Volume 121  ·   Issue 04  ·  2008

The incidence of prostate cancer(PCa)in Western developed countries is higher than in Asia where it is presently increasing rapidly.1 Data from Shanghai and Macau2 suggest that the incidence of PCa in the Chinese population has also shown an increasing trend over the last twenty years。

Abstract：Background Recent studies have shown that effective ingredients of Chinese herbs are used more and more widely in the treatment or co-treatment of cancers,however,they are usually used separately and there has been limited research about joint application of Chinese herbs in multi-modal treatment.The aim of this study was to screen a QHF(Q:Qingrejiedu,H:Huoxuehuayu and F:Fuzhengguben)formula for effective ingredients from Chinese medicines and assess its anti-hepatic cell cancer(HCC)effect in combination with chemotherapy.Methods Six effective ingredients from Chinese medicine were selected based on the previous literature and used in the study.The QHF formula and the best ratio of ingredients were evaluated in H22 mouse(KM)models with solid tumors and ascites tumors by uniform design and monitoring inhibition of tumor growth and survival.We then observed the anti-hepatic cell cancer(HCC)effect of QHF when combined with cisplatin(DDP)in H22 mouse(Balb/c)models with solid tumors and ascites tumors.Evaluating of the therapeutic effect included the general condition of the mice,inhibition of tumor growth,survival,changes in body weight,thymus index,spleen index and WBC counts.Results The optimal QHF dose ratio for anti-hepatic cell cancer treatment was:800 mg/kg Cinobufotalin,14 mg/kg Ginsenosides Rg3,5.5 mg/kg PNS and 100 mg/kg Lentinan.Treatment was more efficient in inhibiting the growth of transplanted tumors in H22 mice when using the QHF formula(55.91%)than using Cinobufotalin(33.25%),Ginsenosides Rg3(35.11%),PNS(27.12%)or Lentinan(4.97%)separately.QHF also prolonged the life of H22 ascites hepatic cancer mice more efficiently(38.13%)than Cinobufotalin(25.00%),Ginsenosides Rg3(27.27%),PNS(23.30%) or Lentinan (24.43%).QHF combined with DDP could reduce DDP-induced leucopenia,spleen and thymus atrophy and other toxic reactions.Combining QHF with DDP the tumor growth inhibition reached 82.54% with a 66.83% increase in survival.Conclusions QHF is more efficient in anti-hepatic cell caner treatment than the single drugs that constitute the formula.QHF combined with DDP can attenuate tumor growth and suppresses the DDP-induced toxic reactions。

Abstract：Background β2-adrenoceptor(β2AR)desensitization is a common problem in clinical practice.β2AR desensitization proceeds by at least such three mechanisms as heterologous desensitization,homologous desensitization and a kind of agonist-induced rapid phosphorylation by a variety of serine/threonine kinases.It is not clear whether there are other mechanisms.This study aimed to investigate potential mechanisms ofβ2AR desensitization.Methods Twenty-four BALB/c(6-8 weeks old)mice were divided into three groups,which is,group A,phosphate buffered saline(PBS)-treated;group B,ovalbumin(OVA)-induced;and group C,salbutamol-treated.Inflammatory cell counts,cytokine concentrations of bronchoalveolar lavage fluid(BALF),pathological sections,total serum IgE,airway responsiveness,membrane receptor numbers and total amount of β2AR were observed.Asthmatic mouse model and β2AR desensitization asthmatic mouse model were established.Groups B and C were selected for two-dimensional gel eleclrophoresis(2DE)analysis so as to find key protein spots related to β2AR desensitization.Results Asthmatic mouse model and β2AR desensitization asthmatic mouse model were verified by inflammatory cell count,cytokine concentration of BALF,serum IgE level,airway hyperreactivity measurement,radioligand receptor binding assay,Western blot analysis,and pathologic examination.Then the two groups(groups B and C)were subjected to 2DE.Two key protein spots associated with β2AR desensitization,Rho GDP-dissociation inhibitor 2(RhoGDI2)and peroxiredoxin 5,were found by comparative proteomics(2DE and mass spectrum analysis).Conclusion Oxidative stress and small G protein regulators may play an important role in the process of β2AR desensitization。

Abstract：Background Cell transplantation for myocardial repair is limited by early cell death.Gene therapy with human growth hormone(hGH)has been shown to promote angiogensis and attenuate apoptosis in the experimental animal.This study was conducted to explore the effects of myoblast-based hGH gene therapy on heart function restoration and angiogenesis after myocardial infarction,and to compare the differences between myoblast-based hGH gene therapy and myoblast therapy.Methods Myoblasts were isolated from several SD rats,cultured,purified,and transfected with plasmid pLghGHSN and pLgGFPSN.Radioimmunoassay(RIA)was used to detect the expression of hGH in these myoblasts.SD rats underwent the ligation of the left anterior descending coronary artery so as to establish a heart ischemia model.Thirty surviving rats that underwent ligation were randomly divided into 3 equal groups 2 weeks after left coronary artery occlusion:pLghGHSN group received myoblast infected with hGH gene transplantation;pLgGFPSN group received myoblast infected with GFP gene transplantation;control group:received cultured medium only.Four weeks after the injection the surviving rat underwent evaluation of cardiac function by echocardiography.The rats were killed and ventricular samples were undergone immunohistochemistry with hematoxylin-eosin and factorⅧ.Cryosection was analyzed by fluorescence microscopy to examine the expression of green fluorescent protein.Reverse transcriptase-polymerase chain reaction(RT-PCR)was used to examine the mRNA expression of vascular endothelial growth factor(VEGF),bax and Bcl-2.hGH expression in myocardium was examined by Western blot.Results Myoblast can be successfully isolated,cultured and transfected.The expression of hGH in transfected myoblast was demonstrated with RIA.Four weeks after therapy,the cardiac function was improved significantly in pLghGHSN group and pLgGFPSN group.Fractional shortening(FS)and ejection fraction(EF)in pLghGHSN group were elevated significantly compared with pLgGFPSN group and control group after therapy(FS:36.9±5.3 vs 29.5±3.5,21.8±2.9;EF:56.9±4.3 vs 47.1±3.6,38.4±4.8,P＜0.05).Left ventricular end-diastolic dimension(LVEDD)and heart infracted size in pLghGHSN group were decreased significantly compared with pLgGFPSN group and control group after therapy(LVEDD:5.9±0.3 vs 6.8±0.2,8.6±0.3;heart infracted size:(34.5±4.2)%vs(40.0±3.9)%,(46.1±3.8)%,P＜0.05);Green fluorescence was detected in cryosection of pLgGFPSN group.The capillary density of the pLgGFPSN group was significantly greater than those of the pLghGHSN group and control group(P＜0.05).The mRNA expression of VEGF and Bcl.2/bax in pLghGHSN group was higher than in pLgGFPSN group or control group(P＜0.05).The expression of hGH gene in myocardium tissue can be detected by Western blot assay in pLghGHSN group.Conclusions Transplantation of heart cells transfected with hGH induced greater angiogenesis and effect of antiapoptosis than transplantation of cells transfected with GFP.Combined GH gene transfer and cell transplantation provided an effective strategy for improving postinfarction ventricular function。

Abstract：Background Isoflurane,a commonly used inhaled anesthetic,induces apoptosis in primary rat cortical neurons of rat in a concentration-and time-dependent manner by an unknown mechanism.We hypothesized that isoflurane induced apoptosis by causing abnormal calcium release from the endoplasmic reticulum(ER)via activation of inositol 1,4,5-trisphosphate(IP3)receptors.Sevoflurane has a reduced ability to disrupt intracellular calcium homeostasis and is aless potent cytotoxic agent.This study examined and compared the cytotoxic effects of isoflurane and sevoflurane on rat primary cortical neurons and their relationship with disruption of intracellular calcium homeostasis and production of reactive oxygen species(ROS).Methods Primary rat cortical neurons were treated with the equivalent of 1 minimal alveolar concentration(MAC)of isoflurane and sevoflurane for 12 hours.MTT reduction and LDH release assays were performed to evaluate cell viability.Changes of calcium concentration in the cytosolic space,[Ca2+]c,and production of ROS were determined after exposing primary rat cortical neurons to isoflurane and sevoflurane.We also determined the effects of IP3 receptor antagonist xestospongin C on isoflurane-induced cytotoxicity and calcium release from the ER in primary rat cortical neurons.Results Isoflurane at 1 MAC for 12 hours induced cytotoxicity in primary rat cortical neurons,which was also associated with a high and fast elevation of peak[Ca2+]c.Xestospongin C significantly ameliorated isoflurane cytotoxicity in primary cortical neurons,as well as inhibited the calcium release from the ER in primary cortical neurons.Isoflurane did not induce significant changes of ROS production in primary rat cortical neurons.Sevoflurane,at equivalent exposure to isoflurane,did not induce similar cytotoxicity or elevation of peak[Ca2+]c in primary rat cortical neurons.Conclusion These results suggested that isoflurane induced elevation in[Ca2+]c,partially via elevated activity of IP3 receptors,which rendered cells vulnerable to isoflurane neurotoxicity.ROS production was not involved in isoflurane-induced neurotoxicity.Sevoflurane,at an equivalent exposure to isoflurane,did not induce similar elevations of [Ca2+]c or neurotoxicity in primary cortical neu rons of rat。

Abstract：Background In a suitable microenvironment,bone marrow mesenchymal stem cells(BMSCs)can transdifferentiate into myocardial cells whose special gene can be expressed as structural proteins.Growth factor(GF)plays an important role in the cell migration,survival and differentiation.However,the effect of GF on the cellular differentiation is not well understood.In this study,the hepatocyte growth factor(HGF)and insulin like growth factor-1(IGF-1)were used in the mixed culture of BMSCs and myocardial cells and the effects of these growth factors on the GATA-4 expression of BMSCs were investigated.Methods BMSCs were isolated from the marrow of rabbit femurs and tibias and foetal rabbit ventricular myocytes were isolated with trypsin sequential digestion.These two kinds of cells were cocultured in a ratio of 1:1 for 6 weeks;cocultured cells with added HGF and IGF-1 were the experimental group.The differentiated BMSCs were collected using the laser capture,microdissection system and their RNA isolated.Immunocytochemical staining,transmission electron microscopy and reverse transcription-polymerase chain reaction were used to evaluate the transformation of the stem cells into cardiomyocytes like cells.Results When cultured separately,BMSCs did not express a-actin and the stem cells had many nucleoli.However,when cocultured with cardiomyocytes,BMSCs expressed α-actin and the cardiac transcription factor GATA-4 and showed cardiomyocyte like ultrastructure.In comparison with the control group,the experimental group exhibited the enhanced expression level of GATA-4.The GATA-4 expression of BMSCs increased gradually following the addition of HGF and IGF-1,reached the maximal level after two weeks and decreased slightly thereafter.Conclusions BMSCs can transdifferentiate into cardiomyocytes like cells and express the cardiac transcription factor GATA-4 after being cocultured with myocardial cells.HGF and IGF-1 can stimulate transdifferentiation of BMSCs into cardiac phenotype and enhance the expression of GATA-4.These results indicate that growth factors have a great potential in clinical cellular therapy。

Abstract：Background Hyaluronan(HA)is most likely associated with tumor invasion and metastasis.Studies have shown that HA levels are often increased in serum of patients with various malignant tumors.The purpose of this study was to determine the levels of serum hyaluronan in patients with oral cancer and evaluate the value of serum HA in adjuvant diagnosis,staging and monitoring treatment response in these patients.Methods Eighty-four hospitalized patients with oral cancer,65 patients with benign tumors in the oral and maxillofacial region and 67 healthy individuals were included in this investigation.Venous blood was collected from these patients and the healthy individuals before therapy.One week after therapy,venous blood was collected once again in 43 patients with oral cancer.Serum samples were obtained and serum HA levels examined.Results The serum HA concentration was significantly higher in oral cancer patients than in patients with benign tumors and in healthy controls(P＜0.05).The serum HA level in patients with stages Ⅲ and Ⅳ disease was higher than in patients with stages Ⅰ and Ⅱ disease,but there was no significant difference in the HA level between stages Ⅰ and Ⅱ nor between stages Ⅲ and Ⅳ(P＞0.05).After a complete treatment the HA levels in patients with oral cancer became lower than before treatment,but the difference was not significant(P＞0.05).Conclusions The results of this study suggest that the determination of HA levels may provide additional information in diagnosis of oral cancer,but its usefulness as an adjunct in clinical staging and in monitoring treatment response was limited。

Abstract：Background Evidence showed that both myocardium and blood vessels were damaged in dilated cardiomyopathy (DCM).However,the changes in arterial compliance,serum cytokines and circulating endothelial progenitor ceIls(EPC),and their correlations remain unknown.Methods Sixty-five DCM patients and 49 healthy volunteers were studied.Both large artery compliance(C1)and small artery compliance(C2)were measured with the CVProfilor DO-2020.Quantitative enzyme-linked immunosorbent assays (ELISAs)were used to measure the levels of vascular endothelial growth factor-A(VEGF-A)and VEGF receptor 2(VEGF-R2).Circulating EPC was assessed by EPC colony-forming assays and flow cytometry(CD133+/CD34+cells).Phagocytized Dil-acLDL and binded FITC-UEA-I were used to analyze endothelial lineage marker expression by immunofluorescence.Results Although C2 was markedly lower in DCM patients than in control group((3.8±1.8)ml/mmHg×100 vs(5.0±2.2)ml/mmHg×100,P＜0.0001),there was no statistically significant difference in C1 between the two groups(P＞0.05).Levels of VEGF-A,the numbers of colony-forming units(CFU)and the fractions of EPC were obviously highein DCM patients than in control group((127.61±39.5)pg/ml vs(58.8±42.9)pg/ml,P＜0.0001;(2.5±1.5)%vs(0.5±0.3)%,P＜0.05;23.5±12.8 vs 10.8±7.4,P＜0.01,respectively)and however,there was no significant difference in VEGF-R2 between two groups(P＞0.05).LgVEGF-A was positively correlated with the number of EPC-CFU(r=0.435;P＜0.05)and inversely correlated with C2(r=0.543;P＜0.001)in DCM patients.Conclusions The reduction of C2,a sensitive marker reflecting endothelial dysfunction,was observed in DCM patients and closely related to the increase in serum VEGF-A。

Abstract：Background BAFF,the B cell activation factor,is a member of the tumor necrosis factor(TNF)ligand family that binds to BCMA,TACI,and BAFF-R.Previous studies have shown that members of the TNF family are detected in human placental trophoblast cells,but the expression patterns of BAFF involved in human decidua and the differential expression of BAFF between normal pregnancy and miscarriage are still incompletely documented or unknown.This study was designed to investigate the expression of BAFF and BAFF-R in the trophoblast and decidua of normal early pregnant women and recurrent spontaneous abortion(RSA)patients.Methods Forty-five patients with RSA and 45 normal pregnant women were included in this study.By reverse transcriptase-polymerase chain reaction(RT-PCR),Western blotting and immunohistochemical experiments,we explored the expression of BAFF and BAFF-R in the maternal-fetal interface of normal early pregnant women and RSA patients.Results Analysis by RT-PCR and Western blotting revealed that BAFF was detected in both trophoblast and decidua of all the samples,and the expression level was higher in the tissues of normal early pregnant women(P＜0.05)than that of recurrent spontaneous abortion patients under the same gestational weeks.Messages for BAFF-R were absent.Immunohistochemical experiments showed that expression of BAFF was cell-specific which was localized to villous cytotrophoblast and syncytiotrophoblast cells in trophoblast and to stromal cells in decidua.Whereas BAFF was prominent on the trophoblast and decidua of normal early pregnant women,it was decreased in the tissues of RSA patients.Conclusions BAFF might steer maternal leukocytes away from a harmful immune response and toward a favorable one and play a potentially vital role for successful pregnancy。

Abstract：Background Variation in prostate cancer incidence between different racial groups has been well documented,for which genetic polymorphisms are hypothesized to be an explanation.We evaluated the association between polymorphisms in the cytochrome P-450 CYP1A1(CYP1A1)and glutathione S-transferase M1(GSTM1)genes and genetic susceptibility to prostate cancer in Chinese men.Methods Two hundred and eight prostate cancer patients and 230 age matched controls were enrolled in this study.All DNA samples from peripheral blood lymphocytes were genotyped for common genetic polymorphisms of the CYP1A1 and GSTM1 genes using the oligonucleotide microarray(DNA chip)technique and the polymorphism results confirmed by sequencing.The different polymorphisms in prostate cancer patients were also analyzed according to age at diagnosis,prostate specific antigen level,cancer stage and grade(Gleason score).Results The prevalence of the GSTM1(0/0)genotype was significantly higher in prostate cancer patients(58.2%)than in controls(41.7%,P＜0.05).Further analysis demonstrated that the prostate cancer patients with a GSTM1(0/0)genotype were younger than those with the GSTM1(+/+)genotype(P=0.024).No significant differences in the frequency distributions of CYP1A1 polymorphisms were observed between prostate cancer patients and controls.Conclusion GSTM1(0/0)gene polymorphism may be linked to prostate cancer risk and early age of onset in Chinese。

Abstract：Background Nucleostemin is essential for the proliferation and survival of stem and cancer cells,but it is unknown whether this newly identified molecule is involved in prostate cancer pathogenesis.Methods Total RNA and protein were extracted from prostate cancer tissues and PC-3,LNCap and DU145 cell lines.The nucleostemin mRNA and protein expression were measured by RT-PCR and Western blot.Immunohistochemistry was also used to detect the nucleostemin protein expression in prostate cancer tissues and PC-3 cells.A nucleostemin specific,short hairpin RNA,expression plasmid was used to transfect PC-3 cells.The changes of nucleostemin gene were detected and the proliferative capacity of the cells was determined.Results Nucleostemin was highly expressed in prostate cancer tissues and cell lines.Nucleostemin expression level in the silencer group PC-3 cells remarkably reduced.The proliferation rate of silencer group PC-3 cells decreased and the percentage of G1 stage cells increased.The neoplasm forming capacity in nude mice of the silencer group PC-3 cells decreased significantly.Conclusions Nucleostemin is highly expressed in prostate cancer tissues and cell lines.The proliferative capacity of PC-3 cells is remarkably reduced after silencing nucleostemin gene expression。

Abstract：Background All-trans retinoic acid(ATRA)can influence the tumor cell proliferation cycle,and some chemotherapeutic drugs are cycle specific.In this study,we hypothesize that ATRA can enhance chemotherapeutic drug sensitivity by affecting the cell cycle of tumor cells.Methods The cell cycle of LoVo cells was evaluated using flow cytometry(FCM).Cell viability was analyzed using the MTT assay.The morphologic changes in the treated LoVo cells were measured with acridine orange (AO)/ethidium bromide(EB)staining.Expression of survivin in LoVo cells was analyzed by immunofluorescence assay.Results After LoVo cells were treated with ATRA,the G0/G1 ratio of the tumor cells increased and the cell ratio of Sand G2/M-phase decreased.Viability of the cells decreased significantly after combined treatment with ATRA and 5-fluorouracil(5-FU)or mitomycin c(MMC) and was evaluated by fluorescence microscopy.Expression level of survivin in the tumor cells decreased after ATRA combination treatment.Conclusions ATRA enhances drug sensitivity of the LoVo cell line to cell cycle-specific agents and inhibits the expression of survivin in LoVo cells.The combination of ATRA and 5-FU or MMC promoted cell apoptosis,and the mechanism involved in apoptosis may be related to inhibition of survivin gene expression。

Abstract：Background The pelvis often needs to be reconstructed after bone tumor resection.A major challenge here for the orthopedic surgeons is to choose a method that gives the best performance which depends upon its biomechanical properties.In this study,a 3-dimensional finite element analysis(FEA)was used to analyze the biomechanical properties of reconstructed pelvis using fibula transplant fixed by four commonly used rod-screw systems.Methods A total pelvic finite-element model including the lumbar-sacral spine and proximal femur was constructed based on the geometry of CT image from a healthy volunteer.Three-dimensional finite element models of different implants including fibula,rod and screw were simulated using ways of solid modeling.Then various reconstructed finite element models were assembled with different finite element implant model and type Ⅰ resected pelvic finite element model.The load of 500 N was imposed vertically onto the superior surface of L3 vertebral body,and the pelvis was fixed in bilateral leg standing positions.FEA was performed to account for the stress distribution on the bones and implants.The pelvis displacement of the different rod-screw fixation methods and the maximum equivalent stress(max EQV)on all nodes and element were figured out to evaluate the advantages and disadvantages of different reconstructive methods.Results Stress concentration in the fibula transplant was extremely high in the reconstructed pelvis,but could be substantially decreased by internal fixation,which partially transferred the stress from the fibula to the rod-screw systems.High stress concentration was also found in the implants,especially in the connection sites between screw and rod.Among the four methods of fixation,a double rod system with L5-S1 pedicle and ilium screws(L5-S1 HR)produced the best performance:least stress concentrations and least total displacement.Conclusion According to the stability and stress concentration,the method of L5-S1 HR fixation combined with fibula transplantation is better than other fixation methods in pelvic reconstruction after type Ⅰ resection。

With the extensive utilization of PSA,digital rectum examination and transrectal ultrasound for screening in the aging population,the diagnosis of prostate cancer in China has markedly increased during the past years,particularly in developed regions。

Abstract：Objective With increased incidence of prostate cancer and an increased number of patients undergoing radical prostatectomy in China,it will be necessary to elaborate the diagnosis,clinical significance and treatment of patients whose tumors have positive surgical margins following radical prostatectomy.Data sources Positive surgical margin,prostate cancer and radical prostatectomy were used as subject words and the medical literature in recent decades was searched using the PubMed database and the results are summarized.Study selection Using positive surgical margin,prostate cancer and radical prostatectomy as subject words the PubMed medical database produced 275 papers of pertinent literature.By further screening 28 papers were selected and they represent relatively large-scale clinical randomized and controlled clinical trials.Results A pertinent literature of 275 papers was identified and 28 papers on large clinical studies were obtained.Analysis of results indicated that the positive rate of surgical margin after radical prostatectomy is 20%-40%,and although most patients with positive surgical margins are stable for a considerable period,the data available now suggested that the presence of a positive surgical margin will have an impact on the patient's prognosis.The risk factors of positive surgical margin include preoperative prostate specific antigen level,Gleason's score and pelvic lymph node metastasis.The most common site with positive surgical margin is in apical areas of the prostate;therefore surgical technique is also a factor resulting in positive surgical margins.From data available now it appears that as long as the surgical technique is skilled,different surgical modes do not affect the rate of surgical margin.Adjuvant radiotherapy is mainly used to treat patients with positive surgical margin after radical prostatectomy,but combination with androgen deprivation therapy may increase the curative effect.Conclusion The current data indicated that the presence of positive surgical margins can markedly affect the patient's prognosis.Therefore we should be aware how we reduce the positive surgical margin,how to diagnose positive surgical margin and how to treat when there are positive surgical margins。

Nowadays,with laparoscope techniques increasingly developed its indications are covering more complicated medical fields.Moreover,as a camera holder,the robotic system of an automated endoscopic system for optimal positioning (AESOP) can be controlled directly by the surgeon's voice。

Splenogonadal fusion is a rare congenital anomaly,which is described as abnormal fusion between the spleen and gonad or derivatives of the mesonephros.Surgical exploration is generally performed to rule out malignancy.The appearance at exploration can be misleading and often results in unnecessary orchiectomy。