[Compound matrine injection reduces morphine tolerance of the mice with lung cancer by inhibiting expression of multidrug resistance gene 1 and P-glycoprotein].

Objective: To investigate the effect of compound matrine injection on morphine tolerance in mice with lung cancer in situ and the expressions of multidrug resistance gene 1 (MDR1) and P-glycoprotein (P-gp). Methods: A mouse model of lung cancer in situ and morphine tolerance mode was established. The mice were injected with gradient concentration of compound matrine. The pain thresholds under different conditions were measured by thermal radiation tail-flick method. The mRNA level of MDR1 was tested by reverse transcription polymerase chain reaction (RT-PCR) and the protein level of P-gp was detected by western blot. The DNA binding activity of cyclophosphoadenosine response element binding protein (CREB) to the promoter of MDR1 gene was detected by electrophoretic mobility shift assay (EMSA). Results: The maximum analgesic percentage (MPE) of the mice in the morphine group was (85.21±6.53)% on the 8th day, and decreased to (38.45±5.52)% and (28.14±4.52)% on the 10th and 12th day, respectively, which indicated the morphine tolerance of mice with lung cancer in situ.The MPE of the mice in the group treated with morphine and compound matrine injection (300 mg/kg) was (79.34±6.50)% on the 8th day, and decreased to (62.16±5.53)% and (40.20±4.50)% on the 10th and 12th day, respectively.The results of RT-PCR assay showed that the relative expression levels of MDR1 mRNA in the brain tissues of mice in the morphine group, saline group, morphine combined with compound matrine injection (300 mg/kg) group and compound matrine injection (200 mg/kg) group were 2.33±0.79, 1.04±0.38, 1.37±0.38, and 1.43±0.53, respectively. There were statistically significant differences between the morphine group and the normal saline group, the morphine group and the morphine combined with compound matrine injection (300 mg/kg) group (P<0.05). There was no significant difference between the normal saline group and the compound matrine injection (200 mg/kg) group (P=0.05). The results of western blot showed that the relative expression levels of P-gp protein in the brain tissue of mice in the morphine group, saline group, and morphine combined with compound matrine injection (300 mg/kg) group were 1.86±0.40, 1.00±0.23, and 1.27±0.27, respectively. The expression of P-gp protein in the morphine group was significantly higher than those of the normal saline group and the morphine combined with compound matrine injection (300 mg/kg) group (P<0.05). The DNA-binding activity of CREB in the saline group was (0.23±0.07) Pu, significantly lower than (0.89±0.23) Pu of morphine combined with naloxone group and (0.80±0.23) Pu of morphine group (P<0.05). While the CREB DNA binding activity of morphine combined with compound matrine injection (300 mg/kg) group was (0.79±0.21) Pu, implicated that compound matrine had marginal effect on the DNA-binding activity of CREB (P>0.05). Conclusion: Compound matrine injection can significantly improve morphine tolerance and drug resistance of lung cancer through inhibiting the upregulations of MDR1 and P-gp induced by morphine.

Pharmacokinetics of tulathromycin and its metabolite in swine administered with an intravenous bolus injection and a single gavage.

Tulathromycin is a macrolide antimicrobial agent proposed for therapeutic use in treatment of porcine and bovine respiratory disease. In this study, the absolute bioavailability of tulathromycin solution was investigated in pigs. Eight pigs, with body weight of 20.5 ± 1.6 kg, were given a single dose of tulathromycin at 2.5 mg/kg oral (p.o.) and intravenous (i.v.) in a crossover design. The plasma concentrations of tulathromycin and its metabolite were determined by LC-MS/MS method, and the pharmacokinetic parameters of tulathromycin were calculated by noncompartmental analysis. After p.o. administration, the maximum plasma concentration (C(max) ) was 0.20 ± 0.05 µg/mL at 3.75 ± 0.71 h. The terminal half-life (t(1/2λz) ) in plasma was 78.7 ± 6.75 h, and plasma clearance (Cl/F) was 1.14 ± 0.28 L/h/kg. After i.v. injection, plasma clearance (Cl) was 0.580 ± 0.170 L/h/kg, the volume of distribution (Vz) was 64.3 ± 21.2 L/kg, and the t(1/2λz) was 76.5 ± 13.4 h. In conclusion, an analytical method for the quantification of tulathromycin and its metabolite in plasma in swine was developed and validated. Following p.o. administration to pigs at 2.5 mg/kg b.w., tulathromycin was rapidly absorbed and the systemic bioavailability was 51.1 ± 10.2.

Histological analysis of regeneration of temporomandibular joint discs in rabbits by using a reconstituted collagen template.

The aim of this study was to design a biodegradable implant, in the form of a reconstituted collagen template in order to promote and support regeneration of the temporomandibular joint disc. Bovine collagen (Major Type I) was pepsinized, reduced by beta-mercaptoethanol, and reconstituted by glutaraldehyde. The reconstitution of the collagen increased the resistance to biological degradation by collagenase, optimized the pore size and possessed maximum biological activity for tissue regeneration. Forty-four New Zealand rabbits underwent either sham surgical procedures or partial temporomandibular joint discectomy. In animals that underwent partial discectomy, the discs were replaced by either reconstituted collagen templates or subdermal grafts. Some of the surgerized animals did not receive any type of implant or disc substitute. Gross and histological examination of the surgerized temporomandibular joints was carried out at 1-, 2-, and 3-month intervals after surgery on the selected groups of animals. Marked arthritic changes were observed after 3 months in the partially discectomized joints without implantation. In contrast, the discs, which received a reconstituted collagen template or subdermal graft exhibited regeneration and nearly normal morpology. No foreign body response was observed in experimental groups 3 months after implantation. This study demonstrated that the reconstituted collagen did as well as subdermal grafts in supporting and facilitating regeneration of the disc and the former was found to have some advantages over the latter.

Demand for traditional medicine in Taiwan: a mixed Gaussian-Poisson model approach.

Hurdle count models are used to examine the participation and consumption decisions in Chinese medicine use. Motivated by a household production model, a second censoring mechanism is introduced into existing single-hurdle models, and the resulting specification accommodates conscientious abstainers, as well as economic non-consumers, and admits excessive zeros in the sample. In contrast to previous studies that found few predictors, empirical results based on a Taiwanese national sample suggest that Western medicine is a gross substitute to Chinese medicine, and both time price and money price play more important roles than income. Insurance, lifestyle and demographics also determine the use of Chinese medicine.

Secretory origin and temporal appearance of the porcine beta-microseminoprotein (sperm motility inhibitor) in the boar reproductive system.

A specific antiserum against the porcine sperm motility inhibitor (SMI) was used in Western blotting analysis of tissue homogenates to reveal the possible origin of SMI in the boar reproductive system at different ages. The ages of the boar used were day 0, day 15, day 30, day 60, day 100, day 120, day 135, day 150, and day 210. The tissue homogenates of the day 60 and older showed immunoreaction. The results were further checked by indirect immunohistochemical staining and observed under light microscope. The SMI antigen appeared in the epithelial cells and in the lumen of the secretory ducts of the prostate gland. These results indicate that porcine SMI is synthesized only by the postnatal prostate gland. The homogenate of the prostate gland of day 100 was also used for the purification of SMI. The prostatic SMI was co-eluted with the seminal SMI in the reversed phase HPLC. Mass spectrometric analysis of the prostatic SMI revealed a molecular weight of 10,066. These results indicate that the prostatic SMI is identical to that purified from seminal plasma (Jeng et al., 1993; Biochem Biophys Res Communi 191:435-440).

Ductal adenocarcinoma of the prostate with endometrioid features in a 69-year-old man.

We describe a case of ductal adenocarcinoma of the prostate with endometrioid characteristics presenting as painless hematuria and intraurethral tumor. A 69-year-old man had intermittent painless hematuria for 2 months. The serum prostate-specific antigen concentration was elevated (22.0 ng/mL). An enlarged prostate with a necrotic tumor was noted in the right lobe of the prostate on computerized tomography and magnetic resonance imaging studies. A polypoid and worm-like tumor was found within the prostatic urethra near the verumontanum. The tumor had a distinctly papillary configuration with a focal glandular structure on microscopy. Radical prostatectomy was performed and histology of the tumor specimen revealed it to be composed of a closely packed glandular structure lined by single layers of high columnar cells with focal stratification. Frequent papillary projections of glandular epithelium and intraglandular bridging were noted, with a histopathologic appearance similar to endometrioid carcinoma of the uterus. Androgen deprivation therapy was started immediately following surgery. No evidence of recurrence or metastasis was found at follow-up 27 months postoperatively. Distinct features of ductal adenocarcinoma of the prostate include intraurethral papillary tumor close to the verumontanum, urethral obstruction, and easy bleeding of the tumor. Its more aggressive behavior than classical microacinar adenocarcinoma of the prostate makes early recognition of this type of prostatic malignancy important.

The potential of soybean foods as a chemoprevention approach for human urinary tract cancer.

Isoflavones are excreted in human urine and can be modulated by soy-rich diets. Recently, isoflavones were suggested to have protective effects against bladder cancer cells. We sought to determine the efficacy of the antitumorigenic effects of isoflavones at concentrations found in the range of human urine excretion and compare normal urothelium and bladder cancer cells for differential cytotoxicity. A total of seven human bladder cancer cell lines and an immortalized uroepithelial cell line were used to examine the effects of genistein, daidzein, and biochanin-A, either individually or as an equal-proportion mixture regimen, on cell growth, DNA synthesis, alterations of cell cycle distribution, and induction of apoptosis. The role of cyclin B1 and cdc2 kinase in cell cycle arrest was analyzed. In addition, severe combined immunodeficient mice were used to confirm the anti-cancer effects of isoflavones in vivo. Cooperative action of isoflavones was more effective in growth inhibition and apoptosis induction than any single compound. Genistein tends to cause a dose-dependent induction of G2-M cell cycle arrest and an inhibition of cdc2 kinase activity. However, both daidzein and biochanin-A directly induced apoptosis without altering cell cycle distribution. The IC50 values in non-transformed cells were higher than those in most cancer cell lines, and the IC50 of the mixture regimen was within reach of the levels observed in urine after a soy challenge. Furthermore, both genistein and combined isoflavones exhibited a significant tumor suppressor effect in vivo (P < 0.05). The results justify the potential use of soybean foods as a practical chemoprevention approach for patients with urinary tract cancer.

Alpha 1-acid glycoprotein-induced tumor necrosis factor-alpha secretion of human monocytes is enhanced by serum binding proteins and depends on protein tyrosine kinase activation.

The acute phase protein, alpha1 acid glycoprotein (AGP), stimulated human mononuclear cells as well as monocytes to secrete tumor necrosis factor-alpha (TNFalpha) which was demonstrated by ELISA, RT-PCR and functional assays. AGP-induced TNFalpha secretion of monocytes was enhanced in the presence of human plasma and inhibited by protein kinase inhibitors, indicating it is serum and tyrosine kinase dependent. The activation of tyrosine kinase in AGP-stimulated monocytes was further confirmed by immunoblotting of tyrosine phosphorylated proteins of monocytes at different time after AGP stimulation. Furthermore, several serum proteins such as C3, sCD14 and IgG were able to bind to AGP and enhanced TNFalpha secretion of human monocytes induced by AGP. Taken together, these results suggest serum proteins binding to AGP enhance its ability to stimulate human monocytes to secrete pro-inflammatory cytokines through a tyrosine kinase dependent pathway.

The dynamic responses of pro-inflammatory and anti-inflammatory cytokines of human mononuclear cells induced by uromodulin.

This study was undertaken to examine the dynamic response of human peripheral blood mononuclear cells (PBMC) in the secretion of proinflammatory and anti-inflammatory cytokines induced by uromodulin (URO). Levels of tumor necrosis factor-alpha (TNFalpha), TNF soluble receptor (sTNFRI and II), interleukin 1-beta (IL-1beta), and IL-1 receptor antagonist (IL-1Ra) in the supernatants of URO-stimulated PBMC were measured by ELISA. URO stimulated the secretion of all these cytokines in a dose dependent manner except sTNFRI. Peak levels of TNFalpha and IL-1beta were reached at 6-12 h, while 5-10 fold higher in sTNFR II and IL-1Ra levels were observed at 24-48 h after URO stimulation. URO-induced secretion of TNFalpha, IL-1beta, sTNFRII and IL-1Ra could be enhanced by human plasma. Specifically, serum proteins including C3, sCD14 and IgG not only bound to URO but also enhanced URO-induced TNFalpha secretion of PBMC. Collectively, our data suggest that URO might have dual immunomodulating effect through regulating the secretion of proinflammatory and anti-inflammatory cytokines, and that serum binding proteins might enhance this activity.

Uromodulin and Tamm-Horsfall protein induce human monocytes to secrete TNF and express tissue factor.

Effects of uromodulin (URO) and Tamm-Horsfall protein (THP), the most abundant proteins in the urine of pregnant and normal women, respectively, on the induction of TNF-alpha secretion and tissue factor (TF) expression of human monocytes were studied. THP, URO, and its fragments stimulated human mononuclear cells to proliferate and secrete TNF-alpha. The release of URO and THP-induced TNF-alpha in monocytes was dependent upon protein tyrosine kinase activation that results in tyrosine phosphorylation. URO and THP also induced TF expression of human monocytes and monocytic cell line U937 in a dose-dependent manner. TF expression was transient, reached its peak at 6 h and declined toward basal levels by 24 h. Reverse transcriptase-PCR and dot-blot analysis confirmed the induction of TF mRNA synthesis. URO and THP-induced TF expression were inhibited by actinomycin D and pentoxifylline further supporting the requirement of de novo TF mRNA synthesis. The possibility of LPS contamination of URO and THP was excluded because: 1) URO and THP-induced TF expression were inhibited by specific Ab; 2) URO was less capable of inducing TF in HUVEC as compared with LPS; 3) polymyxin B blocked the induction of Limulus clotting by LPS but not by URO and THP; 4) both LPS-sensitive (C3H/HeN) and -resistant (C3H/HeJ) mice produced little or no TNF-alpha after URO challenge. Therefore, our findings suggest that URO and THP play a significant role in the innate immunity of the urinary system and that the immunostimulatory activity of URO is potentially useful for immunotherapy.

A substance isolated from Cornus officinalis enhances the motility of human sperm.

The effects of a Chinese herb, Cornus officinalis, on the motility of human sperm was studied. An aqueous extract was prepared from the dried fruits of the herb and used in this study. The crude extract at a final concentration of 0.5 microgram/microliter in phosphate buffered saline (pH 7.4) increased sperm motility from 25.8 +/- 7.7% to 42.8 +/- 10.3% (i.e. 68% increase, n = 7), as determined by the computer-aided-sperm-analysis (CASA) method. The crude extract was fractionated by high-performance liquid chromatography (HPLC) into four fractions: C1, C2, C3 and C4. Their effects on sperm motility were further studied by CASA. Only the C4 fraction showed substantial stimulatory effects on sperm motility. At a concentration of 5 ng/microliter, C4 increased the sperm motility from 15.7 +/- 3.8% to 34.5 +/- 6.4% (i.e. 120% increase, n = 6) by CASA and from 14.9 +/- 4.3 to 28.5 +/- 8.1 (i.e. 91% increase, n = 8) by transmembrane migration ratio (TMMR) method. This result suggests that C4 is the active component in Cornus officinalis that enhances sperm motility.

Effects of alpha 1-acid glycoprotein on tissue factor expression and tumor necrosis factor secretion in human monocytes.

Activated monocytes express tissue factor (TF) and secrete tumor necrosis factor alpha (TNF alpha), which are important in the initiation of blood coagulation and inflammation. We investigated the effect of alpha 1-acid glycoprotein (alpha 1-AGP), an acute phase protein, on the induction of the expression of TF and the secretion of TNF alpha in human monocytes in vitro. The TF activity of both fresh human monocytes and human monocytic cell line U937 significantly increased in a dose-dependent manner after a 6 h incubation with human or bovine alpha 1-AGP. The activity of TF gradually tailed off after 24 h. RT-PCR and Southern blot analysis revealed that TF mRNA synthesis was induced in monocytes. Inhibition of alpha 1-AGP induced TF expression by actinomycin D (ActD) further support that de novo TF mRNA synthesis was required. The specificity of the alpha 1-AGP-induced TF activity was demonstrated by anti-alpha 1-AGP antibody inhibition. TNF alpha secretion in alpha 1-AGP stimulated monocytes was also increased; this could be blocked by pentoxifylline (PTX). The possible contamination of lipopolysaccharide (LPS) in the alpha 1-AGP was excluded by limulus amoebocyte lysate. Therefore, these results indicate that alpha 1-AGP may contribute to the cellular initiation of coagulation and inflammation by increasing TF expression and TNF alpha secretion of monocytes.

Comparative studies of types 1 and 2 herpes simplex virus infection of cultured normal keratinocytes.

AIMS: To investigate the differences in biological properties, multiplication patterns, and cytopathic effects between type 1 and type 2 herpes simplex virus (HSV) through the replication of HSV in cultured normal human keratinocytes. METHODS: Keratinocytes were obtained from surgical specimens of normal gingiva, cervix, trunk skin, and newborn foreskin. They were cultured in serum free, chemically defined, culture medium and infected with a pool of HSV collected from clinical specimens. RESULTS: The reproductive patterns of HSV type 1 (HSV-1) and HSV type 2 (HSV-2) differed from each other regardless of the anatomical source of the cultured cells. This was made evident by the dissimilarity of their growth curves and cytopathic effects. The growth curve of HSV-2 showed a more or less continuously rising titre, whereas HSV-1 titres varied substantially at different time intervals. The cytopathic effects induced by HSV-1 infection took 24 more incubation hours than those induced by HSV-2 infection to manifest. During the early stages, the cytopathic changes of the two viruses looked different. However, all cultured cells, whether cultured with HSV-1 or HSV-2, eventually became small and globular in shape. The infective titres of both HSV-1 and HSV-2 were higher in infected cultured cervix than in infected cultured normal gingiva. CONCLUSIONS: These data suggest that each serotype of HSV has its own unique replication pattern in human keratinocytes regardless of the cell origin.

Congenital depression of the fetal skull: case report and review of literature.

Congenital skull depression in babies delivered by cesarean section (C/S) is an extremely rare occurrence and its etiology is frequently unknown. Such depression may be associated with brain injury and permanent cosmetic deformity. Surgical therapy is not the only choice since cranial CT is useful for identification of intracranial damage. In our case, CT scan revealed no intracranial injury and neurological examination of the baby was normal. Thereafter, the baby was managed expectantly and spontaneous resolution of the skull depression was noted by 14 weeks of age.

Comparison of associated high-risk factors and perinatal outcome between symmetric and asymmetric fetal intrauterine growth retardation.

This study compares associated high-risk factors and perinatal outcome between 273 symmetric and 445 asymmetric infants with intrauterine growth retardation. No differences were seen in 17 obstetric, medical, and environmental-behavioral high-risk factors between symmetric and asymmetric intrauterine growth retardation. In preeclampsia, the incidence of symmetric intrauterine growth retardation is higher than that of asymmetric intrauterine growth retardation. The timing of the interaction between the high-risk factor and the stage of gestation is more important than the specific high-risk factor in determining whether symmetric or asymmetric intrauterine growth retardation is produced. On comparison of perinatal outcome between the two groups, we concluded: (1) that the onset of symmetric intrauterine growth retardation occurs much earlier in the course of pregnancy than does asymmetric intrauterine growth retardation, (2) that more symmetric than asymmetric pregnancies with intrauterine growth retardation result in preterm delivery, (3) that the neonatal morbidity rate for symmetric intrauterine growth retardation is higher than that for asymmetric intrauterine growth retardation, and (4) that term symmetric infants with intrauterine growth retardation tend to have a lower mean birth weight and a higher incidence of small placentas than term infants with asymmetric intrauterine growth retardation.

Evaluation of the equivocal test results of Treponema pallidum haemagglutination assay.

Two hundred and eighty Rapid Plasma Reagin (RPR) positive sera with an emphasis on cases with negative and borderline positive Treponema pallidum haemagglutination assay (TPHA) results were selected. Modified TPHA (M-TPHA) and fluorescent treponemal antibody absorption (FTA-abs) tests were used for comparison. One hundred and twenty five samples were TPHA negative, of which 78 and 69 cases were also negative by M-TPHA and FTA-abs, respectively. Eighty one sera negative by TPHA at a titre of 1/80 and positive at 1/40, considered to be negative according to the manufacturer's instructions, were also negative by M-TPHA (n = 11) and by FTA-abs (n = 1). Fifty borderline positive TPHA specimens gave one negative result by both M-TPHA and FTA-abs. The remaining 24 sera were positive by all three tests. Because of the high percentage of TPHA negative results among the positive RPR sera which became reactive when rechecked by the FTA-abs, it is concluded that as a confirmatory test the TPHA should be used not instead of but in addition to the FTA-abs.

Plasmacytoma of pleural origin associated with Bence Jones proteinuria.

A 33-year-old male presented with right side pleural plasmacytoma with secretion of Bence Jones (BJ) protein and deposition of amyloid. The pleural effusion contained a prominent peak of kappa-type BJ protein on the background of polyclonal gammopathy. Pleural biopsy disclosed features of plasmacytoma with amyloid deposition. A small peak of BJ protein was also present in serum electrophoretogram on the background of increased polyclonal IgG and IgA. Kidney function was impaired and urine protein ranged from 7.8 to 19.1 g/day with 92.5% bimodal BJ protein. There were no bone lesions by systemic bone survey, and repeated bone biopsies and marrow aspirations showed plasma cells of less than 5%. The clinical course worsened progressively; over the next five months the pleurisy became bilateral and peritoneal involvement supervened. This is believed to be the first report of primary plasmacytoma involving successively pleurae and peritoneum.