Synergistic Inhibition of Delayed Rectifier K+ and Voltage-Gated Na+ Currents by Artemisinin in Pituitary Tumor (GH3) Cells.

BACKGROUND: Artemisinin (ART) is an anti-malarial agent reported to influence endocrine function. METHODS: Effects of ART on ionic currents and action potentials (APs) in pituitary tumor (GH3) cells were evaluated by patch clamp techniques. RESULTS: ART inhibited the amplitude of delayed-rectifier K+ current (IK(DR)) in response to membrane depolarization and accelerated the process of current inactivation. It exerted an inhibitory effect on IK(DR) with an IC50 value of 11.2 µM and enhanced IK(DR) inactivation with a KD value of 14.7 µM. The steady-state inactivation curve of IK(DR) was shifted to hyperpolarization by 10 mV. Pretreatment of chlorotoxin (1 µM) or iloprost (100 nM) did not alter the magnitude of ART-induced inhibition of IK(DR) in GH3 cells. ART also decreased the peak amplitude of voltage-gated Na+ current (INa) with a concentration-dependent slowing in inactivation rate. Application of KMUP-1, an inhibitor of late INa, was effective at reversing ART-induced prolongation in inactivation time constant of INa. Under current-clamp recordings, ART alone reduced the amplitude of APs and prolonged the duration of APs. CONCLUSION: Under ART exposure, the inhibitory actions on both IK(DR) and INa could be a potential mechanisms through which this drug influences membrane excitability of endocrine or neuroendocrine cells appearing in vivo.

Synthesis and antitumor activity of feruloyl and caffeoyl derivatives.

We developed two efficient protocols for the synthesis of feruloyl and caffeoyl derivatives from commercial vanillin and veratraldehyde. Pharmacological activities were assessed against a panel of human cancer cell lines in vitro. Most synthesized compounds demonstrated attractive cytotoxicity. Several new compounds demonstrated significant antiproliferative and cytotoxic activities against HeLa and Bewo tumor cell lines. In particular, 5-nitro caffeic adamantyl ester showed broad spectrum of tumor inhibition in 10 cell lines, and reduced tumor weight by 36.7% in vivo when administered at a dose of 40 mg kg(-1).

A new isoprenyl-phenol-type meroterpenoid from mangrove endophytic fungus Aspergillus sp. GXNU-Y65.

Asperphenol A (1), a new isoprenyl-phenol-type meroterpenoid, was isolated from the mangrove endophytic fungus Aspergillus sp. GXNU-Y65 together with five known compounds (2-6). All structures were assigned using extensive NMR spectroscopic data and electronic circular dichroism (ECD) calculations. Compounds 1-6 were evaluated for their cytotoxic activity against A549 and T24 human cancer cell lines. Among them, compounds 1 and 5 exhibited moderate inhibitory activities against T24 cancer cell lines with the IC50 values of 26.71 and 43.50 µM, respectively.

[Identification of major components of traditional Chinese medicine Naodesheng tablet by HPLC-DAD-MS(n)].

OBJECTIVE: To identify the major components of traditional Chinese medicine Naodesheng tablet. METHODS: A HPLC-DAD-MS(n) based method was developed to analyze and identify the major components of Naodesheng tablet. Separation was performed on an Agilent Zorbax SB-C(18) column (4.6 mm X 250 mm, i.d, 5 µm) with mobile phase consisting of water with 0.05 % formic acid and acetonitrile as gradient eluent at the flow rate of 0.5 ml.min(-1). RESULTS: A total of 43 components were detected, among which 22 were identified by comparing their UV absorption profiles, the information of molecular Glucosyl puerarin weights, and structures provided by ESI-MS(n) with those of available standards and reference data, such as Safflor yellow A, 4'-O-Glucosyl puerarin, 3'-hydroxypuerarin, Genistein-8-C-apiosyl (1-6) glucoside, Puerarin, 6"-O-xylosyl puerarin, 6"-O-apiosyl puerarin, 3'-methoxy puerarin, 3'-methoxy-6"-o-xylosyl puerarin, Daidzin, Genistin, Pueroside A, Notoginsenoside R(1), Ginsenoside Re, Ginsenoside Rg1,Daidzein,Biochanin A,Ginsenoside Rb(1), Ginsenoside Rc, Ginsenoside Rb(2), Ginsenoside Rb(3), Ginsenoside Rd. CONCLUSION: The proposed method can identify the main components of Naodesheng tablet and provide information for the quality control of this medicine.

Antiparasitical efficacy of sophoraflavanone G isolated from Sophora flavescens against parasitic protozoa Ichthyophthirius multifiliis.

Ichthyophthirius multifiliis, a global distributed protozoan parasite, causes "White spot disease" and leads to serious mortality of freshwater fish in aquaculture. The present study was conducted to assess the anti-I. multifiliis efficacy of active compound isolated from Sophora flavescens. The isolated active compound was identified as sophoraflavanone G (SG) with ESI-MS and NMR. In vitro tests, SG at concentrations of 0.5 mg/L and 2 mg/L resulted in death of all theronts and tomonts, respectively; SG at concentrations of 0.125 mg/L and 0.25 mg/L notably decreased theronts infectivity (p < 0.05). Additionally, the in vivo test results showed that a cumulative delivery of SG at concentration of 2 mg/L for 7 days protected fish from I. multifiliis infection. The 96-h LC50 (median lethal concentration) and safety concentration of SG to grass carp were 46.6 mg/L and 11.3 mg/L, respectively. The present work indicated that SG was a potential safe and effectively therapeutic agent in treating I. multifiliis.

Percutaneous Endoscopic Posterior Decompression for Therapy of Spinal Cord Compression Due to Ossification of the Ligamentum Flavum: A Long-Term Follow-up.

BACKGROUND: Thoracic ossification of the ligamentum flavum (OLF) is a pathologic condition that affects ligamentation of the OLF and causes slowly progressive myeloradiculopathy. There is a trend toward performing minimally invasive surgery. In this study, we assess the long-term efficacy of percutaneous endoscopic surgical management of thoracic OLF. METHODS: Twenty patients with thoracic myelopathy due to thoracic OLF underwent percutaneous endoscopic surgical management. We investigated clinical outcomes and neurologic improvements. Magnetic resonance imaging was performed on all patients preoperatively and postoperatively, and intramedullary signal changes were evaluated. RESULTS: The operation time averaged at 170.4 minutes (range 110-320 minutes). The mean intraoperative blood loss was 29.6 mL (range 5-100 mL). Cerebrospinal fluid leakage occurred in 1 patient and healed well. The follow-up period ranged from 60 to 72 months. The mean score on the Japanese Orthopedic Association scale improved from (6.0 ± 1.41) preoperatively to (7.83 ± 1.40) at 1 month postoperatively to (9.67 ± 1.30) at the final follow-up (P < 0.05). At the final follow-up stage, 11 patients had excellent outcomes, 6 good, 2 fair, and 1 unchanged, according to the Hirabayashi recovery rate. No patient had postoperative deterioration in neurologic status. CONCLUSIONS: Percutaneous endoscopic surgical management of thoracic OLF has proven to be both safe and effective for the resection of the OLF in the thoracic spine.

The association of plasma high-density lipoprotein cholesterol levels and cirrhosis development in obese patients with chronic hepatitis B: a cohort study.

OBJECTIVE: Metabolic disorder is a common risk factor for cirrhosis in Asia, and it will increase the risk of cirrhosis in patients with Chronic hepatitis B (CHB). However, studies on the efficacy of plasma lipid markers which predict the happening and development of cirrhosis in obese CHB patients are limited. METHODS: In total, 3327 patients who were followed for more than 4 years' follow-up in the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine joined the program. Finally, 287 obese CHB patients were included in this study according to the results of metabolic tests. The data of baseline and follow-up were collected, and the association between them was analyzed. RESULTS: Based on the follow-up results, enrolled patients were divided into a group of cirrhosis (n = 146) and a group of noncirrhosis (n = 141). Plasma glucose and high-density lipoprotein cholesterol (HDL-C) levels in the noncirrhosis group (5.2 and 1.2 mmol/L, respectively) were significantly higher than that in the cirrhosis group (5.0 and 1.0 mmol/L, respectively), while the amount of total bile acid (TBA) in the cirrhosis group was lower than that in the cirrhosis group. Levels of HDL-C and total cholesterol were associated with liver function. Plasma HDL-C was an independent indicator of cirrhosis in patients with CHB. Patients with HDL-C levels less than 1.03 mmol/L had a 2.21-fold higher incidence rate of cirrhosis, and patients over 40 years old or the levels of HDL-C less than 1.03 mmol/L were more likely to generate cirrhosis. CONCLUSIONS: Plasma HDL-C was an appropriate marker in predicting cirrhosis for patients with CHB.

[Clinical report of the modified Piver class III hysterectomy on invasive cervical cancer].

OBJECTIVE: To explore the surgical extent and to improve the surgical techniques of the Piver class III hysterectomy on invasive cervical cancer, so as to reduce the urinary tract complications, shorten the surgical duration, decrease the hemorrhage and blood transfusion. METHODS: The study group, 196 cases with stages Ib and IIa carcinoma of the cervix underwent the modified Piver class III hysterectomy from June 2000 to May 2005. The control group, 176 cases of the same stages underwent the Piver class III hysterectomy between June 1994 and May 1999. The modified Piver class III hysterectomy mainly include the surgical extent and some surgical techniques as follows. The cervicovesical and vesicovaginal space are separated with assistance of electrotome. Half of the uterosacral ligaments are removed with electrotome. The tunnel of the ureters is separated and penetrated or not. The anterior leaf of the cervicovesical ligaments is removed and the uterine artery are removed at the same time, while the ureter branch from the uterine artery are preserved. When the ureters are drawn to the lateral side of the body with an "S" hook and the urocyst lateral recessus are expanded, the cardinal ligaments can be exposed and be removed of 3/4. But part of the inferior of these ligaments should be preserved. The paracolpium are resected about 2 cm, 2 - 3 cm tissue of the vagina is removed. RESULTS: Compare with the control group, the urinary tract complications of the study group were significantly reduced (51.1% versus 23.0%, P < 0.01). There were a shorter surgical duration [(132 ± 20) min], less of the hemorrhage [(322 ± 100) ml] and blood transfusion [(154 ± 79) ml] in the study group than those in the control group (all P < 0.05). While, there was no significant difference at the survival rates of 5 years between the two groups (87.8% versus 88.6%, P = 0.793). CONCLUSION: The modified Piver class III hysterectomy is effective and applicable for patients with cervical cancer.

[Expression level and effect of P2X7 receptor mRNA during osteogenic differentiation of human periodontal ligament stem cells].

PURSPOSE: To investigate the role of P2X7 receptor in osteogenic differentiation of human periodontal ligament stem cells. METHODS: Human periodontal ligament stem cells obtained from primary culture were divided into 4 groups: control group, adenosine triphosphate group, osteogenic induction group, adenosine triphosphate + osteogenic induction group. The differences of RUNX2, OCN gene expression and P2X7 receptor mRNA expression between the four groups were compared. Statistical analysis was performed with SPSS 16.0 software package. RESULTS: One week after osteogenic formation and two weeks after osteogenic formation, the expression of RUNX2 and OCN mRNA in the adenosine triphosphate + osteogenic induction group was significantly higher than that in the osteogenic induction group (P<0.05). The expression of RUNX2 and OCN mRNA in the 1 week after adenosine triphosphate + osteogenic induction fluid was significantly higher than that 2 weeks after osteogenic formation, and the difference was statistically significant (P<0.05). The expression of P2X7 receptor mRNA in the adenosine triphosphate group and the adenosine triphosphate + osteogenic induction group was significantly higher than that in the control group and the osteogenic induction group 1 week after osteogenesis and 2 weeks after osteogenesis (P<0.05). The expression of P2X7 receptor mRNA in the adenosine triphosphate group was significantly higher than that in the adenosine triphosphate + osteogenic induction group 2 weeks after osteogenesis(P<0.05). The expression of P2X7 receptor mRNA was significantly higher than that of osteogenic induction 1 week after adenosine triphosphate composition(P<0.05). CONCLUSIONS: P2X7 receptor can significantly improve the osteogenesis of periodontal ligament stem cells, and adenosine triphosphate can activate the expression of P2X7 receptor.

[Effects of Prostaglandin E2 Receptor 4 Agonist on the Expression of Stem Factors in Human CD34+ Cells].

OBJECTIVE: To investigate the effect of prostaglandin E2 recoptor 4 antagonist (EP4A) on the self-renewal ability of human CD34+ cells and its mechamism. METHODS: The peripheral blood hematopoietic stem cell of 20 healthy donors received the G-CSF-mobilization were collected, then the human CD34+ cells were sorted out by MACS microbead kit. The human CD34+ cells were treated with DMSO (control group), EP4A (EP4A group) and EP4A+EP4A antagonist (EP4A+EP4A group) for 72 hours. The differential genes and pathways related with CD34+ cell stemness were detected by Thermogram and Pathway enrichment analysis. and then the expression levels of protein and gene (ß-catenin, Nanog, Oct4, Sox2, Stat3, AKT, P38) were detected by qRT-PCR and Western blot respectively. RESULTS: EP4A could elevate the mRNA and protein expression of ß-catenin, Nanog, Oct4, Sox2, in comparison with control group, however, mRNA and protein expression of STAT3, AKT, P38 were not changed. When human CD34+ cell were cultured with EP4A+XAV939 it was found that the mRNA and protein expression of ß-catenin was downregulated, moreover the mRNA and protein expression of Nanog, Oct4, Sox2 were reduced. CONCLUSION: EP4A can upregulate stemness factors-ß-catenin, Nanog, Oct4 and Sox2 in human CD34+ cell in vitro, but not STAT3, AKT and P38.

In vivo antibacterial activity of vertilmicin, a new aminoglycoside antibiotic.

Vertilmicin is a novel aminoglycoside antibiotic with potent activity against gram-negative and -positive bacteria in vitro. In this study, we further evaluated the efficacy of vertilmicin in vivo in systemic and local infection animal models. We demonstrated that vertilmicin had relatively high and broad-spectrum activities against mouse systemic infections caused by Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Enterococcus faecalis. The 50% effective doses of subcutaneously administered vertilmicin were 0.63 to 0.82 mg/kg, 0.18 to 0.29 mg/kg, 0.25 to 0.99 mg/kg, and 4.35 to 7.11 mg/kg against E. coli, K. pneumoniae, S. aureus, and E. faecalis infections, respectively. The therapeutic efficacy of vertilmicin was generally similar to that of netimicin, better than that of gentamicin in all the isolates tested, and better than that of verdamicin against E. coli 9612 and E. faecalis HH22 infections. The therapeutic efficacy of vertilmicin was further confirmed in local infection models of rabbit skin burn infection and mouse ascending urinary tract infection.

Class II radical hysterectomy in low-risk IB squamous cell carcinoma of cervix: a safe and effective option.

UNLABELLED: The aim of this study was to determine the outcome of class II radical hysterectomy for the treatment of a subset of patients with early cervical cancer. METHOD: From September 1995 to September 2003, a total of 480 patients whose conditions were diagnosed with squamous carcinoma of the cervix with a tumor size of no greater than 2 cm and a pathological grade 1 to 2 was enrolled in a cohort study to compare class II and class III radical hysterectomy, with 240 patients in each group. Disease-free survival, overall survival, pattern of recurrences, and morbidity were the end points of observation for this study. RESULTS: The class II surgery group had 100.0% overall and 98.33% disease-free actuarial 5-year survival rate, whereas the class III surgery group had 100.0% overall and 97.92% disease-free actuarial 5-year survival rate (P = 0.736). Recurrence rate (2.92% in class II vs 2.50% in class III) was not significantly different in the 2 groups (P = 0.779). The operating time, postoperative length of hospital stay, and estimated blood loss at surgery were significantly lower in the group of patients who underwent class II hysterectomy (P = 0.0001, P = 0.0001, and P = 0.001, respectively). The postoperative complications were also significantly lower in patients who received class II hysterectomy. CONCLUSIONS: In treating a subset of patients with tumor size no greater than 2 cm and grade 1 to 2 squamous cell cancer, class II and class III radical hysterectomy are equally effective, but the former has far less complications than the latter.

Plasma sFlt-1-to-PlGF ratio is correlated with inflammatory but not with oxidative stress in Chinese preeclamptic women.

PURPOSE: Considerable interest has been focused on angiogenic factors and angiogenic imbalance in the field of pre-eclampsia (PE), owing to its gaining role in the development of PE. This study was addressed to investigate the associations of sFlt-1-to-PlGF plasma ratios with oxidative stress assessed by the level of 8-isoprostane, and inflammation measured by the level of high-sensitive C-reactive protein (hs-CRP), and adipocytokines. METHODS: A total of 83 patients with PE including 47 mild PE (MPE) and 36 severe PE (SPE) and 50 age-matched normotensive subjects in the third trimester of pregnancy were examined. Measurements included body mass index (BMI), systolic and diastolic blood pressure (BP) levels, plasma concentrations of hs-CRP, 8-isoprostane, adiponectin, and leptin. RESULTS: Subjects with PE had higher levels of sFlt-1/PlGF (P < 0.01), hs-CRP (P < 0.01), 8-isoprostane and leptin (both P < 0.01) and lower adiponectin (P < 0.01) than did normotensive control subjects. Significant positive correlations were found between plasma sFlt-1/PlGF and hs-CRP (r = 0.437, P < 0.01) or leptin (r = 0.656, P < 0.01). A weak inverse correlation emerged between sFlt-1/PlGF and adiponectin (r = -0.306, P < 0.01). When a multiple regression analysis was performed, with sFlt-1/PlGF as a dependent variable and all the other parameters as independent variables, sFlt-1/PlGF maintain a significant relationship with leptin (beta = 0.219, P < 0.05) and with hs-CRP (beta = 0.295, P < 0.01) as well as with systolic BP(beta = 0.446, P < 0.05). CONCLUSIONS: In Chinese preeclamptic women, plasma sFlt-1-to-PlGF ratio is correlated with inflammatory and adipocytokines but not with oxidative stress.

In vitro antibacterial activity of vertilmicin and its susceptibility to modifications by the recombinant AAC6'-APH2'' enzyme.

Vertilmicin is a new semisynthetic aminoglycoside with a structure similar to that of netilmicin except for a methyl group at the C-6' position. In the present study, the in vitro antibacterial activity of vertilmicin was studied, and its susceptibility to modifications by the recombinant aminoglycoside bifunctional modifying enzyme AAC(6')-APH(2'') was compared with those of verdamicin and netilmicin. A total of 1,185 clinical isolates collected from hospitals in Beijing between 2000 and 2001 were subjected to the in vitro antibacterial activity evaluations, including MIC, minimum bactericidal concentration (MBC), and time-kill curve tests. The MICs were evaluated in non-gentamicin-resistant (gentamicin-susceptible and gentamicin-intermediate) strains and gentamicin-resistant strains, respectively. For most of the non-gentamicin-resistant bacteria (except for the isolates of Pseudomonas spp.), the MIC(90)s of vertilmicin were in the range of 0.5 to 8 microg/ml, comparable to those of the reference aminoglycosides. For the gentamicin-resistant isolates, the three semisynthetic aminoglycosides (vertilmicin, netilmicin, and amikacin) demonstrated low MIC(50)s and/or MIC(90)s, as well as high percent susceptibility values. Among the study drugs, vertilmicin showed the lowest MIC(90)s, 16 microg/ml, for the gram-positive gentamicin-resistant isolates of Staphylococcus aureus and Staphylococcus epidermidis. Meanwhile, vertilmicin was a potent bactericidal agent, with MBC/MIC ratios in the range of 1 to 2 for Escherichia coli, Klebsiella pneumoniae, and S. aureus and 1 to 4 for S. epidermidis. The time-kill curve determination further demonstrated that this effect was rapid and concentration dependent. In evaluations of susceptibility to modifications by the recombinant AAC(6')-APH(2'') with maximum rate of metabolism/K(m) measurements, vertilmicin exhibited susceptibilities to both acetylation and phosphorylation lower than those of netilmicin and verdamicin.

In vitro activity of recombinant lysostaphin against Staphylococcus aureus isolates from hospitals in Beijing, China.

Lysostaphin is a glycylglycine endopeptidase. It cleaves the pentaglycine cross-bridge structure unique to the staphylococcal cell wall and is considered to be a potential drug for Staphylococcus aureus. In the present study, the in vitro activity of recombinant lysostaphin was investigated in 257 S. aureus isolates collected from hospital patients in Beijing, China, by determination of MIC and minimum bactericidal concentration (MBC) and a time-kill curve test. An agar dilution method was used for MIC determination in all of the isolates and a macrobroth dilution method was employed to verify MIC values for a subset of the isolates. All of the S. aureus strains were sensitive to the recombinant lysostaphin with MICs ranging from 0.03 to 2 microg ml(-1) in the agar dilution assay. The antibacterial activity of lysostaphin was greater than that of vancomycin and other reference agents. For most of the isolates, the MICs from the agar dilution method were higher than those from the broth dilution method. The MBCs of lysostaphin in the test isolates were between 1- and 8-fold higher than their MIC values. Bactericidal activity (>99.9 % reduction) was observed after 2 h exposure of the isolates to lysostaphin at concentrations of > or =0.5 MIC. Lysostaphin showed a rapid bactericidal activity against the test strains of meticillin-susceptible S. aureus and meticillin-resistant S. aureus. Its activity at > or =0.5 MIC was sustained for at least 6 h. These results will be informative for the clinical application and evaluation of lysostaphin.

Transforaminal Percutaneous Endoscopic Discectomy and Foraminoplasty after Lumbar Spinal Fusion Surgery.

BACKGROUND: The most common causes of pain following lumbar spinal fusions are residual herniation, or foraminal fibrosis and foraminal stenosis that is ignored, untreated, or undertreated. The original surgeon may advise his patient that nothing more can be done in his opinion that the nerve was visually decompressed by the original surgery. Post-operative imaging or electrophysiological assessment may be inadequate to explain all the reasons for residual or recurrent symptoms. Treatment of failed lumbar spinal fusions by repeat traditional open revision surgery usually incorporates more extensive decompression causing increased instability and back pain. The authors, having limited their practice to endoscopic surgery over the last 10 years, report on their experience gained during that period to relieve pain by transforaminal percutaneous endoscopic revision of lumbar spinal fusions. OBJECTIVE: To assess the effectiveness of transforaminal percutaneous endoscopic discectomy and foraminoplasty in patients with pain after lumbar spinal fusion. STUDY DESIGN: Retrospective study. SETTING: Inpatient surgery center. METHODS: Sixteen consecutive patients with pain after lumbar spinal fusions presenting with back and leg pain that had supporting imaging diagnosis of foraminal stenosis and/or residual/recurrent disc herniation, or whose pain complaint was supported by relief from diagnostic and therapeutic injections, were offered percutaneous transforaminal endoscopic discectomy and foraminoplasty over a repeat open procedure. Each patient sought consultation following a transient successful, partially successful or unsuccessful open lumbar spinal fusions treatment for disc herniation or spinal stenosis. Endoscopic foraminoplasty was also performed to either decompress the bony foramen in the case of foraminal stenosis, or to allow for endoscopic visual examination of the affected traversing and exiting nerve roots in the axilla. The average follow-up time was 30.3 months, minimum 12 months. Outcome data at each visit included MacNab criteria, visual analog scale (VAS), and Oswestry Disability Index (ODI). RESULTS: The average leg VAS improved from 9.1 ± 2.0 to 2.0 ± 0.8 (P < 0.005). Ten patients had excellent outcomes, 5 had good outcomes, one had a fair outcome, and none had poor outcomes, according to the MacNab criteria. Fifteen of 16 patients had excellent or good outcomes, for an overall success rate of 93.7%. No patients required reoperation. There were no incidental durotomies, infections, vascular, or visceral injuries. There was one complication, a case of leg numbness caused by dorsal root ganglion injury. The numbness improved after 2 weeks. After 3 months, physical exam showed that the total area of numbness in the legs had decreased. At last follow-up, the patient had no pain, and only a few areas with numbness remained that did not affect the patient's activities of daily living. The patient was relieved to be able to avoid open decompression. LIMITATIONS: This is a retrospective study. CONCLUSIONS: The transforaminal endoscopic approach is effective for patients with back or leg pain after lumbar spinal fusions due to residual/recurrent nucleus pulposus and foraminal stenosis. Failed initial index surgery may involve failure to recognize patho-anatomy in the axilla of the foramen housing the traversing and the exiting nerve. The transforaminal endoscopic approach effectively decompresses the foramen and does not further destabilize the spine needing stabilization. It also avoids going through the previous surgical site. KEY WORDS: Full-endoscopic, foraminal stenosis, recurrent herniation, surgical treatment, fusion.

[Prostaglandin E2 Receptor 4 Agonist Promotes Human CD34+ Cell Proliferation in vitro by Activating Wnt/ß-Catenin Signaling Pathway].

OBJECTIVE: To investigate the potential signaling pathway that regulates the proliferation of human CD34+ cells stimulated by prostaglandin E2 receptor 4 agonist (EP4A) in vitro. METHODS: Twenty samples of peripheral blood containing stem cells were collected from the G-CSF mobilized healthy donors in our department of hematology. Human CD34+ cells were isolated by magnetic activated cell sorting (MACS) microbeads kit. The Cell Counting Kit-8 (CCK8) assay was used to determine the optimal concentration and time of EP4A to promote human CD34+ cell proliferation in vitro. Under the optimal condition, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect mRNA level of ß-catenin, and Western blot was used to assay protein expression of ß-catenin and P-GSK-3ß in human CD34+ cells treated with EP4A. RESULTS: Culturing with 10 µmol/L EP4A for 72 h, it was found that EP4A promoted human CD34+ cell proliferation significantly, and the proliferation rate of human CD34+ cells was 1.36 times higher than that of the control(P=0.002). Under the optimal condition, it was also found that EP4A enhanced the ß-catenin expression at both mRNA and protein levels, and up-regulated phosphorylation of GSK-3ß in human CD34+ cells, but these effects could be inhibited by the EP4A antagonist EP4AA. CONCLUSION: EP4A can enhance human CD34+ cell proliferation in vitro by activating Wnt/ß-catenin signaling pathway.

[Association between expression of survivin and high-risk human papillomavirus infection in cervical cancer and precancerous tissues].

OBJECTIVE: To investigate the expression of survivin and its association with high-risk human papillomavirus (HPV) infection in cervical cancer and precancerous tissues. METHODS: In 76 patients with cervical intraepithelial neoplasia (CIN) or cervical carcinoma, the expression of survivin was detected by immunohistochemistry and the infection rates of high-risk HPV including HPV16, 18, 31, 33, 52, and 58 were assayed by touch-down PCR, with 10 cases of normal cervical specimens serving as the control. RESULTS: Survivin expression tended to increase along with the tumor progression (P<0.05), and was associated with the histological grade of invasive carcinoma (P<0.05) but not with the patients' age, clinical stage or histological classification (P>0.05). The positivity rate of high-risk HPV increased only in association with tumor progression (P<0.05) but not with the patients' age, clinical stage or histological classification (P>0.05). Positive correlation between the expression of survivin and high-risk HPV infection was observed in cervical carcinoma. CONCLUSION: Survivin might play an important role in the occurrence and development of cervical carcinoma in coordination with high-risk HPV.

[Effects of hepatocyte growth factor on IL-1alpha triggered tubular epithelial-myofibroblast transdifferentiation and fibronectin secretion in vitro].

OBJECTIVE: To observe the effects of hepatocyte growth factor (HGF) on tubular epithelial-myofibroblast transdifferentiation (TEMT) triggered by IL-1alpha and the fibronectin secretion of TEMT. METHODS: The normal rat kidney tubular epithelial cell line (NRK52E) was cultured for six days on plastic or collagen type I-coated plates in the presence or absence of HGF or IL-1alpha. The morphology of transdifferentiation tubular cells was observed by scanning electron microscopy (SEM) and phase-contrast microscopy. The number of alpha-SMA+ cells, the percentage of alpha-SMA+ cells and the mean channel fluorescence (MCF) were assessed by immunohistochemistry and flowcytometry. The level of fibronectin in supernatant was measured by ELISA. RESULTS: The NRK52E cells triggered by IL-1alpha became fibroblast-like morphologically, and strong alpha-SMA immunostaining of those cells was seen. The level of FN in the culture supernatant, the percentage of alpha-SMA+ cells and the MCF of cells triggered by IL-1alpha were obviously higher than those of blank control group (P<0.05). In the groups with IL-1alpha and different doses of HGF, the transdifferentiation of NRK52E cells was inhibited. With the increase of HGF dose, the percentage of alpha-SMA+ cells and the level of FN showed a tendency to decrease. There was no significant difference between the groups treated with only HGF at different dose levels and the blank control group (P>0.05). CONCLUSION: IL-1alpha can induce tubular epithelial cell to transdifferentiate to myofibroblast and increase the secretion of FN. These results suggest that TEMT may play an important role in the pathogenesis of renal fibrosis. HGF could block the transdifferentiation of tubular epithelial cell and inhibit the secretion of FN. These would provide a novel therapeutic strategy for the treatment of renal interstitial fibrosis and end stage renal disease.