High mobility group A2 protein (HMGA2), an architectural factor, is highly expressed in various cancer types including lung cancers. It is a candidate target for cancer therapy. RNAi is an effective gene silencing method with low cost and less time-consuming. It is possible to exploit this technology in therapy. Here, 5 siRNAs targeting Hmga2 gene (HMGA2 siRNA1-5) were designed and synthesized. MTT assay, colony formation assay, transwell assay and flow cytometry were used to evaluate the effects of these siRNAs on lung cancer cell lines (NCI-H446 and A549). Results from cell proliferation, clone formation, migration and apoptosis showed that HMGA2 siRNA1, 3, 5 could affect these aspects for both lung cancer cell lines. Among the five siRNAs, HMGA2 siRNA5 showed the greatest inhibition effects. The inhibition effects of HMGA2 siRNA5 are sequence specific and are not due to the induction of interferon response. Taken together, siRNAs targeting Hmga2 gene are potential candidates for lung cancer gene therapy.
Apoptosis ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colony-Forming Units Assay ; Gene Silencing ; Genetic Therapy ; HMGA2 Protein ; genetics ; metabolism ; Humans ; Interferons ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Point Mutation ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

The aim of this study was to evaluate the homing capabilities of hematopoietic stem/progenitor cells (HSPCs) derived from human placenta tissues (PT). Single cell suspension of human PT was prepared by mechanical method. The expression levels of homing-related adhesion molecules (HRAM) including CD11a, CD49d, CD44, CD49e, CD62L and CD54 on CD34(+) cells and the percentages of CD34(+) cells and their subpopulations in nucleated cells (NC) from fresh human PT, umbilical cord arterial blood (UCAB) and umbilical cord venous blood (UCVB) were detected by using flow cytometry. The results showed that the percentage of CD34(+) cells and CD34(+)CD38(-) cells in placenta were higher than those in UCAB and UCVB. There were no significant difference in percentage of HSPC between UCAB and UCVB. Placenta-derived CD34(+) cells strongly expressed CD11a, CD49d, CD44, CD49e and CD54, among which expression levels of CD49e and CD54 on placenta-derived CD34(+) cells were significantly higher than those on UCAB and UCVB-derived CD34(+) cells. While the percentage of CD34(+)CD62L(+) cells in placenta was only lower than that in UCVB. It is concluded that human placenta is rich in HSPC. Moreover, the expression levels of most HRAM in CD34(+) cells from PT are higher than those from UCAB and UCVB or are close to them. It suggested that HSPCs derived from PT might have stronger homing capabilities than those from UCB.
Antigens, CD34 ; biosynthesis ; Cell Adhesion Molecules ; biosynthesis ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; metabolism ; Humans ; Hyaluronan Receptors ; biosynthesis ; Integrin alpha5 ; biosynthesis ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Placenta ; cytology

The response rules of pressing pain on the back section in the Governor Vessel in patients with gastro-esophageal reflux disease (GERD) were studied to provide references for the diagnosis and treatment of GERD. Seventy-six cases of GERD were included into an observation group while 30 healthy volunteers were recruited into a control group. A mechanical measurement device of pressing pain that could measure the pain threshold was adapted to observe the pressing pain on the back section in the Governor Vessel in GERD patients and healthy volunteers. The test area is from spinous process of the 1st thoracic vertebra to that of the 12th thoracic vertebra (T1 -T12), including acupoints and non-acupoints on the Governor Vessel. As a result, in the observation group the pain threshold of T5-T7 spinous process clearance, which was the location of Shendao (GV 11), Lingtai (GV 10) and Zhiyang (GV 9), was lower than that in the control group (all P < 0.05). This result indicated that there was significant pressing pain in T5-T7 spinous process clearance in patients with GERD, which could be taken as an important auxiliary diagnosis and a new thinking method in the treatment of GERD with acupuncture.
Adolescent ; Adult ; Aged ; Diagnosis, Differential ; Female ; Gastroesophageal Reflux ; diagnosis ; physiopathology ; Humans ; Male ; Meridians ; Middle Aged ; Pressure ; Sensation ; Thoracic Vertebrae ; physiopathology ; Young Adult

Objective To explore the normal range of the insula volume of Chinese adults of the Han nationality and its relationship with age, which provide morphological data for the construction of database for Chinese Standard Brain.Methods This is a clinical multi-center study.One thousand Chinese healthy volunteers (age range = 18 to 70) recruited from 16 hospitals were divided into 5 groups, i.e.,Group A ( age range = 18 to 30), B ( age range = 31 to 40 ), C ( age range = 41 to 50 ), D ( age range =51 to 60), and E (age range =61 to 70).Each group contained 100 males and 100 females.All of the volunteers were scanned by MR using T1 weighted three-dimensional magnetization prepared rapid acquisition gradient echo sequence.After three dimension data reconstruction, the volumes of bilateral insula were manually measured.The volume of bilateral insula were compared by paired sample t test.The insula volumes were compared between male and female by independent sample t test, and the differences among 5 age groups were compared by one-way ANNOVA.The relationship between the volumes of insula and age,sex or cerebral volume were analyzed using bivariate correlation, respectively.Results The left snd right side volume of insula before standarized were (7764±1165) and (7387±1128) mm3 respectively, after standarized were (8413±1201 ) and (7871±1140) mm3 respectively.The left insula volume were significant larger than that of right(t = - 10.565, - 16.014,P <0.01 ).The left and fight volume of insula were (8146±1181 ) and (7735±1113) mm3 for male, and (7393±1022) mm3 and (7050±1038) mm3for female.The left and right insula volumes for male were larger than the female's(t = 10.934,9.945 ,P <0.01 ).The left and right insula volume of male after standarized were (8779±1230 ) and (8224±1081 )mm3, female were (8043±1054) and (7515±1091 ) mm3 ,the left and right insula volume of male were larger than the female's ( t = 4.858,4.632, P < 0.01 ).The left insula volumes among Group A, B, C, D,E before standarized were ( 8268±1221 ), ( 8067±1107 ), ( 7869±1109), ( 7603±1111 ), ( 6997±934 )mm3 respectively,the right were (8028±1156), (7636±1075), (7294±986), (7249±1068), (6717±916) mm3 respectively, there were significant differences among 5 groups between left and right insula volume(F= -0.361,-0.337,P <0.01 ).The left insula volume of A,B,C,D,E after standarized were ( 9093±1105 ), ( 8679±965 ), ( 8810±1136), ( 8202±980), ( 7273±940 )mm3, the right were ( 8694±1005), (8136±1100), (8034±910), (7496±990), (6989±932) mm3, there were significant differences among 5 groups between left and right insula volume(F = -0.490, -0.512,P < 0.01 ).There was significant negative correlation between the volume and age ( before standarizded:r = -0.361, -0.337, after standardized r = -0.490, -0.512, P<0.05).Before and after standardized, there was significant correlation between the volume of right and left insula and cerebral volume ( r = 0.470,0.459, P < 0.05 ).Before and after standardized, there was significant correlation between the volume of right and left insula and weight( r = 0.141, 0.092, P < 0.05 ).Conclusions 1.5 T MR scanner has high resolution, for distinguishing the white matter from the gray matter, and provide morphological data of insula for the construction of database for Chinese Standard Brain.

Placentation,which is critical for maternal-fetal exchange of nutrients and gases,is a complicated process comprising stepwise vasculogenesis and angiogenesis.Hypoxia caused by impairedtrophoblast invasion may cause various angiogenic abnormalities in human placenta.The Notchl signaling pathway plays an important role in the regulation of angiogenesis.The angiogenesis of human umbilical vein endothelial cells (HUVECs) under normal/hypoxic conditions and the mRNA/protein level of Notchl/Dell4/Jaggedl were investigated in this study.The effects of DAPT/JAG-1 on the migration of HUVECs were also assessed by cell wound healing assay,so as to discover the possible role of notchl signaling pathway in the angiogenesis of human placenta.The results showed that angiogenic ability of HUVECs was seriously reduced under hypoxic conditions.The mRNA and protein levels of Notchl/Dell4/Jaggedl were decreased in the hypoxic group compared to the control one.In addition,the migration capability of HUVECs was significantly obstructed when treated with DAPT and under hopoxic condition,but promoted when treated with JAG-1.The above results demonstrate that hypoxia downregulates the angiogenesis in human placenta via Notch 1 signaling pathway.

A large amount of evidence has showed that sexually transmitted infection is an important synergistic factor of human immunodeficiency virus (HIV) infection.Therefore, this paper reviews the current situation of sexually-transmitted diseases (STD) and HIV infection, introduces HIV prevention and intervention measures and problems for STD patients at home and abroad, and proposes that behavior-psychology-society integrated intervention model should be constructed based on the characteristics of STD patients.

Objective To explore the possible pathogenesis of chronic posthemorrhagic hydrocephalus by observing the expression of AQP1 on rats after experimental intraventricular hemorrhage. Methods Fourty-five rats were randomly divided into normal control(n=5),sham-operated(n=20)and experimental(n=20)groups.The 0.1mL saline and 0.1mL citrated autologous blood were injected into the lateral ventricle of the rats in the sham-operated and the experimental groups,respectively.The later two groups were divided into four subgroups according to different time points at 3,7,14 and 30 d(n=5).The change of protein expression of AQP1 at different time points of bleeding were detected by immunohistochemical techniques and that of mRNA expression of AQP1 was obsevred by in situ hybridization,respectively.and then,the possible pathogenesis of chronic posthemorrhagic hydrocephalus was discussed. Results At the time point of 30 d after intraventricular hemordmge,chronic hydrocephalus appeared in 4 rats(80%)in the experimental group.High protein expression of AQP1 was found in the apical of cuboidal epithelium of choroids plexus,the ependyma, the pia mater, the arachnoid and the dura in the normal control group;the protein expression of AQP1 gradually weakened in the experimental group 3 d aftea intraventricular hemorrhage and dropped to the bottom on the 14th d,which was significantly different from the normal control and sham-operative groups(P<0.05).The mRNA expression of AQP1 was weaker than the protein expression of AQP1,and the expression locations of them were basically in concordance. Conclusion AQP1 is involved not only in the secretion of cerebrospinal fluid,but also in the process of the CSF absorption.The decrease of CSF absorption induced by the decreased expression of AQP1 after intraventrieular hemorrhage in rats may also relate to the development of chronic posthemorrhagic hydrocephalus.

OBJECTIVETo explore the acute effects of testosterone at the physiological level on PGF2alpha-induced increase in intracellular Ca2+ in cultured vascular smooth muscle cells (VSMCs).

METHODSVSMCs from the thoracic aorta of male Sprague-Dawley rats were cultured using the explant method. The subconfluent VSMCs were incubated with serum-free medium for 24 hours to obtain quiescent non-dividing cells and then treated with the indicated agents. For the measurement of [Ca2+]i, the VSMCs were loaded with fura-2. Changes of [Ca2+]i were determined ratiometrically with a Nikon TE-2000E system.

RESULTSThe resting level of [Ca2+]i was around 100 nmol/L in the VSMCs. Exposing cells to perfusate containing 10 micromol/L PGF2alpha triggered an immediate and transient peak in [Ca2+]i, which gradually decreased afterwards. Interference at the peak with the physiological concentration (40 nmol/L) of testosterone significantly decreased the peak-to-baseline time of [Ca2+]i, compared with ethanol vehicle (104.9 +/- 27.0 s vs 153.5 +/- 40.4 s, P < 0.01). Pretreatment with testosterone at 40 nmol/L for 2 minutes also reduced the peak-to-baseline time of [Ca2+]i significantly in comparison with the ethanol control (120.6 +/- 32.0 s vs 151.4 +/- 27.4 s, P < 0.01), but it had no significant effect on the peak level of PGF2alpha-induced intracellular Ca2+ (390.0 +/- 126.0 nmol/L vs 403.4 +/- 160.7 nmol/L, P > 0.05).

CONCLUSIONTestosterone at physiological concentration inhibits PGF2alpha-induced Ca2+ fluxes, probably via receptor-operated calcium channels by a non-genomic mechanism in VSMCs, which may be involved in the vasodilatory effect of testosterone.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Dinoprost ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; metabolism ; Rats ; Rats, Sprague-Dawley ; Testosterone ; metabolism ; physiology

OBJECTIVETo determine the presence of membrane testosterone receptors in cultured vascular smooth muscle cells (VSMC), and investigate their relationship with classical intracellular androgen receptors (iAR).

METHODSVSMCs were cultured from the thoracic aorta of male Sprague-Dawley rats by the explant method. Subconfluent VSMCs were incubated with serum-free medium for 24 h to obtain quiescent non-dividing cells, and then treated with the indicated agents. The aliquots of VSMCs were labeled with testosterone-BSA-FITC (T-BSA-FITC) and analyzed by flow cytometry. Classical iARs in intact- and permeabilized-cells were detected with anti-iAR antibodies and FITC-labeled secondary antibodies by immunofluorescence, followed by flow cytometry analysis.

RESULTSIncubation of VSMCs with T-BSA-FITC obviously increased their relative fluorescence intensity at 10 sec as compared with the untreated controls (P < 0.01), and so did it at 10 min in comparison with the treatment with BSA-FITC alone or together with free testosterone (P < 0.01). Pretreatment with iAR antagonist flutamide exhibited no significant influence on the relative fluorescence intensity of VSMCs (P = 0.318). Traditional iARs were not detectable on the surface of intact VSMCs, although permeabilized cells contained iARs.

CONCLUSIONVSMCs contain testosterone receptors in the plasma membrane, and these membrane receptors are not identical to classical iARs.

Animals ; Cells, Cultured ; Male ; Membrane Proteins ; metabolism ; Muscle, Smooth, Vascular ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; metabolism ; Testosterone ; metabolism

OBJECTIVETo improve the understanding of progressive transformation of lymph node germinal centers (PTGC) and to explore its clinical, histopathologic and immunohistochemical features and the differential diagnosis between the related disease of germinal center hyperplasia.

METHODSThe clinical manifestation, laboratory bindings, treatment and outcome of a patient with PTGC were presented.

RESULTSThe main manifestation of the patient was painless peripheral lymphadenopathy. Histopathologic examination of an axillary lymph node showed reactive follicular hyperplasia and the progressive transformation changes germinal centers. The borderline between the germinal center and the mantle layer was obscured. The cells in the progressive transforming germinal centers were positive for CD20(+), CD5(+), CDw75(+).

CONCLUSIONPTGC is a rare lymphoid disorder. Histopathology and immunohistochemistry are important basis of the diagnosis.

Diagnosis, Differential ; Germinal Center ; Humans ; Hyperplasia ; Lymph Nodes ; Lymphatic Diseases