Cholangiocarcinoma is a malignancy of biliary tract and its incidence has been increasing in recent years all over the world.Meanwhile,there have been advances in techniques for its diagnosis and treatment.Some new methods such as PET,endoscopic ultrasonography and scopic CT can promote the diagnostic rate in the early stage and they have been used for diagnosis and staging of tumors.For those patients with large cholangiocarcinoma in the liver,photodynamic therapy is an effective neoadjuvant treatment.For those patients with unresectable cholangiocarcinoma,organ transplantation might be employed. Classification of cholangiocarcinoma and study on its diagnosis are important for disease evaluation in patients with the disease.

[Objective]To explore the effect on spinal growth and devetopment at different month-age of immature rabbits with spinal fusion and fixation;and to explain the mechanism of crankshaft phenomenon after child scoliosis treatment.[Method]Twenty-four 6-weeks-old rabbits and 24 15-weeks-old rabbits were randomly divided into 2 groups.Various lengths of lumbar vertebral region were fused posteriorly and fixed with wires,and the Cobbs angle in the frontal and saglttal planes were measured after the surgery.[Result]The lumbar spine of each group became lordosis with different degree.It was most obvious in the group which were fused in 7 lumbar vertebraes at 6-month-week age.The multiple linear regression outcomes show significance among different month-age,different lengths of fusion region and the Cobbs angle.[Conclusion]In the imature rabbits,if their lumbar vertebrae were posteriorly fixed only may be appeared and getting serious gradually due to the potential growth of anterior spinal column.There has significant correlation among the degree of skelecton immature,fused segment length and spine lordosis.

[Objective]A new type of external frame for the treatment of club foot in three dimensions was reported.The characteristics of this frame and Illizarov frame were compared.[Method]The new frame was consisted of 3 couples with 6 metal rods.The rods have lengthening or compressing force on foot in three dimensions respectively through 4 Kire wires in tibia,calcaneus and tarsals.Sixty-two cases with 85 club feet followed more than 12 months were treated by the frame since Fabruary 2004.[Result]According to Gaeceau standerd and clinical appearance,function of the feet after the treatment,40 feet were excellent,40 feet were good,5 feet were improved,no poor case.[Conclusion]The new type of external frame is safe,effective and reliable for the treatment of club foot in three dimensions.Compared to Illizarov frame,it is more simple,comfortable,cheep and easy to be managed.

ObjectiveTo study the effect of anti-oncogene p15 and p16 on the proliferation of human cholangiocarcinoma cell line.MethodsThe cDNA of anti-oncogene p15 and p16 was constructed into pcDNA3-neo plasmid carrier. The human cholangiocarcinoma cell line QBC939 were transfected with the recombinants pcDNA3p15 and pcDNA3p16 using lipofectin, respectively. The expression products were analyzed by Western blot. Cell viability and death were measured with MTT assay. Cell cycle was determined by flow cytophotometry and the formation of cell clone was detected. Results The growth of QBC939 cells was inhibited. The flow cytophotometry verified p15 and p16 induced QBC939 cell G1 blockade. Conclusion Anti-oncogene p15 and p16 together lead to the inhibition of cell cycle.

Objective To explore the clinical significance of combined detection of C‐reactive protein (CRP)and white blood cells (WBC)in children with anemopyretic tonsillitis .Methods A total of 284 patients with anemopyretic tonsillitis were divided into ex‐cessive pattern group (131 cases) ,wind‐heat invasion group (153 cases) ,other 120 healthy children were recruited into control group .The WBC ,percentage of neutrophils (NEU % ) ,percentage of lymphocytes (LYM % ) and CRP were analyzed among the three groups .Results WBC and NEU% in excessive pattern group and wind‐heat invasion groups were higher than those in the control group (P< 0 .05) ,CRP in wind‐heat invasion group was higher than that in excessive pattern group and the control group (P< 0 .05) .CRP in the excessive pattern group were higher than that in the control group (P< 0 .05) .When CRP cut‐off value in wind‐heat invasion group was 12 .65 mg/L ,the sensitivity was 90 .6% ,specificity was 86 .3% .Conclusion The combined detection of WBC and CRP have clinical value in differential diagnosis of children with anemopyretic tonsillitis .

[Objective ] To investigate the improvement of the surgical treatment effect on cerebral palsy with spastic diplegia by performing modified surgeries since 2004. [Method ] Twenty-three cases of severe cerebral palsy with spastic diplegia were operated, 14 male, 9 female; age 3 ～ 12 years; the average age of 5.7 years; their main symptoms were unable to stand and walk independently, with severe hypermyotonia of both lower extremities, and a scissoring type of gait when standing, hip and knee flexing contracture, ankle and foot equinovarus. Physical examination: there was hypermyotonia of the iliopsoas, adductor musculus, hamstrings, triceps surae, anterior and posterior tibial muscle, flexor pollicis longus and flexor digitorum longus. The muscle tone was 3 ～4 grade according to Ashworth criterion. Treatment: The intramuscular tenotomy, fascial division or sliding elongation were performed, to lengthen the tendon of the joint flexion contracture, the lateral half of anterior tibial muscle tendon was transferred laterally to balance the muscle force at the foot. Finally, both lower extremities were fixed by a new designed external frame, to straigthen the articular genu, keep both ankles and feet in neutral position, both lower extremities abducting 30°, and to correct all joint deformities. Family rehabilitation programs started after the external frame was removed 6 weeks later. [Result] All cases were followed up in 1 ～3 years with the average 2. 2 years. The results were evaluated as excellent, good and ineffective, according to correction of the joint deformities, the ability to stand and walk, and parent's satisfaction to the treatment. Of them 21 cases were excellent, 2 cases good, and no ineffective case. [ Conclusion ] For the severe cerebral palsy with spastic diplegia in children, reasonable release of the tight tendons of the multi-joint flexion contracture, balance of muscle force at the foot, correction of all deformities and fixation of the lower extremities with the new external frame, and long term of family rehabilitation after the surgery, are more effective treatment.

Objective To investigate the changes in histological characteristics and collagen content in uterosacral and cardinal ligaments of pefimenopausal women in relation to relaxation of pelvic supports.Methods Twenty-eight subjects undergoing hysterectomies were selected,in which 14 cases were perimenopausal women with relaxation of pelvic support as the relaxation group and 14 women at perimenopansal age with leomyoma,cervical cancer,adenomyosis as the control group.Samples of cardinal ligaments and uterosacral ligaments were obtained at hysterectomies,and the tissues were shced and stained bv Masson's trichrome technique.Histological characteristics of the samples were studied and immunohistochemistry assay was applied to demonstrate the contents of collagen types I andⅢ.Results (1)The collagen in uterosacral ligaments and cardinal ligaments were stained blue by tlle MaSson's trichome technique.In comparison to the control group,the relaxation group had milder positive stains of the collagen and the stains were distributed in unequal intensities.Collagen content was arranged in loose pattern.Focal arrangement of the collagen was dense but fragmented.Collagen fibers were atrophic. (2)In immunohistochemistry assay and image analysis,collagen was positive in light to deep brown areas.In the relaxation group,positive units of collagen types I and III in cardinal ligaments were 13.8±2.1 and 9.6±2.4 respectively.Positive units of coHagen types I and Ⅲ of cardinal ligaments in the control group were 27.4 ±3.5 and 17.7 ±4.0 respectively.Difierences between these two groups were statistically significant (P

Objective To treat femur fracture in neonates using a new homemade harness designed and observe the effec of this harness. Methods Designed a new harness used in femur fracture in neonates,and used this harness in 7(8 femur fracture)csses,including 6 boys(7 femur fracture)and 1 girl,the age from 1 day to 12 days(average 4.7 days).Those cases included proximal thirds(2 cases)and middle thirds(6 cases)of femur fracture.The angle of fracture was from 44°to 83°(average 62.4°)before treatment. Results The angle of fracture was from 0°to 22°(average 14.0°)after treatment using the harness.Hespitalization was from 2 to 3 days.There were no skin sloughing or harness breaking off.All cases were followed up 6 to 36 months(average 21.3 months).All femur fracture healed in good alignment with leg-length discrepancy＜1 cm.Movement of hip and knee was normal. Conclusions The harness is a better method to treat femur fracture of the proximal and middle thirds in neonates.Advantages of it include simply operation,minimal hospitalization,minimal cost,and easy nursing.

Objective To explore the mechanism of 125I seed interstitial implantation-induced apoptosis of liver VX2 tumor cells in experimental rabbits, and to compare the effects of different radioactivity 125I seeds on the apoptosis and on the proliferation of tumor cells. Methods A total of 24 rabbit models with VX2 liver cancer were randomly and equally divided into 3 groups, and 125I seeds with different initial radioactivity were separately implanted into the rabbits of the three groups. 125I seeds of 0 mCi radioactivity were used in the control group (n=8), 125I seeds of 0.7 mCi radioactivity were used in the 0.7 mCi group (n=8) and 125I seeds of 1.0 mCi radioactivity were used in the 1.0 mCi group (n=8). The experimental rabbits were sacrificed at 5 weeks after the implantation; the tumor lesions were removed, and the effects of 125I seeds on the apoptosis and proliferation of tumor cells were determined. The tumor cell apoptosis rate, tumor growth-related factors, tumor growth factor expression protein and the influence of caspase-3 activity were evaluated. Results Regardless of their initial radioactivity, all the 125I seeds could make the tumor cell apoptosis rate increased, make Bcl-2 and VEGF expression level decreased, and make Bax expression increased, which were more obvious in 1.0 mCi group (P<0.05). The 125I seeds could increase the activity of caspase-3 within tumor tissue, but the difference between the 0.7 mCi group and the 1.0 mCi group was not significant (P>0.05). Conclusion The implanted 125I seeds can not only inhibit tumor’s growth through inducing apoptosis of tumor cells, but also inhibit tumor’s angiogenesis through influencing the expression of apoptosis-related gene and coding protein.

Aim To explore the effect of connexin 43 ( Cx 4 3 ) on acquired gefitinib-resistance in human non small cell lung cancer ( NSCLC ) . Methods HCC827 GR, a gefitinib-resistant ( GR) NSCLC cell lines from their parental cells was established by gradually in-creasing the concentration of gefitinib. Gefitinib effica-cy in HCC827 and HCC827 GR cells was detected by MTT assay. Expression of Cx43 mRNA in HCC827 and HCC827 GR cells was determined by RT-PCR. The protein expressions of Cx43 and phospho-Akt ( p-Akt) in these cells were detected by Western blot. The func-tional gap junction intercellular communication ( GJIC ) was measured by parachute assay. The cellular locali-zation of Cx43 protein was evaluated by immunofluores-cence staining. Results MTT assay showed less ge-fitinib cytotoxicity in HCC827 GR cells than that in their parental cells with IC50 of (10. 84 ± 0. 021) μmol ·L-1 versus (0. 07 ± 0. 019) μmol·L-1 , respective-ly. Moreover, both mRNA and protein expressions of Cx43 in HCC827 GR cells were significantly lower than those in HCC827 cells ( P<0. 05 ) . However, the p-Akt protein in HCC827 GR cells was obviously higher than that in HCC827 cells ( P<0. 05 ) . Furthermore, treatment with LY294002 caused a significant reduced p-Akt expression, but a significant increased Cx43 ex-pression in HCC827 GR cells. Moreover, no detecta-ble GJIC was found in HCC827 and their GR cells with or without RA ( a well-defined GJIC enhancer ) treat-ment. Immunofluorescence staining clearly showed that Cx43 protein accumulated in the cytoplasm of HCC827 and their GR cells. Conclusion The down-regulation of Cx43 expression in cytoplasm of HCC827 GR cells may contribute to the acquired gefitinib resistance in NSCLC cells by GJIC-independent activation of PI3 K/Akt signaling pathway.