This study is to establish a simple and practical co-culture method of cortical neurons and astrocytes of rats. The cortex of the new-born SD rats was digested by 0.125% pancreatic enzyme, and the differential adherence was applied to obtain the mixed cell suspension of neurons and astrocytes. A low concentration of cytarabine was used to inhibit the astrocytes in a moderate way to get neuronal and astrocyte co-culture. The morphological characteristics of the cells in different times were observed under the inverted microscope. The cells began to adhere the wall 2 h after the inoculation. Neurons and astrocytes grew in a good condition under the inverted microscope 9 days after the inoculation. The results of the immunofluorescence staining and Rosenfeld's staining indicated that the co-culture of neurons and astrocytes was successful and the ratio of neurons and astrocytes was close to 1:1. A new neurons and astrocytes co-culture method, which is simple and convenient, was successfully established. It will be an efficient method for the related researches about neuronal and astrocyte co-culture in vitro.

OBJECTIVETo study the possible relationship between tumor necrosis factor (TNF)-alpha-238G/A gene polymorphism and the susceptibility to intrauterine HBV infection.

METHODSTwo hundred and fifty-six children, including 130 infants born to HBsAg positive mothers were divided into two groups: forty-five children with intrauterine HBV infection (group I) and 85 children without intrauterine HBV infection (group II), with a control group of 126. TNF-alpha-238G/A gene polymorphism was examined in all 256 children, by means of real-time quantitative fluorescent PCR.

RESULTSA significant difference of TNF-alpha-238A allele frequency was found between group I and group II (x2=6.797, P=0.009), and between group I and the controls group (x2=0.047, P=0.002), but there was no significant difference between group II and the control groups (x2=0.047, p=0.828).

CONCLUSIONThis study found that genetic polymorphism of tumor necrosis factor-a was associated with intrauterine HBV infection

Adult ; Child ; Disease Susceptibility ; Female ; Hepatitis B ; transmission ; Humans ; Infant ; Infectious Disease Transmission, Vertical ; Polymorphism, Genetic ; Tumor Necrosis Factor-alpha ; Tumor Necrosis Factors ; genetics

OBJECTIVETo detect the expression of CD133 in human tongue squamous cell carcinoma Tea8113 cell line and observe proliferation ability of CD133 groups in vitro.

METHODSLimiting dilution assay was employed to observe the proliferating character of Tca8113 single cell in vitro. The ability of growing as cancer spheroids was observed with ultra-low attachment plates. The flow cytometry was used to detect the expression of putative tumor-initiating cell marker CD133 in human tongue squamous cell carcinoma Tca8113 cell line. The selective technique of immunomagnetic beads was applied to purify CD133 tumor cells, CD133 tumor cells were cultured and their ability of proliferation were observed in vitro.

RESULTSAfter 12 days, the result of single cell culture in vitro revealed that about 5.23% of cultured Tca8113 cells possessed the capacity of continue proliferation. The cells line fromed floating clusters with one week of passaging cells into non-adherent plates. Approximately 0.95% of cells in Tca8113 cell line expressed CD133. Compared with CD133- cells and control Tca8113 cells, CD133+ cells demonstrated increased proliferation capacity. The proportion of CD133 cells decreased in culture as days passed. The percentage of CD133+ cells decreased from 92.45% to 1.62% in twelve days' culture.

CONCLUSIONTumor stem cells have the character of heterogenity and lower proportion of CD133 but higher ability of proliferation, and the diferentiation in human tongue squamous cell carcinoma Tca8113 cell line in vitro, CD133 may be one of makers for tumor-initiating cell of human tongue squamous cell carcinoma Tca8113 cell line.

Carcinoma, Squamous Cell ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Humans ; Tongue Neoplasms

OBJECTIVETo investigate the feasibility of using modified peroneal tendofascial compound flap with partial tendon of peroneal long muscle in lower leg refractory defect with Achilles tendon rupture.

METHODSBased on the application experience of peroneal tendofascial compound flap for lower leg refractory defects, partial tendon of peroneal long muscle was included in the flap to repair the Achilles tendon rupture.

RESULTS6 cases were treated. The maximum size of modified compound flap reached 5 cm x 5 cm. The maximal included tendon in the flap was 5 cm in length. All the flaps survived completely. Partial epidermis necrosis happened in one case between the skin graft at the donor site and the edge of the flap pedicle. The patients were followed up for 5 months to 2 years with satisfactory cosmetic and functional results.

CONCLUSIONSThe modified compound flap has reliable blood supply and can be easily transposed, leaving less morbidity at the donor site. It is very suitable for lower leg refractory defect with the Achilles tendon rupture.

Achilles Tendon ; injuries ; Adult ; Aged ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Surgical Flaps ; Tendon Injuries ; surgery ; Treatment Outcome

Phosphodiesterase 4 (PDE4) is an important member of the phosphodiesterase enzyme family that specifically catalyzes the hydrolysis of cyclic adenosine monophosphate (cAMP), activates the downstream phosphorylation cascade pathway by altering cAMP concentration, and is strongly associated with multiple diseases. Inhibition of PDE4 is clinically investigated as a therapeutic strategy in a broad range of disease areas, including respiratory system diseases, autoimmune disorders, central nervous system diseases, and dermatological conditions. However, the incidence of adverse reactions such as nausea and vomiting is relatively high in the marketed PDE4 inhibitors, which has stalled their clinical development. In this review, we provide an overview of the clinical progression and safety issues of the marketed PDE4 inhibitors. We also review the main causes underlying PDE4-mediated adverse effects by combining the structural analysis of the PDE4 protein, the mechanism of action of PDE4 inhibitors, and the related side effect mechanism research, aiming to provide a reference for the development of safe and effective PDE4 inhibitors.

OBJECTIVETo study the anatomy of peroneal tendofascial flap combined with adipofascial flap for the repair of heel tissue defects.

METHODSThe lower extremities of five cadavers (10 sides) were perfused with red latex, the blood supply of peroneal tendofascial flap and vicinity adipofascial flap were observed. The diameter, course, branches and location of the blood vessels were measured. Eight fresh cadavers (16 sides) were perfused with lead oxide-gelatine mixture. The covering fascia tissues of the lower extremities was obtained and photographed by X-ray. The vascular anastomosis and association of nutrient vessel of peroneal tendofascial flap and vicinity adipofascial flap were observed. Two adult lower extremities specimens (4 sides) were used to construct vessel diagrams for observation of the course, distribution and anastomosis of the vessels. Eight cases were treated successfully with theses flaps.

RESULTSThe blood supply of the combined fascial flap is multi-originated. For the area within 4 cm below and above the lateral malleolus cusp, the blood supply includes 2-5 branches from heel lateral artery with an average diameter of (0.5 +/- 0.2) mm, 1-2 branches from posterior lateral malleolus artery with an average diameter of (0.6 +/- 0.2) mm and 2-3 branches from the descending part of perforating branches of peroneal artery with an average diameter of (0.5 +/- 0.2) mm. The blood supply of area 4 cm above lateral malleolus cusp is 1-3 branches from intermuscular septum perforating branches of peroneal artery with an average diameter of (1.0 +/- 0.2) mm. These above branches are anastomosed each other and also send off many smaller branches to form vascular net around tendon. The fascial flaps and free skin grafts in eight patients were completely survived. All patients were followed up for 3-24 months, the donor and recipient sites were healed very well. The functional and cosmetic results were satisfactory.

CONCLUSIONSPeroneal tendofascial flap combined with adipofascial flap, with proximal pedicle or reverse distal pedicle, can be used to repair the defect at the lower leg and refractory small- and medium-sized defects at the heel.

Adipose Tissue ; anatomy & histology ; transplantation ; Adult ; Aged ; Fascia ; anatomy & histology ; transplantation ; Female ; Fibula ; Heel ; injuries ; surgery ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; anatomy & histology ; transplantation ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps

OBJECTIVETo explore the possible relationship between cytokines (TNF-alpha, IFN-gamma, IL-4 and IL-10), which were expressed abnormal quantity in the peripheral blood to intrauterine HBV infectious children, gene single nucleotide polymorphism (SNP) and susceptibility to HBV intrauterine infection.

METHODSA cross sectional study on molecular epidemiology was carried out. The subjects were selected from outpatients of the hepatitis B vaccine special clinics of our hospital. According to intrant criteria, children under high risk of HBV intrauterine infection were divided into immuno-failure group (group I) and immuno-effective group (group II) while children without high risk were included in the control group. Four gene SNP sites of TNF-alpha-238, IFN-gamma + 874, IL-4-590 and IL-10-1082 region were determined by real-time quantitative fluorescent PCR.

RESULTSSignificant differences of TNF-alpha-238 A allele frequency were found between group I and group II (chi(2) = 6.797, P < 0.05) as well as between group I and control group (chi(2) = 9.513, P < 0.05). No evident difference of TNF-alpha-238 A was found between group II and control group (chi(2) = 0.047, P > 0.05). Significant differences of IFN-gamma + 874 A allele frequency were found between group I and group II (chi(2) = 7.238, P < 0.05), and between group I and the controls (chi(2) = 5.199, P < 0.05) but no significant difference was found between group II and control group (chi(2) = 0.602, P > 0.05). Significant differences of IL-4-590 C/T allele frequency were not found between group I and group II (chi(2) = 0.632, P > 0.05), group I and control group (chi(2) = 0.584, P > 0.05), or between group II and control group (chi(2) = 0.004, P > 0.05) respectively. Significant differences of IL-10-1082 G allele frequency were found between group II and group I (chi(2) = 10.359, P < 0.001), and between group II and the controls (chi(2) = 35.418, P < 0.001), but not found between group I and control group (chi(2) = 1.759, P > 0.05).

CONCLUSIONThis study suggested the possibility that TNF-alpha-238 A allele and IFN-gamma + 874 A allele were associated with HBV intrauterine infection. There was no evident relationship between IL-4-590 C/T allele SNP and susceptibility to HBV intrauterine infection, but the IL-10-1082 G allele seemed to be associated with preventive efficacy to HBV intrauterine infection.

Case-Control Studies ; Child ; Child, Preschool ; Cytokines ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Hepatitis B ; transmission ; Humans ; Infant ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Interferon-gamma ; genetics ; Interleukin-4 ; genetics ; Polymorphism, Genetic ; Pregnancy ; Pregnancy Complications, Infectious ; Retrospective Studies ; Risk Factors ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo explore the relationship between the expression of Survivin and Ki67 with prognosis of pancreatic endocrine tumors (PETs).

METHODSImmunohistochemistry for Survivin and Ki67 was performed in 25 cases of normal pancreatic tissues and 81 cases of PETs by tissue microarrays and to observe the expression and evaluate the relationship with prognosis.

RESULTS(1)The expression of Survivin and Ki67 in PETs was significantly higher than that in normal pancreatic tissues (P <0.01); (2)The expression of Survivin and Ki67 in PETs was correlated with tissue grading and the TNM-staging (P < 0.05), but not related with tumor size, location and functional status. In addition, the expression of nuclear Survivin was association with lymph node metastasis (P < 0.05). (3)The high expression of Ki67 was related with the expression of nuclear Survivin, but not related with the expression of cytoplasmic Survivin.

CONCLUSIONSurvivin and Ki67 were both expressed in PETs, which were closely related to the clinical pathological characteristics. They could be used as new indicators in the evaluation of prognosis of PETs. The expression of Survivin in nucleus had more diagnostic significance than that in cytoplasm, and that could be highly correlated with lymph node metastasis, which would be used as a new marker of poor prognosis.

Biomarkers, Tumor ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; metabolism ; Ki-67 Antigen ; metabolism ; Lymphatic Metastasis ; Neoplasm Staging ; Pancreatic Neoplasms ; diagnosis ; Prognosis

OBJECTIVETo investigate the application and therapeutic effect of modified neurovascular flaps nourished by sural nerve and small saphenous venous, or saphenous nerve and great saphenous venous.

METHODSAccording to the anatomy and recipient area, we made some modifications for the two neurovascular flaps. 1) The small and great saphenous venous were ligated or anastomosed with the venous in recipient area; 2) The flap was delayed for 8 - 14 days if the donor area was above the middle and upper one third of leg; 3) The sural nerve and its branch were anastomosed to the nerve in recipient site to restore the sense of flap; 4) the lower turning point could be designed just 3 cm above the point of internal and external malleolus; 5) Partial gastrocnemius or soleus muscle could be included in the flap if needed; 6) Reversed cross leg flap was considered if the adjacent flap in the affected side could not be used. Sometimes, we adopted more than one modification in one patients.

RESULTSFrom Sep, 1994 to Apr, 2007, 63 cases were treated, including 51 cases of sural and 12 cases of saphenous neurovascular flaps. Partial necrosis (about 1 - 2 cm width) happened in the distal end of flaps in 2 cases which healed after dress changing. The patients were followed up for 3 months to 2 years with satisfied functional and cosmetic results.

CONCLUSIONSThese two modified flaps have reliable blood supply and survival rate. The operation is easily performed and very practical. The flaps, or combined with other methods can repair about 70% of the refractory defects in legs.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Lower Extremity ; innervation ; Male ; Middle Aged ; Skin Transplantation ; methods ; Surgical Flaps ; blood supply ; innervation ; Veins ; surgery ; Young Adult

BACKGROUNDThe influences of genomic background are confirmed in more diseases. Immunologic tolerance after intrauterine infection of hepatitis B virus is considered to occur in T cells. Cytokines work effectively in eliminating virus by immune system after hepatitis B virus infection. To explore the relationship between cytokines (tumor necrosis factor-alpha, interferon-gamma, interleukin-4 and interleukin-10), which expressed abnormal quantity in the peripheral blood to intrauterine hepatitis B virus infectious children, gene single nucleotide polymorphism (SNP) and susceptibility to hepatitis B virus intrauterine infection.

METHODSThis is a cross sectional study of molecular clinical epidemiology. The subjects in this study were selected from outpatients of hepatitis B vaccine follow-up special clinics of our hospital in the period. According to intrant criteria, the high risk children of hepatitis B virus (HBV) intrauterine infection were divided into immune failure group (group I); and immune effective group (group II) and non high risk children belonged to the control group. Four gene SNP sites of TNF-alpha -238, IFN-gamma +874, IL-4 -590 and IL-10 -1082 were determined by real-time quantitative fluorescent polymerase chain reaction (PCR).

RESULTSThe significant differences of TNF-alpha -238 A allele frequency were found between group I and group II (chi(2) = 6.797, P < 0.05) and between group I and the control group (chi(2) = 9.513, P < 0.05). No evident differences of TNF-alpha -238 A were found between group II and control group (chi(2) = 0.047, P > 0.05); the significant differences of IFN-gamma +874 A allele frequency were found between group I and group II (chi(2) = 7.238, P < 0.05), and between group I and the control group (chi(2) = 5.199, P < 0.05). No evident differences were found between group II and the control group (chi(2) = 0.602, P > 0.05); the significant differences of IL-4 -590 C/T allele frequency were not found between group I and group II (chi(2) = 0.632, P > 0.05), also group I and the control group (chi(2) = 0.584, P > 0.05), and the group II and the control group (chi(2) = 0.004, P > 0.05) respectively; The significant differences of IL-10 -1082 G allele frequency were found between group II and group I (chi(2) = 10.359, P < 0.001), and between group II and the controls (chi(2) = 35.418, P < 0.001), but the significant differences were not found between group I and the control group (chi(2) = 1.759, P > 0.05).

CONCLUSIONSThis study suggested the possibility that the TNF-alpha -238 A allele and IFN-gamma +874 A allele were associated with HBV intrauterine infection. There was no evident relationship between IL-4 -590 C/T allele SNP and susceptibility to HBV intrauterine infection, but the IL-10 -1082 G allele was associated with preventive efficacy to HBV intrauterine infection.

Cross-Sectional Studies ; Cytokines ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Hepatitis B ; genetics ; transmission ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Interferon-gamma ; genetics ; Interleukin-10 ; genetics ; Interleukin-4 ; genetics ; Male ; Pregnancy ; Tumor Necrosis Factor-alpha ; genetics