision with intraday relative standard deviation (RSD) below 1.08% and interday RSD below 1.48%. The linear correlation coefficient (r) exceeded 0.999, and the limits of detection (LOD) were from 0.01 to 0.14 mg mL-1, while the limits of quantification (LOQ) were from 0.07 to 0.69 mg mL-1. Recovery rates for this method fell within the range of 98.2% to 101.7%. Furthermore, the method has been successfully applied for determining the polyphenolic content in authentic cinnamon polyphenol extracts obtained from different sources.

Acute liver failure: A systematic review and network meta-analysis of optimal type of stem cells in animal models.

BACKGROUND: The therapeutic effects of various stem cells in acute liver failure (ALF) have been demonstrated in preclinical studies. However, the specific type of stem cells with the highest therapeutic potential has not been determined. AIM: To validate the efficacy of stem cells in ALF model and to identify the most promising stem cells. METHODS: A search was conducted on the PubMed, Web of Science, Embase, Scopus, and Cochrane databases from inception to May 3, 2022, and updated on November 16, 2022 to identify relevant studies. Two independent reviewers performed the literature search, identification, screening, quality assessment, and data extraction. RESULTS: A total of 89 animal studies were included in the analysis. The results of traditional meta-analysis showed that stem cell therapy could significantly reduce the serum levels of alanine aminotransferase [weighted mean difference (WMD) = -181.05 (-191.71, -170.39)], aspartate aminotransferase [WMD = -309.04 (-328.45, -289.63)], tumor necrosis factor-alpha [WMD = -8.75 (-9.93, -7.56)], and interleukin-6 [WMD = -10.43 (-12.11, -8.76)] in animal models of ALF. Further subgroup analysis and network meta-analysis showed that although mesenchymal stem cells are the current research hotspot, the effect of liver stem cells (LSCs) on improving liver function is significantly better than that of the other five types of stem cells. In addition, the ranking results showed that the possibility of LSCs improving liver function ranked first. This fully proves the great therapeutic potential of LSCs, which needs to be paid more attention in the future. CONCLUSION: LSCs may have a higher therapeutic potential. Further high-quality animal experiments are needed to explore the most effective stem cells for ALF.

Sodium houttuyfonate protects against cardiac injury by regulating cardiac energy metabolism in diabetic rats.

Diabetic cardiomyopathy is a diabetic complication with complicated pathophysiological changes and pathogenesis and difficult treatment. Sodium houttuyfonate is the adduct of sodium bisulfite and houttuynin, the main volatile component in Houttuynia cordata Thunb, possesses a variety of activities including multiple interventions on inhibiting ventricular remodeling. The study aims to explore effect of sodium houttuyfonate on diabetic myocardial injury and its underlying mechanisms. The diabetes model was established by intraperitoneal injection of streptozotocin at a dose of 85 mg/kg. By intragastric administration for 26 days, sodium houttuyfonate (50 and 100 mg/kg/d) reversed the abnormal serum levels of triglyceride, total cholesterol, low-density lipoprotein cholesterol and low-density lipoprotein cholesterol to high-density lipoprotein cholesterol ratio, improved the abnormal levels of serum aspartate aminotransferase and brain natriuretic peptide, reduced electrocardiogram P-R and QRS interval extension, accelerated the heart rate, decreased serum malondialdehyde content, up-regulated the myocardial energy metabolism including elevated the contents of ATP, ADP, total adenine nucleotides and phosphocreatine in myocardium, decreased AMP/ATP ratio, elevated myocardial Ca2+-Mg2+-ATPase activity, and down-regulated the mRNA expressions of AMP protein activation kinase α2 (AMPK-α2) and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α). In a conclusion, these results suggest that sodium houttuyfonate can improve cardiac energy metabolism disorder caused by diabetes by increasing cardiac Ca2+-Mg2+-ATPase activity and regulating AMPK signaling pathway, and then attenuates cardiac injury caused by hyperglycemia. In addition, sodium houttuyfonate also has the effects of anti-oxidation and improving abnormal levels of blood lipid.

NFAM1 Promotes Pro-Inflammatory Cytokine Production in Mouse and Human Monocytes.

NFAT activating protein with ITAM motif 1 (NFAM1) is an ITAM bearing-transmembrane receptor that has been reported to play a role in B cell signaling and development. We performed expression analysis of NFAM1 using publicly available gene expression data sets and found that NFAM1 expression is significantly induced in intestinal biopsies from Crohn's disease (CD) and ulcerative colitis (UC) patients. At the cellular level, we further observed high expression of NFAM1 in monocytes and neutrophils, and low expression in B and T cells. To explore the role of NFAM1 in multiple immune cells and its potential role in IBD, we generated NFAM1-/- mice. In contrast with previous reports using NFAM1-transgenic mice, NFAM1-/- mice have no obvious defects in immune cell development, or B cell responses. Interestingly, NFAM1-/- monocytes produce reduced levels of TNF-α in response to activation by multiple IBD-relevant stimuli, including CD40L, TLR ligands and MDP. Additional cytokines and chemokines such as IL-6, IL-12, CCL3 and CCL4 are also reduced in CD40L stimulated NFAM1-/- monocytes. Collectively, these findings indicate that NFAM1 promotes monocyte activation, thereby amplifying the response to diverse stimuli. Similarly, we observed that deletion of NFAM1 in human monocytes reduces expression of CD40L-induced CCL4. Lastly, to assess the role of NFAM1 in IBD, we compared development of anti-CD40 induced colitis in NFAM1+/+ and NFAM1-/- mice. We found that although NFAM1 deletion had no impact on development of gut pathology, we did observe a decrease in serum TNF-α, confirming that NFAM1 promotes pro-inflammatory cytokine production in vivo. Taken together, we conclude that NFAM1 functions to amplify cytokine production and should be further evaluated as a therapeutic target for treatment of autoimmune disease.

Determination of adenosine phosphates in mouse myocardium tissue by HPLC with UV detection and using porous graphite carbon column.

A high-performance liquid chromatography (HPLC) method with UV detection was established and validated for the simultaneous determination of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) in mouse myocardial tissues. After protein precipitation and compound extraction with pre-cooled perchloric acid and the supernatant was centrifuged with the pH value adjusted to 6.5-7.5, the analytes were separated on a porous graphitic carbon LC column (4.6 mm × 100 mm, 5 µm) using gradient elution with a mobile phase of 10 mmol/L borax solution, pH 9.18(A) and acetonitrile-tetrahydrofuran (1:1, v/v) (B). The LC flow rate was 0.8 mL/min; the UV detection wavelength was 254 nm and the column temperature was maintained at 35 °C. ATP, ADP, and AMP were separated and the intra-day relative standard deviations (RSDs) of peak area repeatability were 1.3-2.5% (n = 6). The correlation coefficients of the linearity between UV responses and adenosine phosphate concentrations were larger than 0.9998 in all cases, within concentration ranges of 0.71-91.6 µg/mL for ATP, 1.3-81.5 µg/mL for ADP and 1.69-108.1 µg/mL for AMP. The limits of detection were within 0.17-0.21 µg/mL. The average standard substance spiked-in recoveries were 93.6-104.7% (n = 3). The established HPLC method was successfully applied to quantitate ATP, ADP, and AMP in mouse myocardial tissues.

Effect of fermented Cordyceps sinensis on doxorubicin­induced cardiotoxicity in rats.

Cordyceps sinensis (CS) is a prominent medicinal herb in traditional Chinese medicine, and fermented CS is frequently used as a substitute for natural CS. Doxorubicin (DOX), an antitumor drug used in chemotherapy, is limited by its poor cardiotoxicity. The aim of the present study was to evaluate the protective effect of fermented CS against DOX­induced cardiotoxicity and the potential underlying mechanisms. Male Sprague­Dawley rats (180­200 g) were randomly assigned to seven different treatment groups: Normal control, DOX control, DOX+captopril (0.05 g/kg), 0.75, 1.5 and 3 g/kg DOX+CS, and the CS (1.5 g/kg) control. Histopathological changes, cardiac energy metabolism, cyclic adenosine monophosphate (cAMP) signaling and the associated mRNA expression of AMP­activated protein kinase (AMPK) were then evaluated. Fermented CS decreased the left ventricular weight index, heart weight index and mortality; however, it increased diastolic blood pressure and mean arterial pressure. In addition, it shortened the duration of the QRS complex and Sα­T segment, decreased serum creatine kinase (CK) and aspartate aminotransferase activity, inhibited histopathological changes and reduced brain natriuretic peptide content. Treatment with fermented CS also increased the activities of superoxide dismutase and glutathione peroxidase, reduced malondialdehyde content, increased the mitochondrial activities of Na+K+­adenosine 5'­triphosphate (ATP) ase, Ca2+Mg2+­ATPase and CK, and increased the creatine phosphate/ATP ratio and AMP/ATP ratio. Furthermore, it decreased the ATP/adenosine 5'­diphosphate (ADP) ratio, upregulated AMPKα2 expression, reduced the activity of serum phosphodiesterases (PDEs) and increased myocardial cAMP content. The results of the present study demonstrated that fermented CS attenuated DOX­induced cardiotoxicity by inhibiting myocardial hypertrophy and myocardial damage, ameliorating systolic function and the antioxidant enzyme system, improving cardiac energy metabolism, depressing the activities of PDEs, and by upregulating the cAMP and AMPK signaling pathways. Thus, fermented CS may be a candidate for the prevention of DOX­induced cardiotoxicity, cardiac energy impairment and against a number of cardiac diseases.

Synergistic effects of polydatin and vitamin C in inhibiting cardiotoxicity induced by doxorubicin in rats.

The purpose of this study was to assess the synergistic effect of polydatin and vitamin C on attenuating cardiotoxicity induced by doxorubicin (DOX) in rats. Polydatin could significantly increase the activity of superoxide dismutase (SOD) and the heart rate, attenuate myocardial pathological damage, decrease malondialdehyde (MDA) content, slightly increase arterial pressure and glutathione peroxidase (GSH-Px) activity, reduce intervals of QRS, QT, and ST, and lower free fatty acid (FFA) content. The combination of polydatin and vitamin C could significantly increase arterial pressure and heart rate, decrease QRS interval and slightly reduce ST and QT intervals, significantly attenuate myocardial pathological damage, increase the activities of GSH-Px,T-SOD, Na+ K+ -ATPase, and Ca2+ Mg2+ -ATPase, elevate phosphocreatine (PCr) and adenosine triphosphate (ATP) contents, slightly increase adenosine diphosphate (ADP) and total adenine nucleotide (TAN) contents and PCr/ATP, and significantly decrease the contents of MDA and FFA, when compared with those in the DOX group. Meanwhile, the improvement effects on FFA content, the activities of ATPase and SOD, and contents of ATP and TAN in combination group were more obvious than those in polydatin group, and the improvement effects on arterial pressure, heart rate, interval of QRS, GSH-Px activity, and MDA, ADP, and PCr contents in combination group were slightly obvious when compared with those in polydatin group. In addition, the mRNA expression levels of AMPK-α2 and PPAR-α were slightly improved in combination group. The results illustrate that the combination of polydatin and vitamin C has the ability to enhance the myocardial protective effects by its antioxidative effect and improve energy metabolism.

Nanogold-functionalized g-C3N4 nanohybrids for sensitive impedimetric immunoassay of prostate-specific antigen using enzymatic biocatalytic precipitation.

This work reports on a new impedimetric immunosensing strategy for sensitive detection of prostate-specific antigen (PSA) in biological fluids. The assay was carried out on monoclonal anti-PSA capture antibody-modified glassy carbon electrode with a sandwich-type detection format. Gold nanoparticles-decorated g-C3N4 nanosheets (AuNP/g-C3N4), synthesized by the wet-chemistry method, were utilized for the labeling of polyclonal anti-PSA detection antibody and horseradish peroxidase (HRP). Upon target PSA introduction, the sandwiched immunocomplex could be formed between capture antibody and detection antibody. Followed by the AuNP/g-C3N4, the labeled HRP could catalyze 4-choloro-1-naphthol into benzo-4-chlorohexadienone. The as-generated insoluble product was coated on the electrode surface, thus increasing the Faradaic impedance of Fe(CN)6(4-/3)(-) indicator between the solution and the base electrode. Under the optimal conditions, the impedance increased with the increasing target PSA in the sample, and exhibited a wide linear range from 10pgmL(-1) and 30ngmL(-1) with a detection limit of 5.2pgmL(-1). A repeatability and intermediate precision of <14% was accomplished. The specificity and method accuracy in comparison with commercial PSA ELISA kit for analysis of human serum specimens were relatively satisfactory.

Doxorubicin toxicity changes myocardial energy metabolism in rats.

BACKGROUND: Doxorubicin (DOX) is an antitumor antibiotics used against malignancies. But its toxicity limits the therapy of DOX. OBJECTIVE: The purpose of this study was to evaluate DOX toxicity and the alteration of energy metabolism after short term and long term treatment. METHODS: Male Sprague-Dawley rats were randomly assigned to four groups: Short term control group, short term DOX treatment group, long term control group and long term DOX treatment group. In short term treated group, rats were injected with DOX i.p. at a dose of 2.5 mg/kg every 48 h for six equal injections. In long term, treated group, rats were tail-intravenously injected with DOX at a dose of 3 mg/kg once a week for four weeks. At the end of the experiment, histopathological changes, general blood biomarkers, endogenous antioxidant enzymes, cardiac energy metabolism and related mRNA expression of AMPK signal pathway were determined. RESULTS: DOX induced prominent oxidative stress, a higher mortality rate, histological and ECG changes, obvious cardiac hypertrophy, acute cardiac damage and cardiac energy impairment in short term treatment rats. In long term treatment rats, DOX caused serious nephropathy and systolic dysfunction, terrible cardiac energy impairment, clear alteration of substrate utilization and AMPK signal pathway. CONCLUSION: DOX treatment can induce different damages after short term and long term treatment. In short term treatment group, rats experienced a terrible mortality rate about 40%, the acute cardiac damage, cardiac energy impairment and an early heart failure which are potential connected with reduction of glucose utilization. In the long term treatment group, serious nephropathy and obvious changes of mRNA expressions of AMPK signal pathway were observed. Meanwhile, the serious cardiac energy impairment and substrate utilization alteration denote an obviously heart failure. This study could be helpful to develop therapy strategies of DOX complications for clinical application.

Comparative studies of polydatin and resveratrol on mutual transformation and antioxidative effect in vivo.

BACKGROUND: Polydatin and resveratrol are extractives of radix or rhizoma of Polygonum cuspidatum, and as the glycoside forms, it is a natural precursor of resveratrol. PURPOSE: In this study, we aimed to explore the mutual transformation between polydatin and resveratrol in rats, and to compare the antioxidative effect of them in vivo. STUDY DESIGN: In this study, we analyzed the serum molar concentration of polydatin and resveratrol after oral administration in rats and evaluated the anti-oxidative stress effects of them using a mouse model. METHODS: Rats were orally administered polydatin or resveratrol and the concentration of them in serum were analyzed by high performance liquid chromatography (HPLC). Their antioxidative effect was compared in mice with oxidative stress cardiomyopathy induced by doxorubicin (DOX). RESULTS: The results showed that polydatin and resveratrol could mutually transform in vivo, the molar concentration of polydatin in serum was always averagely 3.35 and 4.28 times as much as resveratrol after oral administration of polydatin and resveratrol at 200 mg/kg, respectively. Both polydatin and resveratrol could significantly decrease the content of malonydialdehyde (MDA), promote the activities of total superoxide dismutase (T-SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in plasma, and increase the content of glutathione (GSH) in myocardial tissue. The effect of polydatin surpassed resveratrol, particularly embodied in increasing the activities of T-SOD and CAT, and the content of GSH. CONCLUSION: It illustrates that polydatin is the main substance in serum after intragastric administration with polydatin or resveratrol, and the mutual transformation between polydatin and resveratrol keeps balance; they both have the ability of antioxidative stress in vivo, and polydatin has a better effect than resveratrol, which hints that polydatin may be a substitute for resveratrol in antioxidant for clinical use.

Transcriptome sequencing of Codonopsis pilosula and identification of candidate genes involved in polysaccharide biosynthesis.

BACKGROUND: Codonopsis pilosula (Franch.) Nannf. is one of the most widely used medicinal plants. Although chemical and pharmacological studies have shown that codonopsis polysaccharides (CPPs) are bioactive compounds and that their composition is variable, their biosynthetic pathways remain largely unknown. Next-generation sequencing is an efficient and high-throughput technique that allows the identification of candidate genes involved in secondary metabolism. PRINCIPAL FINDINGS: To identify the components involved in CPP biosynthesis, a transcriptome library, prepared using root and other tissues, was assembled with the help of Illumina sequencing. A total of 9.2 Gb of clean nucleotides was obtained comprising 91,175,044 clean reads, 102,125 contigs, and 45,511 unigenes. After aligning the sequences to the public protein databases, 76.1% of the unigenes were annotated. Among these annotated unigenes, 26,189 were assigned to Gene Ontology categories, 11,415 to Clusters of Orthologous Groups, and 18,848 to Kyoto Encyclopedia of Genes and Genomes pathways. Analysis of abundance of transcripts in the library showed that genes, including those encoding metallothionein, aquaporin, and cysteine protease that are related to stress responses, were in the top list. Among genes involved in the biosynthesis of CPP, those responsible for the synthesis of UDP-L-arabinose and UDP-xylose were highly expressed. SIGNIFICANCE: To our knowledge, this is the first study to provide a public transcriptome dataset prepared from C. pilosula and an outline of the biosynthetic pathway of polysaccharides in a medicinal plant. Identified candidate genes involved in CPP biosynthesis provide understanding of the biosynthesis and regulation of CPP at the molecular level.

[Isolation and mass spectrometric analysis of antioxidant peptides from enzymatic hydrolysates of roasted pills of Asini Corii Colla].

A fraction named GFC-1 with high antioxidant activities in vitro was isolated from the enzymatic hydrolysates of roasted pills of Asini Corii Colla, and the peptides in this fraction were identified. The enzymatic hydrolysates were isolated and purified with anion exchange chromatography and Sephadex G-25 filtration chromatography successively. GFC-1, a fraction isolated from the hydrolysates, exhibited the highest DPPH and ABTS scavenging capacity (DPPH 47. 95% at 2.0 g x L(-1) and ABTS 97.20% at 0.40 g x L(-1). Nine peptides from GFC-1 were identified by LC-ESI-MS/MS coupled with TurboSEQUEST search software and Swiss-Prot data base, and a high repetition core sequence GPAGPP*GPP* was also found.

[Calculus formation in the prostatic cavity after transurethral resection of the prostate: causes, treatment and prevention].

OBJECTIVE: To study the causes, clinical manifestations, treatment and prevention of calculus that develops in the prostatic cavity after transurethral resection of the prostate. METHODS: We reported 11 cases of calculus that developed in the prostatic cavity after transurethral resection or transurethral plasmakinetic resection of prostate. The patients complained of repeated symptoms of frequent micturition, urgent micturition and urodynia after operation, accompanied with urinary tract infection and some with urinary obstruction, which failed to respond to anti-infective therapies. Cystoscopy revealed calculi in the prostatic cavity, with eschar, sphacelus, uneven wound surface and small diverticula in some cases. After diagnosis, 1 case was treated by holmium laser lithotripsy and a second transurethral resection of the prostate, while the other 10 had the calculi removed under the cystoscope, followed by 1 -2 weeks of anti-infective therapy. RESULTS: After treatment, all the 11 cases showed normal results of routine urinalysis, and no more symptoms of frequent micturition, urgent micturition and urodynia. Three- to six-month follow-up found no bladder irritation symptoms and urinary tract infection. CONCLUSION: Repeated symptoms of frequent micturition, urgent micturition, urodynia and urinary tract infection after transurethral resection of the prostate should be considered as the indicators of calculus in the prostatic cavity, which can be confirmed by cystoscopy. It can be treated by lithotripsy or removal of the calculus under the cystoscope, or even a second transurethral resection of the prostate. For its prevention, excessive electric coagulation and uneven wound surface should be avoided and anti-infection treatment is needed.

[Downregulation of PAR expression induces the apoptosis of human prostate cancer PC3 cells and increases the Bcl-2/Bax ratio].

OBJECTIVE: To investigate the effects of the downregulated expression of the prostate androgen regulated (PAR) gene on the cell cycle and apoptosis of PC3 cells as well as on the expression level of Bcl-2/Bax. METHODS: After transfecting PC3 cells with small interfering RNA (siRNA) targeting PAR, we detected the inhibitory effect of PAR depletion on the proliferation of the PC3 cells by MTT assay, determined their apoptosis by flow cytometry, and measured the expression levels of Bcl-2 and Bax by Western blot. RESULTS: The expression of PAR was suppressed by siRNA, the G2-M phase PC3 cells were increased to (29.95 +/- 3.25)%, and the apoptosis of the cells was enhanced to (20.61 +/- 2.73)%, with statistically significant difference from the control group (P < 0.01). Western blot showed a decreased expression of Bcl-2, an increased expression of Bax, and an elevated ratio of Bax to Bcl-2. CONCLUSION: Downregulation of the PAR expression increases the Bax/Bcl-2 ratio and Bax expression, and thus induces the G2-M phase arrest and apoptosis of PC3 cells.

[Single nucleotide polymorphisms of CYP1A2 and their correlation with prostate cancer].

OBJECTIVE: To evaluate the correlation of the single nucleotide polymorphisms (SNPs) of the CYP1A2 gene with the stages and grades of prostate cancer (PCa). METHODS: We conducted gene sequencing of the rs2069514-3859 (A > G) and rs2069525-1707 (C >T) alleles in the CYP1A2 gene in 253 patients with benign prostatic hyperplasia (BPH) and 206 patients with PCa treated by castration therapy, and statistically analyzed their correlations with the genotypes, stages and grades of prostate cancer. RESULTS: The incidences of the 2 CYP1A2 SNPs showed no significant difference between the BPH and the castrated PCa patients (P > 0.05), and their genotypes were not correlated with the stages of PCa (P > 0.05). The Gleason scores were mostly <7 in the PCa patients with genotypes containing C in the rs2069525-1707 (C > T) allele (P = 0.030, OR = 4.658, 95% CI: 1.222 - 17.754). CONCLUSION: SNPs of the CYP1A2 gene may have some correlations with the pathologic stages of PCa, but their mechanisms and clinical significance need to be further confirmed.

Efficacy of a novel endotoxin adsorber polyvinylidene fluoride fiber immobilized with (L)-serine ligand on septic pigs.

A novel adsorber, polyvinylidene fluoride matrix immobilized with L-serine ligand (PVDF-Ser), was developed in the present study to evaluate its safety and therapeutic efficacy in septic pigs by extracorporeal hemoperfusion. Endotoxin adsorption efficiency (EAE) of the adsorber was firstly measured in vitro. The biocompatibility and hemodynamic changes during extracorporeal circulation were then evaluated. One half of 16 pigs receiving lipopolysaccharide (Escherichia coli O111:B4, 5 µg/kg) intravenously in 1 h were consecutively treated by hemoperfusion with the new adsorber for 2 h. The changes of circulating endotoxin and certain cytokines and respiratory function were analyzed. The 72 h-survival rate was assessed eventually. EAE reached 46.3% (100 EU/ml in 80 ml calf serum) after 2 h-circulation. No deleterious effect was observed within the process. The plasma endotoxin, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels were decreased during the hemoperfusion. Arterial oxygenation was also improved during and after the process. Furthermore, the survival time was significantly extended (>72 h vs. 47.5 h for median survival time). The novel product PVDF-Ser could adsorb endotoxin with high safety and efficacy. Early use of extracorporeal hemoperfusion with the new adsorber could reduce the levels of circulating endotoxin, IL-6, and TNF-α, besides improve respiratory function and consequent 72 h-survival rate of the septic pigs. Endotoxin removal strategy with blood purification using the new adsorber renders a potential promising future in sepsis therapy.

[Inhibitory effect of 5-aza-2'-deoxycytidine combined with docetaxel on prostate cancer PC3 cells in vitro].

OBJECTIVE: To evaluate the effects of methylation inhibitor 5-Aza-2'-Deoxycytidine (5-aza-2dc) and docetaxel (DT), alone or in combination, on the proliferation, migration, apoptosis and cell cycles of the human prostate cancer cell line PC3, and to investigate the possible mechanisms of these two drugs acting on prostate cancer in vitro. METHODS: Four groups were designed in this experiment: control, 5-aza-2dc, DT, and 5-aza-2dc + DT. The inhibitory effect of 5-aza-2dc and/or DT on the proliferation, migration and invasiveness of PC3 cells was detected by MTT, wound healing assay and cell migration assay, respectively. The apoptosis of the PC3 cells and its relationship with cell cycles were determined by Annexin V-FITC/PI assay and flow cytometry. RESULTS: 5-aza-2dc and/or DT significantly increased the inhibition rate of the PC3 cells, decreased their migration distance and reduced the number of the cells that invaded the lower chamber, most significantly in the 5-aza-2dc + DT group (P < 0.05). The cell apoptosis rates of the control, 5-aza-2dc, DT and 5-aza-2dc + DT groups were (10.65 +/- 0.39)%, (16.60 +/- 0.67)%, (17.95 +/- 1.08)% and (22.98 +/- 1.18)%, respectively, with the most significant increase in the combination group (P < 0.05). Combined medication of 5-aza-2dc and DT remarkably reduced the number of cells in the G0/G1 phase, and increased that in the G2/M phase (P < 0.05). CONCLUSION: 5-aza-2dc and DT, either alone or in combination, can significantly inhibit the proliferation, migration and invasiveness of PC3 cells in vitro, as well as induce their apoptosis and arrest their cell cycles in the G2/M phase, with even more significant effect when used in combination than applied alone.

[Effect of Chrysanthemum indicum on ventricular remodeling in rats].

OBJECTIVE: To explore the effects and mechanism of Chrysanthemum indicum on experimental ventricular remodeling induced by isoprenaline (ISO) and L-thyroxine (L-Thy). METHODS: The ventricular remodeling of mice were induced by subcutaneous injection of ISO with the dosage of 2 mg/kg daily for 7 d and the rats with L-Thy intraperitoneally with the dosage of 0.25 mg/kg daily for 9 d. After 7 days' treatment, the cardiac index and the Ang II content in myocardium of mice were measured. After 9 days' treatment, the ratios of LVW/BW, HW/BW of rats were calculated, the Ang II content in heart tissue and the ALD, TNF-alpha concentration in serun were determined by radioimmunoassay, the Hydroxy proline (Hyp) content in heart tissue were measured by hydrolysis method. RESULTS: After 7 - 9 days of treatment, Chrysanthemum indicum significantly reduced the left ventricular weight index and heart weight index in mice and rats with myocardial hypertrophy, decreased the content of Ang II in ventricular tissue in mice and rats, and reduced the ALD, TNF-alpha concentration in serum and the Hyp content in ventricular tissue in rats (P < 0.05). CONCLUSION: Chrysanthemum indicum can significantly attenuate the experimental ventricular remodeling; the mechanism may be related with restricting the activity of the sympathetic nervous system and decreasing the levels of Ang II, ALD and TNF-alpha.

[Penile and scrotal skin flaps: first choice for urethroplasty in the treatment of hypospadias].

OBJECTIVE: To explore the feasibility of the treatment of hypospadias with penile and scrotal skin flaps. METHODS: Twenty-three hypospadias patients aged 3.5-19 (mean 6. 8) years underwent urethroplasty with penile and scrotal skin flaps. All were followed up for 6 years and analyzed retrospectively. RESULTS: Of the total number of patients, 21 (91.3%) succeeded in one operation and the other 2 developed complications, including urethral fistula and urethral structure. CONCLUSION: Penile and scrotal skin, advantageous for its adequacy, rich blood supply and contribution to high success rate of surgery, is believed to be the first choice for urethroplasty in the treatment of hypospadias.

[Induction of prostate cancer-specific CTLs with dendritic cells pulsed by different types of tumor antigens].

OBJECTIVE: To study the effectiveness of freeze-thaw antigens and acid eluted peptide antigens extracted from tumor cell-pulsed dendritic cells (DC) in inducing prostate cancer-specific cytotoxic T lymphocytes (CTL) in vitro. METHODS: Tumor antigens were extracted from the prostate cancer cell line PC-3 with the repeated freeze-thaw and weak acid elution methods. Peripheral blood mononuclear cells were cultured with recombinant human GM-CSF and IL-4 for inducing DCs in vitro. Then the DCs were pulsed with the two kinds of prostate cancer tumor antigens respectively and cultured with T cells for inducing CTLs. The activity of the tumor-specific CTLs were detected by LDH release assay. RESULTS: The protein content in the tumor antigens obtained from PC-3 (2 x 10(7)) by citric acid-phosphate buffer elution and that by the repeated freeze-thaw method were (212.2 +/- 7.9) microg and (963.0 +/- 25.3) microg, respectively. The two kinds of prostate cancer antigens-pulsed DCs had a significant role in inducing the PC-3 cell-specific CTLs, and the CTLs induced by acid-eluted peptide antigen-pulsed DCs exhibited an even more significant tumor-specific cytotoxicity than those induced by repeated freeze-thaw ([60.4 +/- 5.52]% vs. [43.7 +/- 4.11]%, P < 0.01). CONCLUSION: Both the weak acid elution and repeated freeze-thaw methods for extracting prostate cancer antigens can be used for in vitro sensitization of DCs. The DCs pulsed by either of the two kinds of antigens can activate CTLs, and the antigens extracted by weak acid elution are even more effective.