Matrine inhibits proliferation and promotes autophagy and apoptosis in non-small cell lung cancer cells by deactivating PI3K/AKT/mTOR pathway / 南方医科大学学报

OBJECTIVE@#To investigate the inhibitory effect of matrine on the proliferation of human non-small cell lung cancer (NSCLC) and explore the possible molecular mechanism.@*METHODS@#Cultured human NSCLC A549 cells were treated with 0.4, 0.8, 1.2, 1.6, and 2.0 g/L matrine for 24, 48 or 72 h. CCK-8 assay was used for measuring the changes in A549 cell viability. The morphological changes of the cells were observed under a fluorescence microscope, and flow cytometry was employed for analyzing the cell apoptosis. The effects of matrine and the PI3K specific inhibitor LY294002 (10 nmol/L) on AKT pathway and autophagy-related proteins in A549 cells were investigated using Western blotting.@*RESULTS@#Matrine significantly inhibited the proliferation of A549 cells in a time- and dose-dependent manner ( < 0.05). At the concentration of 1.6 g/L or higher, matrine caused obvious cell shrinkage and fragmentation and significantly increased floating cells; autophagy vacuoles could be observed in the cells after acridine orange staining. Within the concentrations range of 0.8-1.6 g/L, matrine time- and dosedependently increased the cell apoptosis. Treatment of the cells with 1.6 g/L matrine and 10 nmol/L LY294002 resulted in significantly lowered expressions of p-AKT and p-mTOR proteins and increased the expression of light chain 3B (LC 3B), an autophagy-related protein, as compared with those in the control cells ( < 0.05).@*CONCLUSIONS@#We demonstrate that matrine inhibits the proliferation and induces autophagy and apoptosis of A549 cells by deactivating AKT pathway, suggesting the potential of matrine as an anti-cancer agent for lung cancer.

Treatment of experimental hepatic fibrosis in rats by transplanting with mesenchymal stem cells derived from human umbilical cord and Interleukin-10 / 中国免疫学杂志

Objective:To observe the conditions of isolation,purification and culture of mesenchymal stem cells(MSCs) in vitro derived from human umbilical cord,and to observe the repairing effect on experimental hepatic fibrosis in rats by transplantation with MSCs only or MSCs with IL-10.Methods:The MSCs derived from human umbilical cord were isolated,purified and cultivated in vitro.The model of hepatic fibrosis induced by CCl4 was established.After the model of hepatic fibrosis was succeeded,the rats were divided into three groups:model group,MSCs transplantation group,MSCs transplantation with IL-10 group.MSCs transplantation group was given 1×106 MSCs by the tail vein injection at the first 1 weeks,2 weeks and 3 weeks.MSCs transplantation with IL-10 group not only was given MSCs the same with MSCs transplantation group, but also was injected of IL-10 ( 4 μg/kg ) in abdomen, 4 times a week.After 4 weeks,the rats were executed,rats blood was collected to observe the changes of liver function and the index of liver fibro-sis.The liver HE staining was carried out to observe the changes of liver pathology.Expression of MMP-2 mRNA was detected by RT-PCR method,expression of TGF-βprotein was detected by Western blot method.Results:The rat liver function and the index of liver fibrosis improved obviously with a significant difference between model group , MSCs transplantation group and MSCs transplantation with IL-10 group after transplanting,MSCs with IL-10 was better than MSCs only.Liver tissue of rats with hematoxylin eosin staining suggested,liver fibrosis was obviously improved,and the MSCs transplantation with IL-10 group was best.MMP-2 mRNA expression was significantly higher between normal group and model group,MSCs transplantation group with a significant difference (P0.05).TGF-βprotein expression of other groups was higher than normal group with a significant (P<0.05),and MSCs transplantation with IL-10 group was the largest decrease than normal group.Conclusion:MSCs only or MSCs with IL-10 transplantation can improve the biochemical characteristics of rat peripheral blood and liver histological structure,and with IL-10 wan better than MSCs only.MSCs and IL-10 play a role in the treatment of liver fibrosis through lowering the expression of MMP-2 and the expression of TGF-βprotein.

Pharmaceutical Care for an Elderly Patient with Non-small Cell Lung Cancer Complicated with Severe Bone Marrow Suppression / 中国药师

Objective:To explore the pharmaceutical care for the elderly cancer patients complicated with bone marrow suppres-sion by clinical pharmacists. Methods:Clinical pharmacists performed the pharmaceutical care through the evaluation of chemothera-py, symptomatic treatment for bone marrow suppression and infection prevention after bone marrow suppression etc. Results:The bone marrow suppression was alleviated without bleeding related with the decrease of platelet and infection. Conclusion:The pharmaceutical care performed by clinical pharmacist can enhance the safety and effectiveness of medication.

Effects of Panax notoginseng saponins on proliferation, apoptosis and cell cycle of K562 cells in vitro and the mechanisms / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of Panax notoginseng saponins (PNS) on the proliferation, apoptosis and cell cycle of K562 cells and explore the molecular mechanisms underlying these effects.</p><p><b>METHODS</b>PNS-induced growth inhibition of K562 cells was detected by MTT assay; the cell apoptosis was evaluated by AO/EB staining and Annexin V-FITC/ PI staining; flow cytometry was used to detect cell cycle changes in the treated cells. The mRNA expressions of the molecules in mTOR signaling pathway were examined by RT-PCR, and the cellular expressions of cleaved caspeas-3, cyclin D1 and major proteins in mTOR signaling pathway were detected using Western blotting.</p><p><b>RESULTS</b>MTT assay showed that treatment with 100-800 µg/mL PNS significantly inhibited the proliferation, promoted the cell apoptosis, and caused cell cycle arrest in G0/G1 phase in K562 cells. Western blotting revealed increased protein expression of cleaved caspase-3 and decreased expression of cyclin D1 in PNS-treated cells, in which the proteins expressions of mTOR, p-mTOR, p-p70S6K and p-4E-BP 1 and the mRNA expression of mTOR were all decreased.</p><p><b>CONCLUSION</b>PNS can inhibit the proliferation, induce apoptosis and cause cell cycle arrest in K562 cells possibly by up-regulating cleaved caspase 3 and down-regulating cyclin D1 and mTOR signaling pathway.</p>

Audiologic assessment of the children with cleft lip/palate / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the audiologic characteristics of the children with cleft lip and/or palate.@*METHOD@#Sixty-two children with cleft lip/palate were enrolled in the study. Tympanometry, DPOAE, ABR were tested in all the chidren.@*RESULT@#Regarding the ABR threshold as the diagnostic criteria, 51 (41.13%) ears had hearing loss, mainly moderate. Sixty-three (59.43%) ears had abnormal tympanometry, which meant the disorder function of the middle ear. Twenty-six (56.52%) ears didn't pass the DPOAE.@*CONCLUSION@#The proportion of the hearingloss in the children with cleft lip/palate was large. There was a good coherence among the three objective examinations when assess the children with cleft lip/palate.

Vein transplantation of human umbilical cord-derived mesenchymal stem cells for treatment of hepatic fibrosis in rats / 中国组织工程研究

BACKGROUND:Recent studies verified that other tissues-derived stem cells can be homing to the liver, possibly participate in the regeneration of liver tissues, which provides new hope for stem cells in treatment of liver disease. OBJECTIVE:To observe isolation and culture of human umbilical cord-derived mesenchymal stem cells, to observe the repairing effect of umbilical cord-derived mesenchymal stem celltransplantation on hepatic fibrosis, and then to provide a reliable theoretical basis for further clinical application of umbilical cord-derived mesenchymal stem cells. METHODS:Human umbilical cord-derived mesenchymal stem cells were isolated and purified by natural adherent method, and then cultured and amplified in vitro. A rat model of hepatic fibrosis was prepared using subcutaneous multi-point injection of CCl4. A total of 22 rat models were randomly assigned to model injury group (n=11) and celltransplantation group (n=11). At 1, 2 and 3 weeks after model induction, the rats in the celltransplantation group were treated with 1×106 umbilical cord-derived mesenchymal stem cells via caudal vein. Four weeks later, the rats were sacrificed. Blood of rats was col ected from each group to detect liver functions. The liver was removed to receive hematoxylin-eosin staining, and pathological changes were observed. The number and distribution of Kupffer’s cells were observed using immunohistochemistry. The localization of umbilical cord-derived mesenchymal stem cells from treatment group was observed using immunohistochemistry.RESULTS AND CONCLUSION:After umbilical cord-derived mesenchymal stem cells were infused into rats with cirrhosis via caudal vein, liver function was significantly improved, which showed significant differences as compared with the control group (P<0.05). Hematoxylin-eosin staining revealed that hepatic fibrosis was apparently improved. Immunohistochemistry results demonstrated that the number of Kupffer’s cells was obviously reduced, and BrdU-labeled umbilical cord-derived mesenchymal stem cells were visible in rat liver of the treatment group using anti-BrdU antibody. These results suggested that umbilical cord-derived mesenchymal stem celltransplantation could improve biochemical characteristics of rat peripheral blood and histological structure of liver.

Lentivirus-mediated stable Fas gene silencing in human umbilical cord-derived mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentivirus-mediated vector for RNA interference (RNAi) of Fas and establish a human umbilical cord-derived mesenchymal stem cells (UC-MSCs) line with stable Fas gene silencing.</p><p><b>METHODS</b>Four short hairpin RNA sequences targeting the coding region of human Fas mRNA were designed. The synthesized oligonucleotides were ligated with the lentivirus vectors harvested from BamHI and EcoRI double digestion of LV3 recombinant vector. The recombinant lentivirus vectors were transfected into the packaging cells 293T, and the lentivirus titers were determined. Cultured UC-MSCs were infected with the lentivirus, and real-time PCR and Western blotting were used to detect the expressions of Fas mRNA and protein in the transfected cells.</p><p><b>RESULTS</b>Restriction digestion and DNA sequencing showed that the lentiviral vectors were successfully constructed, and the titer of lentivirus reached 3 × 10⁸ TU/ml in the packaging cells. Real-time PCR and Western blot demonstrated significantly suppressed Fas gene expression in UC-MSCs after infection with the recombinant lentivirus.</p><p><b>CONCLUSION</b>Lentivirus-mediated RNAi can effectively inhibit Fas gene expression in cultured UC-MSCs.</p>

Efficacy and safety of sublingual immunotherapy with house dust mite extract in poly allergen sensitized children with allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To compare the efficacy of sublingual immunotherapy (SLIT) with standardized house dust mite extract in mono sensitized and polysensitized children with allergic rhinitis.@*METHOD@#One hundred and fifty-seven children who were sensitized to house dust mites and treated with SLIT for house dust mites for at least 1 year were studied. The monoallergen sensitized group included patients who were sensitized to Dermatophagoides pteronyssinus and/or Dermatophagoides farinae (n=92). The polyallergen sensitized group included patients who were simultaneously sensitized to house dust mites and other allergens (n = 65). A standardized extract of house dust mites was used for immunotherapy. Antiallergic medication and the total nasal symptom score (TNSS) were evaluated before and 1 year after SLIT.@*RESULT@#One hundred and twenty-five children completed 1-year SLIT. The TNSS improved significantly after SLIT in both groups, with monoallergen sensitized group 11.42 +/- 1.60 vs 3.55 +/- 1.57 (t=30.03, P0.05). The AMSs were decreased significantly after SLIT in both groups, with monoallergen sensitized group 1.62 +/- 0.44 vs 0.56 +/- 0.37 (t=15.01, P0.05).@*CONCLUSION@#In polysensitized allergic rhinitis patients, SLIT for D pteronyssinus and/or D farinae produced improvements in both nasal symptoms and rescue medication scores comparable to those in mono sensitized patients. SLIT for D pteronyssinus and/or D farinae should be considered in polysensitized allergic rhinitis patients.

The correlation between the status of circumferential resection margin and the prognosis of patients With middle and lower rectal carcinoma / 中华普通外科杂志

Objective To study the relationship between circumferential resection margin status and prognosis of patients with middle and lower rectal cancer.Methods Specimens from 49 patients with middle and lower rectal cancer undergoing total mesorectal excision were studied by the large slice pathologic technique.The local recurrence,metastasis and five-year survival rate were evaluated by Kaplan-Meier Survival analysis.The related clinicopathologic factors were also analyzed.Results The cancer involvement rate of the circumferential resection margins was 24%(12/49).The overall local recurrence rate was 12%(6/49),the distant metastasis and recurrence rate was 27%(13/49),and the five-year survival rate was 67%(33/49).For the 12 patients in which the eircumferential resection margin was tumor positive.the local recurrence rate was 33%compared with 5%in those with negative circumferential resection margin(X2=6.577,P=0.010),distant recurrence was 50%compared with 19%in those with tumor negative margin(X2=4.491,P=0.034).Kaplan-Meier survival analysis showed that patient's survival time was statistically correlative with the circumferential resection margin status(log-rank.P=0.009).Five-year survival rate was 33%in patients with positive circumferential resection margin,compared with 78%in those with negative margins.Tumor diameter(X2=4.451,P=0.035),T staging (X2=20.283,P=0.000),N staging(X2=7.773,P=0.018),the distance away from the anocutaneous line(X2=6.502,P=0.04),tumor location(X2=4.421,P=0.035)and operation type(X2=5.754,P=0.016)were significantly correlated with the circumferential resection margin status of the middle and lower rectal cancer.Conclusions The circumferential resection margin status was an important predictor of local and distant recurrence as well as survival of patients with middle and lower rectal carcinoma.and the status is significantly correlated with tumor diameter,T staging,N staging,the distance away from the anocutaneous line,tumor location and operation type.

Etiology study on hand, foot and mouth disease in children in Beijing during 2007 to 2008 / 中华检验医学杂志

Objective To investigate the etiological agents of hand, foot and mouth disease (HFMD) in children in spring and summer from 2007 to 2008 in Beijing and the characteristics of the disease by virus isolation and to provide the scientific evidence for prevention and treatment for HFMD. Methods During April to August, 2007 and May to September, 2008, 356 clinical specimens including 255 throat swabs and 101 vesicle fluids were collected from 256 patients with HFMD who visited the Children's Hospital Affiliated to Capital Institute of Pediatrics and children with severe HFMD with neural system complications from Ditan Hospital and Youan Hospital All of the specimens were inoculated into Vero cells for virus isolation. After the cell pathogenic effects (CPE) appeared, the isolates were identified by RT-PCR with the universal primers within 5'untranslated region of enterovirus and typed by specific primers for VP1 gene of EV71 and CA16, respectively. The throat swabs from all of 10 severe HFMD were tested for enterovirus by RT-PCR addition to virus isolation. Results Out of 256 patients, 188 were positive for enterovirus by virus isolation, with the overall positive rate of 73.4%. Among the 356 clinical specimens collected from these 256 patients, 239 enterovirus strains were isolated with the overall positive rate of 67.1%. The positive rate for virus isolation from vesicle fluid samples was 75.2% which was higher than the positive rate of isolation from throat swabs (63.9%), but the time for CPE appearing in cell culture showed no significant difference. The positive rate of virus isolation from throat swabs from children with severe HFMD was 50% (5/10) which was lower than overall positive rate (73.4%) from regular HFMD. The RT-PCR typing for virus isolates revealed that among 45 enterevirus strains isolated from the specimens collected in 2007 by the universal primer pairs, 43 were CAI6 (95.6%, 43/45) and 2 were EV71 (4.4%, 2/45), whereas for the specimens collected in 2008, out of 143 enterovirus isolates by PCR with universal primers, 117 were EV71 (82.4%, 117/142) and 24 were CA16 (16.8%, 24/142). All of 10 severe cases were positive for EV71 by RT-PCR directly from clinical specimens. Conclusion CA16 and EVT1 were the etiological pathogens of HFMD in Beijing during 2007 to 2008 HFMD seasons. The dominant type of enterovirus was different between 2007 and 2008. Enterovirus type CA16 was predominant in 2007, whereas EV71 was predominant in 2008. All of severe cases of HFMD in children in this study were caused by EV71.

HPLC Content Determination of Luteolin in Callicarpa nudiflora Hook.et Arn / 中药新药与临床药理

Objective To determine the content of luteolin in Callicarpa nudiflora Hook.et Arn..Methods HPLC was performed with chromatographic conditions as follows:Alltima C18 column(250 mm? 4.6 mm,5 ? m)with column temperature at 25 ℃,the mobile phase of methanol-water-phosphate(55:45:0.4),flow rate at 1 mL/min,and the detection wavelength at 348 nm.Results The linearity of luteolin was good in the range of 0.032 46～ 0.129 84 ? g(r=0.999 5).The average recovery was 99.52 % and RSD was 1.58 %.Conclusion The method is simple,feasible and reproducible,and can be used to control the quality of Callicarpa nudiflora Hook.et Arn..

RP-HPLC Determination of Paeoniflorin in Shiquan Dabu Granules / 中药新药与临床药理

Objective To establish a content determination method of paeoniflorin in Shiquan Dabu Granules.Methods A RP-HPLC method was adopted.A Hypersil BDS C18(4.6 mm?200 mm,5 ?m)column served as the stationary phase,methanol-water(15:45) as mobile phase,and the detection wavelength was 230 nm.Results Paeoniflorin had a good linearity in the range of 0.104～0.519 ?g(r=0.999 2).The recovery was 98.77 %and RSD was 1.77 %.Conclusion The method is simple,accurate,reproducible,and can be used to control the quality of Shiquan Dabu Granules.