OBJECTIVETo observe p21-activated kinase 6 (PAK6) expression and its possible role after spinal cord injury (SCI) in adult rat.

METHODSSprague-Dawley rats were subjected to spinal cord injury. To explore the pathological and physiological significance of PAK6, the expression patterns and distribution of PAK6 were observed by Western blot, immunohistochemistry and immunofluorescence.

RESULTSWestern blot analysis showed PAK6 protein level was significantly up-regulated on day 2 and day 4, then reduced and had no up-regulation till day 14. Immunohistochemistry analysis showed that the expression of PAK6 was significantly increased on day 4 compared with the control group. Besides, double immunofluorescence staining showed PAK6 was primarily expressed in the neurons and astrocytes in the control group. While after injury, the expression of PAK6 was increased significantly in the astrocytes and neurons, and the astrocytes were largely proliferated. We also examined the expression of proliferating cell nuclear antigen (PCNA) and found its change was correlated with the expression of PAK6. Importantly, double immunofluorescence staining revealed that cell proliferation evaluated by PCNA appeared in many PAK6-expressing cells on day 4 after injury.

CONCLUSIONThe up-regulation of PAK6 in the injured spinal cord may be associated with glial proliferation.

Animals ; Astrocytes ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; metabolism ; Up-Regulation ; p21-Activated Kinases

OBJECTIVETo investigate the expression and relationship of p27(kip1) and its nuclear export factor Jab1 during proliferation process of lymphoma cell.

METHODSJurkat and Raji cells were treated with serum starvation and then serum release. The protein and mRNA expression of p27(kip1), Jab1 in the cells were detected by Western blot and RT-PCR respectively. LMB were used for stimulating Jurkat cells during their proliferation process, and then the expression changes of p27(kip1) and Jab1 were detected. An eukaryotic expression plasmid(pcDNA3. 1-myc) containing Jab1 was constructed. Jurkat cell were transfected in vitro with or without pcDNA3. 1-myc-Jab1. Double immunolabelling was used to identify the localization of p27(kip1). Immunoprecipitation was used to detect the combination of p27(kip1) and Jab1.

RESULTSThe growth of Jurkat and Raji cells were blocked by serum starvation. The total protein amount of p27(kip1) increased while that of Jab1 decreased. The reverse changes were happened after serum release, but the mRNA expression of p27(kip1) has no significant change. LMB could inhibit the cell proliferation caused by serum release. The expression of p27(kip1) was up-regulated and Jab1 down-regulated when Jurkat cells were treated with LMB. After pcDNA3. 1-myc-Jab1 infected Jurkat cells for 48 h, the distribution of p27(kip1) was translocated from nucleus into cytoplasma. p27(kip1) and Jab1 could form compound in Jurkat and Raji cells detected by Immunoprecipitation.

CONCLUSIONJab1 may influence the location and expression of p27(kip1) through integrating with p27(kip1), and then participates in regulating the growth of NHL cell through interfering with the function of p27(kip1).

COP9 Signalosome Complex ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Jurkat Cells ; Peptide Hydrolases ; metabolism ; Plasmids ; RNA, Messenger ; metabolism ; Transfection

OBJECTIVETo investigate the method for creation of auriculocephalic sulcus.

METHODSThe reconstruction was performed 4-12 months after the first surgery. Skin incision was made 5mm posterior to the outer margin of the auricle. The ear framework was elevated with a thick fascia at the deep surface. The costal cartilage banked at the first operation was shaved and transplanted to the deep surface of the concha with sutures. The position and angle of the ear framework was adjusted to be familiar to the healthy ear. The auriculocephalic angle was slightly larger than that in the contralateral ear. Two flaps were designed at the upper and lower area of reconstructed ear and rotated to cover the cartilage. The wound at the donor site was closed with skin graft.

RESULTSA total of 72 patients were treated. All the flaps survived completely. 51 patients were followed up for 3-24 months with satisfactory results. The auriculocephalic sulcus maintained at about 20-30 degree.

CONCLUSIONSIt is a simple, safe and reliable method to create a auriculocephalic sulcus with two random skin flaps from mastoid area combined with skin graft.

Adolescent ; Cartilage ; transplantation ; Dermatologic Surgical Procedures ; methods ; Ear ; Ear Auricle ; surgery ; Ear Deformities, Acquired ; surgery ; Fascia ; Humans ; Mastoid ; Ribs ; Skin Transplantation ; methods ; Surgical Flaps ; transplantation

OBJECTIVETo explore the mechanism of acupuncture in treatment of spinal cord injury (SCI).

METHODSAdult Wistar rats were used to make SCI model by Allen's method. The SCI rats were treated with electroacupuncture (EA) at acupoints of the Governor Vessel for 3 days, 1 week, 2 weeks or 4 weeks. Normal group and spinal cord injury group were used as controls. The number and morphology of astrocytes in each group were investigated by electron microscope, immunohistochemistry and in situs hybridization methods. The expression of glial fibroblast acid protein (GFAP) mRNA in the injured spinal cord was detected by reverse transcription polymerase (RP-PCR).

RESULTSThe mitochondria and ribosomes of astrocytes in the EA group increased. The number of astrocytes increased after SCI, in gray matter being more than that in the white matter, in the caudal being more than that in the rostral. The expression of GFAP mRNA in the EA group was significantly lower than that in the control group.

CONCLUSIONElectroacupuncture can inhibit the reactive proliferation of the astrocytes after spinal cord injury and prevent formation of the glial scar.

Animals ; Astrocytes ; Electroacupuncture ; Humans ; Rats, Sprague-Dawley ; Rats, Wistar ; Spinal Cord Injuries ; therapy

OBJECTIVETo investigate the epidemiological characteristics of human papillomavirus (HPV) infection in men attending a sexually transmitted diseases (STD) clinic in Hangzhou area.

METHODSMale subjects (n=375) aged 18-70 years, attending the STD clinic were recruited. Urethral swabs were assessed for HPV DNA using polymerase chain reaction (PCR) with the consensus primers MY09/11. HPV genotypes of positive PCR products were determined by restriction fragment length polymorphisms and direct sequence analysis.

RESULTSOf the 375 swabs collected, 305 (81.3%) yielded sufficient DNA for the subsequent HPV analysis. Among the 305 subjects, the prevalence of HPV was 13.8%. Nononcogenic HPV types were found in 8.5% (26/305) of subjects, oncogenic types in 4.3% (13/305), and multiple types in 1.0% (3/305). The prevalence of HPV infection was higher in subjects from urban area than in those from rural area (P < 0.05). The prevalence was also higher in those who received fewer years of education (P < 0.05) and those who had more sex partners (P < 0.05).

CONCLUSIONSHPV infection among men at high risk is not uncommon. The detection rate of HPV DNA is significantly related to some sociodemographic factors, such as residence, educational level and the number of sex partners.

Adolescent ; Adult ; Aged ; Ambulatory Care Facilities ; China ; epidemiology ; Humans ; Male ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; epidemiology ; virology ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sexually Transmitted Diseases ; prevention & control

OBJECTIVETo characterize the deficiency of the mRNA expression of specific protein (SP3) gene in peripheral blood mononuclear cells (PBMCs) from Chinese patients with multiple sclerosis (MS) and study its correlation with the disease phenotypes.

METHODSFifty-six patients with definite MS were collected and total RNA was extracted from their PBMCs. Specific primers corresponding to SP3 gene were designed and the mRNA expression of SP3 gene was detected by reverse transcriptase-PCR (RT-PCR) method. The deficiency of SP3 expression was compared among MS patients, irrelevant disease group and normal controls.

RESULTSOf the 56 MS cases, 23 (41.1%) were SP3-deficient. In contrast, the frequency of SP3-deficiency in normal subjects and irrelevant disease controls was 8.6% (5/35) and 14.3% (4/27), respectively. The frequency of the SP3-expression deficiency in MS patients was significantly higher than that in both control groups (P< 0.01). Within the MS cases, the scores of expanded disability status scale (EDSS) in the SP3-expressing subjects were significantly different from that in the SP3-deficient ones in the stable, but not in the active, phase of MS (P< 0.05).

CONCLUSIONAuthor's observation suggested that deficient expression of SP3 gene occurs in Chinese MS patients, and that the SP3 expression may correlate with the clinical manifestations of MS and play roles in its immunological pathogenesis.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Multiple Sclerosis ; genetics ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sp3 Transcription Factor ; genetics ; Young Adult

OBJECTIVETo study the effect of all-trans retinoic acid (ATRA) on U937 cell growth and its mechanism.

METHODSCell cycle was detected by flow cytometry (FCM), expressions of cell cycle associated protein and the p27 related protein were detected by Western blot. The binding of P27 and Skp2 was detected by immunoprecipitation.

RESULTSFCM displayed that ATRA could inhibit the proliferation of U937 cells. At 72 h on 1 micromol/L ATRA treatment, 72% of the cells were arrested at G0/G1 phase. Western blot displayed that ATRA could decrease the expression of cyclin A, up-regulate the expression of p21 and p27, and down-regulate the expression of p27 related proteins Skp2. p27 could bind with Skp2 in U937 cells as detected by immunoprecipitation.

CONCLUSIONATRA may arrest the proliferation of U937 cells through the reduction of Skp2 expression, and finally the induction of the accumulation of p27.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; S-Phase Kinase-Associated Proteins ; metabolism ; Tretinoin ; pharmacology ; U937 Cells

Objective To investigate the protective effect of modified Qianjin Weijing Decoction(MQJWJD)on TNF-α and NF-κB in rats with lung injury induced by particulate matter; To discuss relevant mechanism of action. Methods A tracheal drip 15 mg/kg fine particles of saline solution was used to establish modeling, every other day, three times. Thirty-two Wistar rats were randomly divided into normal group, model group and MQJWJD high-dose and low-dose groups, with eight rats in each group. Medication groups were given relevant medicine for gavage. The level of TNF-α in bronchoalveolar lavage fluid (BALF) was measured by ELISA. The expression of NF-κB protein in lung tissues was measured by immunohistochemistry. The histopathology of the lung injury was observed by light microscope. Results Compared with normal group, the level of TNF-α and the expression of NF-κB protein in the model group were higher than those in the normal group (P<0.01). Compared with model group, the level of TNF-α and the expression of NF-κB protein in MQJWJD low-dose and high-dose groups were lower than those in the model group (P<0.05, P<0.01). Pathological observation showed that, compared with normal group, model group showed intratracheal, alveolar and interstitial bacteria within a large number of fine particles calm, alveolar and pulmonary interstitial visible large amounts of phagocytic fine particles of macrophages and accompanied by more neutrophils and lymphocyte infiltration; Lung tissue pathological changes were significantly lighter in MQJWJD high-dose and low-dose groups than the model group. MQJWJD high-dose group showed mild inflammation, alveolar and pulmonary interstitial visible phagocytic fine particles of macrophages, a small amount of neutrophils and lymphocyte infiltration. Conclusion MQJWJD can reduce the pulmonary injury in rats induced by particulate matter and has protective effects on the rat model through decreasing the levels of TNF-α and the expressions of NF-κB protein in injured lung tissues.

OBJECTIVETo investigate the expression variation and significance of Skp2 and p27(kip1) during the proliferation of lymphoma cell line Jurkat cells.

METHODSThe binding of p27(kip1) and Skp2 in Jurkat cells were detected by immunoprecipitation. Jurkat cells were treated with serum starvation and release synchronization. The expression variation and subcellular localization of p27(kip1) and Skp2 were detected by subcellular fractionation, Western blot and double immunofluorescence labelling.

RESULTSThe results of immunoprecipitation suggested that p27(kip1) and Skp2 could bind each other in Jurkat cells. During the proliferation of Jurkat cells, the protein expression of p27(kip1) decreased and intranuclear p27(kip1) decreased significantly, while the Skp2 protein increased and cytoplasmic Skp2 increased significantly.

CONCLUSIONDuring the proliferation of Jurkat cells, the increased cytoplasmic synthesis of Skp2 may speed up p27(kip1) degradation via the ubiquitin-proteasome pathway, then intranuclear p27(kip1) decreases significantly, leading to an increased cell cycling activity.

Cell Nucleus ; metabolism ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cytoplasm ; metabolism ; Humans ; Jurkat Cells ; Lymphoma, B-Cell ; metabolism ; pathology ; Protein Binding ; S-Phase Kinase-Associated Proteins ; metabolism

OBJECTIVETo investigate the expression and relationship of p27(kip1) and its related molecules Jab1 and CRM1 during proliferation of lymphoma cells U937.

METHODSU937 cells were treated with serum starvation and release, and the effects of these treatments on the cell growth was tested with cell number counting. The expression and localization of p27(kip1), Jab1 and CRM1 in U937 cells were detected by Western blot, double immunolabelling and laser scanning confocal microscopy.

RESULTSThe growth of U937 cells was blocked by serum starvation. The total protein of p27(kip1) was increased while Ser10-phosphorylated p27(kip1) -related molecules Jab1 and CRM1 were decreased. Meanwhile, the location of p27(kip1) was changed from cytoplasm into nuclei. After serum release, the location of p27(kip1) expression reappeared in the cytoplasm again.

CONCLUSIONDuring the proliferation process of lymphoma U937 cells, Jab1 and CRM1 may influence the location and expression of p27kip1, and may participate in regulation of growth of NHL cells.

COP9 Signalosome Complex ; Cell Culture Techniques ; Cell Nucleus ; metabolism ; Cell Proliferation ; Culture Media, Serum-Free ; pharmacology ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cytoplasm ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Karyopherins ; metabolism ; Peptide Hydrolases ; metabolism ; Receptors, Cytoplasmic and Nuclear ; metabolism ; U937 Cells