[Comparison of maternal-fetal outcomes between laparoscopic cervicoisthmic cerclage and McDonald cerclage in women with cervical insufficiency].

Objective: To compare the maternal and fetal outcomes of women with cervical insufficiency (CI) undergoing McDonald cerclage (MC) and laparoscopic cervicoisthmic cerclage (LCC), so as to provide evidence for the selection of cerclage methods. Methods: A retrospective trial was carried out in the First Affiliated Hospital of Sun Yat-sen University from January 2010 to December 2020. A total of 221 women who underwent the prophylactic cerclage were divided into MC group (n=54), LCC with MC history group (n=28) and LCC without MC history group (n=129) by the mode of operation and whether the pregnant women who underwent LCC had MC history. General clinical data, pregnancy complications and pregnancy outcomes were compared between the three groups. Results: (1) General clinical data: the proportion of women accepted cervical cerclage during pregnancy in MC group, LCC with MC history group and LCC without MC history group were 100.0% (54/54), 7.1% (2/28) and 27.1% (35/129), respectively (P<0.001). The indications of the three groups showed statistical significance (P=0.003), and the main indication was the history of abortion in the second and third trimester [75.9% (41/54) vs 89.3% (25/28) vs 84.5% (109/129)]. (2) Pregnancy complications: the incidence of abnormal fetal position [7.8% (4/51) vs 17.4% (4/23) vs 19.8% (24/121)], placenta accrete [5.9% (3/51) vs 13.0% (3/23) vs 11.6% (14/121)], uterine rupture [0 vs 4.3% (1/23) vs 5.8% (7/121)] in the MC group were all lower than those in LCC with MC history and LCC without MC history groups. However, there were no statistical significances (all P>0.05). Intrauterine inflammation or chorioamnionitis [15.7% (8/51) vs 0 vs 0.8% (1/121)] and premature rupture of membrane [23.5% (12/51) vs 4.3% (1/23) vs 0] were both significantly higher in MC group than those in LCC with MC history and LCC without MC history groups (all P<0.001). (3) Pregnancy outcomes: the cesarean section rate was significantly lower in MC group (41.2%, 21/51) than that in LCC with MC history group (100.0%, 23/23) and LCC without MC history group (100.0%, 121/121; P<0.001). MC group was associated with lower expenditure than LCC with MC history and LCC without MC history groups (12 169 vs 26 438 vs 27 783 yuan, P<0.001). The success rates of live birth cerclage did not differ significantly in MC (94.4%, 51/54), LCC with MC history (82.1%, 23/28) and LCC without MC history (93.8%, 121/129) groups (χ2=5.649, P=0.059). There was no significant difference in neonatal intensive care unit occupancy, neonatal birth weight and neonatal asphyxia between the three groups (all P>0.05). Conclusions: Both LCC and MC are the treatment choice for women with CI, which may get similar liver birth. However, MC has the advantages of low cesarean section rate, economical and easy operation. Therefore, MC is recommended as the first choice for CI patients, and LCC is for women with failed MC.

A nomogram for predicting overall survival in patients with type II endometrial carcinoma: a retrospective analysis and multicenter validation study.

OBJECTIVE: Type II endometrial cancer (EC) is associated with high risk of metastasis and poor prognosis. We aimed to develop a nomogram for predicting survival probability in patients with type II EC. PATIENTS AND METHODS: Data from a total of 4,117 patients with confirmed type II EC were pulled from the Surveillance, Epidemiology, and End Results (SEER) database, and were randomly divided into a training set and an internal verification set. A nomogram was constructed based on the training set. The concordance index (C-index), area under the ROC curve, and calibration plots were used to evaluate the identification and calibration of the nomogram. The SEER internal validation set and the Chinese multicenter data set (74 patients) were used to verify discriminations and corrections of the model. RESULTS: Multivariate analysis indicated that age, marital status, tumor size, T stage, N stage, M stage, surgery, radiotherapy, and chemotherapy were independent factors affecting the prognosis of type II EC patients (p<0.001). The corresponding nomogram has showed excellent calibration and discrimination (C-index [95% CI], 0.752 [0.738-0.766]). The model was verified in the internal verification set (0.760 [0.739-0.781]) and the Chinese multicenter set (0.784 [0.607-0.961]). In addition, the AUC further confirmed the accuracy of the nomogram in predicting survival. The calibration curve of OS within 5 years confirmed good calibration of the nomogram. CONCLUSIONS: This model and the corresponding risk classification system may provide useful tools for clinicians to evaluate the long-term prognosis of patients and carry out personalized clinical evaluation.

[Analysis of clinical features and risk factors of necrotizing pneumonia in children].

OBJECTIVE: To investigate the clinical characteristics and risk factor analysis of necrotizing pneumonia in children. METHODS: A retrospective study was used to analyze the case data of 218 children with severe pneumonia hospitalized in the Department of Respiratory Medicine, Children's Hospital of Capital Institute of Pediatrics from January 2016 to January 2020, and they were divided into 96 cases in the necrotizing pneumonia group (NP group) and 122 cases in the non-necrotizing pneumonia group (NNP group) according to whether necrosis of the lung occurred. The differences in clinical characteristics (malnutrition, fever duration, hospitalization time, imaging performance, treatment and regression follow-up), laboratory tests [leukocytes, neutrophil ratio, platelet (PLT), C-reactive protein (CRP), procalcitonin (PCT), D-dimer, and lactate dehydrogenase (LDH)] and bronchoscopic performance between the two groups were compared, and Logistic regression analysis of clinical risk factors associated with necrotizing pneumonia was performed to further determine the maximum diagnostic value of each index by subject operating characteristic curve (ROC). The critical value of each index was further determined by the ROC. RESULTS: The differences in age, gender, pathogenic classification, and bronchoscopic presentation between the two groups of children were not statistically significant (P>0.05); whereas the imaging uptake time of the children in the NP group was higher than that in the NNP group (P < 0.05). The differences in malnutrition, fever duration, length of stay, white blood cell count, neutrophil ratio, CRP, PCT, and D-dimer were statistically significant between the two groups (P < 0.05). The imaging uptake time was lower in children under 6 years of age than in those over 6 years of age, and the imaging uptake time for bronchoalveolar lavage within 10 d of disease duration was lower than that for those over 10 d; the imaging uptake time was significantly longer in the mixed infection group than that in the single pathogen infection group. Logistic regression analysis of the two groups revealed that the duration of fever, hospital stay, CRP, PCT, and D-dimer were risk factors for secondary pulmonary necrosis (P < 0.001, P < 0.001, P < 0.001, P=0.013, P=0.001, respectively). The ROC curves for fever duration, CRP, PCT, and D-dimer were plotted and found to have diagnostic value for predicting the occurrence of pulmonary necrosis when fever duration >11.5 d, CRP >48.35 mg/L, and D-dimer > 4.25 mg/L [area under ROC curve (AUC)=0.909, 0.836, and 0.747, all P < 0.001]. CONCLUSION: Children with necrotizing pneumonia have a longer heat course and hospital stay, and the imaging uptake time of mixed pathogenic infections is significantly longer than that of single pathogenic infections. Children with necrotizing pneumonia under 6 years of age have more advantageous efficacy of electronic bronchoscopic alveolar lavage within 10 d of disease duration compared with children in the group over 6 years of age and children in the group with disease duration >10 d. Inflammatory indexes CRP, PCT, and D-dimer are significantly higher. The heat course, CRP, PCT, and D-dimer are risk factors for secondary lung necrosis in severe pneumonia. Heat course >11.5 d, CRP >48.35 mg/L, and D-dimer >4.25 mg/L have high predictive value for the diagnosis of necrotizing pneumonia.

[Application of indocyanine green fluorescence visualization in surgical resection of abdominal wall endometriosis].

Objective: To investigate the feasibility, effectiveness and safety of indocyanine green (ICG) navigation in the surgical resection of abdominal wall endometriosis (AWE). Methods: Seven women undergoing surgery for AWE in First Affiliated Hospital of Sun Yat-sen University (from July 1, 2021 to October 1, 2021) were collected. After exposure of the focus, ICG were used intravenously (0.25 mg/kg) as fluorescent dye for the intraoperative evaluation of AWE vascularization. Resection of the AWE was guided by direct visualization of the focus under standard laparoscopy with a near-infrared (NIR) camera head. Surgical margin around the AWE (3, 6, 9 and 12 point) and the margin under the focus were obtained for postoperative pathological examination of endometriosis. Time from injection to fluorescence visualization, the proportion of fluorescence visualization, time of fully resection of AWE, side effects related to the use of ICG, perioperative complications as well as the pathological result of the surgical margins were recorded. Results: ICG fluorescence of the AWE were seen in 5 patients (5/7). The mean time from injection to fluorescence visualization was (46.7±9.8) s. The mean time of fully resection of AWE was (16.4±7.0) minutes. There were no side effects related to the use of ICG. The rate of class-A wound healing was 7/7. All of the surgical margins were confirmed endometriosis-negative by postoperative pathological examination. Conclusion: ICG fluorescence visualization could conduct accurate resection of AWE, which is clinically safe and effective.

[The application value of multi-material artifact reduction technique in maxillofacial CT images of patients with metallic dental implants].

Objective: To investigate the effect of multi-material artifact reduction (MMAR) technique on the maxillofacial CT images of patients with metallic dental implants. Methods: The data of 30 patients with metal denture implants who underwent maxillofacial wide-detector CT scans in the Department of Radiology of the First Affiliated Hospital of Zhengzhou University from March 2019 to February 2020 were retrospectively collected. The images of the lesion sites of patients were all affected by the metal implants artifacts. The 120 kV-like, virtual monoenergetic images (VMI) (70, 90, 110 keV), VMI (70, 90, 110 keV)+MMAR images were reconstructed at venous phase by energy spectrum scanning. A total of 7 image sequences were obtained for each patient. The CT value and noise value of the lesion and the longus capitis muscle on the same plane were measured, and the artifact index (AI) of lesion was calculated.Friedman test and Wilcoxon test were used to evaluate image quality objectively and subjectively. Results: There were significant differences in SD and AI among the 7 groups (χ2=133.800,P<0.001;χ2=92.147,P<0.001). The SD[10 (8, 12)] and AI [6 (5, 9)]of VMI 110 keV+M images were the lowest, but there were no significant differences in AI between VMI 110 keV+M and VMI 90 keV+M image (P= 0.271). The SD and AI of VMI+M group were lower than those in VMI group (all P<0.05). There was significant difference in image score among the 7 groups (χ 2 = 151.199, P<0.001). The image scores of VMI (70, 90, 110 keV)+M group were higher than those of VMI group (Z=-4.583, P = 0.000; Z=-3.742, P = 0.000; Z =-2.449, P = 0.014). The subjective score of VMI 90 keV+M [4 (3, 4)] was the highest, higher than those of other 6 groups (all P<0.05), in 26 out of 30 cases, lesions displayed clear contrast, and the artifacts eliminated completely. Conclusion: The MMAR technique of wide-detector CT might effectively remove the metallic dental implants artifacts and improve the image quality. VMI 90 keV+MMAR images could achieve the best artifact removal effect and good tissue contrast.

NLRP3 inflammasome and endoplasmic reticulum stress in the epileptogenic zone in temporal lobe epilepsy: molecular insights into their interdependence.

AIM: Nod-like receptor protein 3 (NLRP3) inflammasome-mediated inflammation has emerged as a contributor to epileptogenesis. Endoplasmic reticulum stress (ERS) plays an important role in epilepsy-induced neurodegeneration. NLRP3 activation and ERS reactions share the same induction factors, suggesting that these processes may be interdependent. However, the correlation between NLRP3 and ERS in TLE has not been confirmed. METHODS: The expression patterns of NLRP3 inflammasome and ERS-related markers in the temporal neocortices of TLE patients were investigated by western blotting, immunohistochemistry and immunofluorescent labelling. Correlations between the protein levels of NLRP3 and the expression of ERS-related markers were assessed using Spearman's rank correlation test. To observe the relationship between the NLRP3 inflammasome and ERS, inhibitors were used in a status epilepticus (SE) model. RESULTS: Our results show that NLRP3 inflammasome components and ERS-related markers were upregulated in the temporal neocortices of TLE patients, and were mainly localized to neurons, astrocytes and microglia. We found a positive correlation between the protein levels of NLRP3 and the expression of ERS-related markers in the temporal neocortices of 20 TLE patients. Furthermore, after blocking the NLRP3 inflammasome with MCC950, the expression of ERS-related markers was markedly decreased in the hippocampi of SE mice. Moreover, TUDCA, a specific ERS inhibitor, also reduced the expression of NLRP3 components in the hippocampus under SE conditions. CONCLUSION: Taken together, our data reveal the interdependence of the NLRP3 inflammasome and ERS in the epileptogenic zone of TLE patients and in the hippocampi of mice in the early post-SE phase.

[Water sorption and hygroscopic changes of five core buildup composite resins].

Objective: To evaluate water sorption and hygroscopic dimensional changes of five core buildup composite resins, and to provide references for material modification on water sorption. Methods: Five commercial core buildup materials (group A: Smart Dentin Replacement(TM); group B: NanoFil; group C: ParaCore(TM); group D: LuxaCore Z; group E: EmbraceCore(TM) Resin Cement) were fabricated to disk-shaped specimens: (15.0±0.1) mm diameter, (2.0±0.1) mm thickness (n=10). Specimens were thoroughly irradiated with curing lights. The initial mass in air was recorded, and the initial mass in deionized water was recorded. Five specimens of each group were immersed in deionized water for 28 d. They were weighed as a function of different immersion time (1, 2, 3, 4, 5, 6, 7, 14, 28 d). The mass in air was recorded, and the displayed mass in deionized water was recorded. Archimedes' principle was applied to calculate the dimensional changes. The other five specimens of each group were stored in artificial saliva and were tested by the same methods. Results: All specimens gained weight and hygroscopic changes during 28 d immersion. When stored in deionized water, the apparent mass change of group A [(10.6±0.9) µg/mm(3)] and the dimensional change of group A [(0.39±0.10)%] were the lowest, while the mass change of group E [(48.0±0.2) µg/mm(3)] and the dimensional change of group E [(3.16±0.13)%] were the highest (P<0.05). In artificial saliva, the lowest apparent mass change was found in group A [(11.8±1.0) µg/mm(3)] while the highest change was found in group E [(47.4±3.5) µg/mm(3)] (P<0.05). The lowest dimensional change was found in group C [(0.37±0.09)%] and the highest was found in group E [(3.07±0.19)%] in artificial saliva (P<0.05). Conclusions: Water sorption and dimensional changes vary in immersion fluids with different osmotic pressure. Water sorption and dimensional changes of core buildup composite resins are highly correlated with test materials in both of the deionized water and artificial saliva.

MiRNA-93 functions as an oncogene in glioma by directly targeting RBL2.

OBJECTIVE: Glioma is a tumor of the brain. Although the clinical regimens and surgical techniques for glioma have improved, therapies of advanced glioma remain challenging, carrying dismal overall survival and therapeutic success rates. Evidence has shown that miRNAs played important roles in glioma development. The current study aimed at investigating the function of a novel cancerogenic miRNA, miR-93, in glioma progression by investigating the expression and mechanism of it. PATIENTS AND METHODS: qRT-PCR was conducted to assess the miR-93 expression and the mRNA expression of target gene in glioma tissues and cells. The invasion and migration abilities of the glioma cells were determined by transwell assays. Luciferase reporter assay was performed to confirm the target of miR-93. RESULTS: The results indicated that miR-93 expression in glioma tissues and cells was increased significantly than that in normal brain tissues and cells. Furthermore, miR-93 promoted glioma cell migration and invasion. RBL2 was recognized as a direct target of miR-93 in glioma cells, and overexpression of RBL2 could reverse the stimulative effect of miR-93 in glioma cell. CONCLUSIONS: The above findings suggested that miR-93 together with RBL2 could be diagnostic targets and novel prognostic markers for glioma.

[Analysis on influencing factors of prognosis of patients with acute organophosphorus pesticide poisoning].

OBJECTIVE: To analyze the relationship between risk factors and prognosis of acute organophosphorus pesticide poisoning (AOPP). METHODS: The clinical data including APACHEⅡ, D-dimer, C-reactive protein, procalcitonin, lactic acid of the 67 cases of acute organophosphorus pesticide poisoning which respectively divided into survival group and death group by the outcome were collected. The independent influcing factors were got by logistic regression analysis. RESULTS: The analysis showed that APACHEⅡ, D-dimer、C-reactive protein and Procalcitonin were influencing factors to evaluate prognosis of AOPP (P<0.05) .Meanwhile, APACHEⅡ and CRP were the independent influencing factors to evaluate prognosis of AOPP (P<0.05). APACHEⅡ26was the optimum thresholds to acess the prognosis of AOPP and its Youden index was largest. CONCLUSION: APACHEⅡ and CRP played an important role in the assessment of prognosis on AOPP. When APACHEⅡwas more than 26, it suggested the patient of AOPP will have a bad prognosis.

Geniposide promotes beta-cell regeneration and survival through regulating ß-catenin/TCF7L2 pathway.

T-cell factor 7-like 2 (TCF7L2) is an important transcription factor of Wnt/ß-catenin signaling, which has critical roles in ß-cell survival and regeneration. In preliminary screening assay, we found geniposide, a naturally occurring compound, was able to increase TCF7L2 mRNA level in Min6 cells. Here we aimed to investigate the role of geniposide in ß-cell and underlying mechanism involved. Geniposide was found to promote ß-cell survival by increasing ß-cell proliferation and decreasing ß-cell apoptosis in cultured mouse islets after challenge with diabetic stimuli. Geniposide protected ß-cell through activating Wnt signaling, enhanced expressions of TCF7L2 and GLP-1R, activated AKT, inhibited GSK3ß activity, and promoted ß-catenin nuclear translocation. The protective effect of geniposide was remarkably suppressed by siRNAs against ß-catenin, or by ICG001 (ß-catenin/TCF-mediated transcription inhibitor). Moreover, geniposide promoted ß-cell regeneration in vivo to normalize blood glucose in high-fat diet and db/db mice. Increased ß-cell proliferation was observed in pancreatic sections of geniposide-treated diabetic mice. Most importantly, geniposide triggered small islet-like cell clusters formation as a result of ß-cell neogenesis from ductal epithelium, which was well correlated with the increase in TCF7L2 expression. In exocrine cells isolated from mouse pancreas, geniposide could induce duct cell differentiation through upregulating TCF7L2 expression and activating JAK2/STAT3 pathway. Taken together, we identified a novel role of geniposide in promoting ß-cell survival and regeneration by mechanisms involving the activation of ß-catenin/TCF7L2 signaling. Our finding highlights the potential value of geniposide as a possible treatment for type 2 diabetes.

Dynamically controlled resonance fluorescence spectra from a doubly dressed single InGaAs quantum dot.

We report the first experimental demonstration of the interference-induced spectral line elimination predicted by Zhu and Scully [Phys. Rev. Lett. 76, 388 (1996)] and Ficek and Rudolph [Phys. Rev. A 60, R4245 (1999)]. We drive an exciton transition of a self-assembled quantum dot in order to realize a two-level system exposed to a bichromatic laser field and observe the nearly complete elimination of the resonance fluorescence spectral line at the driving laser frequency. This is caused by quantum interference between coupled transitions among the doubly dressed excitonic states, without population trapping. We also demonstrate a multiphoton ac Stark effect with shifted subharmonic resonances and dynamical modifications of resonance fluorescence spectra by using double dressing.

Spectrum and features of congenital heart disease in Xi'an, China as detected using fetal echocardiography.

This study aimed to investigate the spectrum and features of congenital heart disease (CHD) in Xi'an, China using fetal echocardiography. All pregnant women referred for fetal echocardiography underwent a systematic fetal echocardiographic examination. Each case of complex defects was diagnosed according to the predominant pathophysiology, and the overall frequency of each defect was recorded and classified according to its location in the fetal heart. CHD was diagnosed in 195 fetuses. The top 5 types of CHD were, in order, single ventricle (15.9%, 31/195), atrioventricular septal defect (12.3%, 24/195), ventricular septal defect (VSD) (11.8%, 23/195), tetralogy of Fallot (10.8%, 21/195), and double-outlet right ventricle (8.2%, 16/195). The 195 cases of CHD comprised 316 defects in total. The most common defect was ventricular malformation (40.5%, 128/316), followed by great artery anomalies (38.0%, 120/316), endocardial cushion abnormalities (11.7%, 37/316), atrial abnormalities (6.6%, 21/316), and semilunar valve abnormalities (3.2%, 10/316). VSD accounted for the largest proportion (24.4%, 77/316) of the ventricular malformations. The total proportion of obstructive lesions in this group was much higher for the right than for the left side of the heart (18.4% (58/316) vs 9.5% (30/316), respectively). The spectrum of fetal CHD detected by echocardiography was much different compared to that accepted in the past. Complex defects were more common prenatally. Ventricular malformations were the largest constituent of all of the defects associated with fetal CHD, and VSD was the most common component of complex defects. Chinese fetal CHD encompassed more right-sided than left-sided obstructive lesions.

Indistinguishable tunable single photons emitted by spin-flip Raman transitions in InGaAs quantum dots.

This Letter reports all-optically tunable and highly indistinguishable single Raman photons from a driven single quantum dot spin. The frequency, linewidth, and lifetime of the Raman photons are tunable by varying the driving field power and detuning. Under continuous-wave excitation, subnatural linewidth single photons from off-resonant Raman scattering show an indistinguishability of 0.98(3). Under π pulse excitation, spin- and time-tagged Raman fluorescence photons show an almost vanishing multiphoton emission probability of 0.01(2) and a two-photon quantum interference visibility of 0.95(3). Lastly, Hong-Ou-Mandel interference is demonstrated between two single photons emitted from remote, independent quantum dots with an unprecedented visibility of 0.87(4).

Oleanolic acid inhibits hypertrophic scarring in the rabbit ear model.

BACKGROUND: Hypertrophic scarring, a common proliferative disorder of dermal fibroblasts, results from an overproduction of collagen and excessive deposition of extracellular matrix. Although treatment with surgical excision or steroid hormones can modify the symptoms, numerous treatment-related complications have been described. AIM: To investigate the effects of oleanolic acid (OA), a naturally occurring triterpenoid, on hypertrophic scarring in a rabbit ear model. METHODS: A rabbit ear model of hypertrophic scarring was used, with wounds produced with a biopsy punch. Oleanolic acid (2.5%, 5% and 10%) was applied once daily to the scars for 22 days. On postoperative day 28, the scars were excised, and the tissue used for histological examination and assays of the levels of collagens I and III, matrix metalloproteinase (MMP)-1 and transforming growth factor (TGF)-ß(1). The scar elevation index (SEI) was also determined. RESULTS: Treatment with different concentrations of oleanolic acid (OA) for 22 days significantly inhibited hypertrophic scarring in rabbit ear tissue. Levels of TGF-ß(1), collagen I and collagen III were significantly decreased and levels of MMP-1 significantly increased in the scar tissue. SEI was also significantly reduced. Histological findings showed significant amelioration of the scar tissue. CONCLUSIONS: OA suppresses hypertrophic scarring in the rabbit ear model and may be an effective cure for human hypertrophic scarring.

Catechin stimulates osteogenesis by enhancing PP2A activity in human mesenchymal stem cells.

SUMMARY: Using human mesenchymal stem cells, we identified catechin from a panel of herbal ingredients and Chinese traditional compounds with the strongest osteogenic effects. Catechin increased alkaline phosphatase activity, calcium deposition, and mRNA expression of Runx2 and osteocalcin. We further clarified the signaling pathway that catechin mediated to stimulate osteogenesis. INTRODUCTION: Human mesenchymal stem cells (hMSCs), useful as a species specific cell culture system for studying cell lineage differentiation, were examined as a tool to identify novel herbal ingredients and Chinese traditional compounds for enhancing osteogenesis. METHODS: Immortalized and primary hMSCs were induced in osteogenic induction medium in the presence of a variety of herbal ingredients and Chinese traditional compounds and osteogenic differentiation was evaluated by histochemical assays and quantitative RT-PCR. RESULTS: Using immortalized hMSCs, we first identified catechin, 18ß-glycyrrhetinic acid, baishao, and danggui with osteogenic properties, which enhanced calcium deposition at the dose without significant cytotoxic effects. Primary hMSCs were then applied for confirming the osteogenic effects of catechin, which increased alkaline phosphatase activity, calcium deposition, and mRNA expression of Runx2 and osteocalcin. We further found the extracellular signal-regulated kinase (ERK) pathway was downregulated upon stimulation with catechin. Catechin increased the level and activity of protein phosphatases 2A (PP2A) that dephosphorylates ERK kinase (MEK) and ERK. Further, PP2A inhibitor, okadaic acid, abolished the effect of catechin-mediated inactivation of ERK and stimulation of osteogenesis. The blocking effect of okadaic acid on osteogenesis was further reversed by PD98059, a specific inhibitor of MEK. Co-immunoprecipitation revealed the association of PP2A to both MEK and ERK. CONCLUSIONS: These studies propose catechin enhanced osteogenesis by increasing the PP2A level that inhibits the MEK and ERK signaling in hMSCs. These results prove the concept of using hMSCs as a convenient tool for rapid and consistent screening of the osteogenic herbal ingredients and traditional Chinese compounds.

Comparison between cells and cancer stem-like cells isolated from glioblastoma and astrocytoma on expression of anti-apoptotic and multidrug resistance-associated protein genes.

This study is to explore and compare the features of the cells and cancer stem-like cells (CSCs) isolated from both glioblastoma and astrocytoma on expression of anti-apoptotic and multidrug resistance-associated protein (MRP) genes. As a result, the mRNA expression of livin, livinalpha and MRP1 was up-regulated in human CSCs from 2 times to 85 times, but the gene expression of MRP3 was down-regulated from 0.09 times to 0.5 times. After just differentiation the mRNA expression of livin, livinalpha and MRP3 was up-regulated from 9 times to 64 times, but the mRNA expression of MRP1 was down-regulated from 0.01 times to 0.03 times. It is a rare report that glioma stem-like cells can be induced successfully from a grade 2-3 astrocytoma tissue. The properties of glioblastoma and astrocytoma stem-like cells on anti-apoptotic and MRP genes are: anti-apoptotic gene livin and survivin are elevated in CSCs but are the most increased in just differentiated CSCs; MRP1 gene is significantly increased and MRP3 is decreased in CSCs, but when differentiating the MRP3 gene starts a remarkable increase in CSCs; the expression of anti-apoptotic and MRP genes shows no differences between the CSCs isolated from glioblastoma and astrocytoma tissues.

Transplantation of microencapsulated genetically modified xenogeneic cells augments angiogenesis and improves heart function.

Cell-based gene therapy offers an alternative strategy for therapeutic angiogenesis for the management of myocardial infarction (MI). However, immune rejection poses a significant obstacle to the implantation of genetically engineered allogeneic or xenogeneic cells. In the present study, an ex vivo gene therapy approach utilizing cell microencapsulation was employed to deliver vascular endothelial growth factor (VEGF) to ischemic myocardium. Chinese hamster ovary (CHO) cells were genetically modified to secrete VEGF and enveloped into semipermeable microcapsules. In vitro assay indicated that the microencapsulated engineered CHO cells could secrete VEGF as high as 3852 pg ml(-1) per 48 h at day 8 after encapsulation. Then the microencapsulated CHO cells were implanted into the injured myocardium in a rat MI model, while engineered CHO cells, blank microcapsules and serum-free culture media were implanted as controls. The humoral immunity to xenogeneic CHO cells were evaluated and we found that the titer of anti-CHO antibodies was significantly lower in the microencapsulated CHO transplantation group than the group receiving unencapsulated CHO cells at two weeks after implantation. However, 1 week later, there was almost no difference between these groups. Histology and western blotting confirmed that the microencapsulated CHO cells maintained their original structure and VEGF secretion three weeks after implantation. The capillary density in the treatment region was also significantly higher in the microencapsulated CHO cell group than control groups, which was consistent with gross heart functional improvement. These data suggest that microencapsulated xenogeneic cell-based gene therapy might be a novel approach for therapeutic angiogenesis in ischemic heart disease.

Proteomic analysis of low-abundant integral plasma membrane proteins based on gels.

To characterize low-copy integral membrane proteins and offer some methods for human liver proteome projects, we fractionated highly purified rat liver plasma membrane (PM). PM was purified through two sucrose density gradient centrifugations, and treated with 0.1 M Na(2)CO(3), chloroform/methanol and Triton X-100. Proteins were separated by electrophoresis and submitted to mass spectrometry analysis. Four hundred and fifty-seven non-redundant membrane proteins were identified, of which 23% (105) were integral membrane proteins with one or more transmembrane domains. One hundred and fifty-three (33.5%) had no location annotation and 68 were unknown-function proteins. The proteins from different fractions were complementory. A database search for all identified proteins revealed that 53 proteins were involved in the cell communication pathway. More interestingly, more than 50% of the proteins had a protein abundance index concentration of less than 0.1 mol/l, and 12% proteins a concentration 100 times less than that of arginase 1 and actin.