A novel PCR-based mutagenesis method was reported, in which there is no need to purify megaprimers or design a special flanking primer. This method used one mutagenic primer and two sequencing primers (T_m≤58℃) as flanking primers. After first round PCR, 12.5 μl first PCR production was directly added into 50 μl second PCR system as template and megaprimer, and 10 rounds of asymmetrical PCR at high temperature of annealing (68 ℃ ) was to add in initiation of second PCR. This additional step greatly has increased the efficiency of mutagenesis via 600 bp or 800 bp long megaprimer. The results demonstrated that this method can achieve high fidelity, 97%～ 98% efficiency, high yield.

Objective To investigate the effect of recombinant human growth hormone on serum levels of insulin like growth factor 1(IGF-1) and insulin-like growth factor binding protein 3(IGFBP-3)in children with short stature.Methods 48 children were diagnosed with idiopathic short stature were collected.All children were randomly divided into control group and experimental group ,24 cases in each group.Children in control group received nutritional therapy, children in experimental group were given recombinant human growth hormone on the basis of control group treatment, both group were treated for 12 month.After the treatment, the serum levels of IGF-1,IGFBP-3 and the height were detected in all children.Results Compared with control group post-treatment,,the serum level of IGF-1 was higher in experimental group ( P<0.05 );the serum level of IGFBP-3 was higher in experimental group ( P<0.05 );the height was higher in experimental group ( P<0.05 ) .Conclusion Recombinant human growth hormone can significantly increase the serum levels of IGF-1 and IGFBP-3 in children with idiopathic short stature, promote children growth, and have a guiding significance for clinical.

[Objective] To study the cure effect of Ganle Infusion on hepatitis-B of depressed liver and deficient spleen type.[Method] Randomly divide 60 cases into treatment group(Ganle Infusion) and control group(Diammonium Glycyrrhizinate capsule),observe concerned indexes change and clinical effects.[Result] For treatment group,the cure effect index was 93.3%;66.7% for control one;on liver improvement and serum hepatitis-B virus index negative-transfer rates,the treatment group was better than control one,with marked difference.[Conclusion] Ganle Infusion has marked cure effect on chronic hepatitis-B of the said type.

Juvenile granulose cell tumor is one of the rare sex cord stromal tumors of the ovary.This review summarizes the new progress in the genetic studies of the disease.

Objective To study the effect of 3,4-oxo-isopropylidene-shikimic acid (ISA) on transcription factor STAT1, STAT3 and NF-?B in human cervix cancer Hela cell. Methods The cell proliferation was assessed by MTT assay. The luciferase activity of transcription factor STAT1, STAT3 and NF-?B was determined by the Dual-Luciferase Reporter Assay System. Results ISA can not inhibited the growth of Hela cell. The luciferase activity of transcription factor NF-?B in Hela cells treated with ISA was inhibited, while the luciferase activities of transcription factor STAT1 and STAT3 were not inhibited. Conclusion ISA can inhibit inflammation, which may be related with suppression of NF-?B transcriptional activity.

In this paper an expert system for remote fault diagnosis in the ship lift was developed by analysis of the fault tree and combination with VPN. The fault tree was constructed based on the operation condition of the ship lift. The diagnosis model was constructed by hierarchical classification of the fault tree structure, and the inference mechanism was given. Logical structure of the fault diagnosis in the ship lift was proposed. The implementation of the expert system for remote fault diagnosis in the ship lift was discussed, and the expert system developed was realized on the VPN virtual network. The system was applied to the Gaobaozhou ship lift project, and it ran successfully.

Objective To assess the dose reduction potential of adaptive statistical iterative reconstruction(ASiR)and model-based iterative reconstruction(MBIR)in pelvic CT with a standard male phantom.Methods A Fluke Biomedical RANDO standard male phantom was scanned with discovery CT750 HD using different tube currents.CT images were reconstructed with FBP,50％ASiR and MBIR.The CT value,the image noise and the contrast-to-noise ratio(CNR)for the sacral vertebra relative to muscle were measured.The volume CT dose indexes(CTDIvo1)and dose-length product(DLP)were recorded.Results Compared with FBP,using 50％ASiR and MBIR had significant reduced image noise and greater CNR.The effective minimal tube currents for displaying sacral vertebra were 250 mA(FBP),180 mA(50％ASiR),and 100 mA(MBIR).With the similar image quality using FBP,the dose was reduced by 28.0％ and 59.9％ using 50％ASiR and MBIR,respectively.Conclusions Using advanced iterative algorithms can reduce image noise,improve CNR,and reduce the radiation dose in pelvic CT examination.

Purpose To investigate the clinicopathologic changes of the lung epithelial tumor with epithelioid granuloma. Methods E-leven cases of lung malignant tumor with epithelial epithelioid granuloma reaction were studied according to the histological changes of lung epithelial tumor, immunophenotype, acid-fast staining, and tuberculosis PCR test results combined with follow-up data. Results The mean age was 62. 3 years, and male to female ratio was 10∶ 1. The main clinical manifestations displayed a cough, chest pain, fever and hemoptysis or bloody sputum. Nine cases were malignant tumors, tuberculosis or inflammatory lesions was 1 case each in pre-vious surgery. The tumor with epithelioid granuloma formation of the 11 cases displayed in the same site ( mixed) . The histopathologic changes of the lesions showed a epithelioid or typical tuberculous granulomas formation, including 4 cases of adenocarcinoma, 5 cases of squamous cell carcinoma and 2 cases of neuroendocrine tumors. The acid-fast bacilli staining was positive in 3 cases;6 cases were positive for PCR detection of mycobacterium tuberculosis. 5 patients died within 2 years due to various causes. Conclusions The lung epithelial tumors with epithelioid granuloma is more commonly found in the middle-aged, The signs and symptoms of the patients are similar to primary epithelial malignant tumors of the lung . The biological behavior was progressive with poor prognosis. The diagnosis of the lesions depends on pathological examination. It is very important to pay more attention to the pathological examination, especially in lung puncture samples, and the differential diagnosis for a definate diagnosis and treatment.

Background and purpose: Reports showed that berbefine not only had an effect ofdetoxification and inflammation but also could induce the cell to apoptosis. Berberine may also inhibit the migration and metastasis of tumor. To investigate the effects and mechanism of berberine on migration and metastasis of A549 cells, the proliferation, mobility and adhesion of A549 cells were observed after incubation with berberine. Methods: The proliferation was determined by MTT. Wound healing assay and adhesion test were used to observe the mobility and adhesion rate. The mRNA and protein expressions of MMP2 and MMP9 were assessed by semi-quantitive RT-PCR and gelatin zymography, respectively. Results: Berberine could significantly inhibit the proliferation ofA549 within a certain range of treating time and dose (P<0.05). The mRNA and protein expression of MMP2 and MMP9 gene were decreased slightly by the treatment ofberberine (P<0.05), especially at dose of 100 μg/ml. Conclusion: Berberine can inhibit the migration and adhesion abilities of A549 cells, probably by down-regulating the expression and activation of MMP2 and MMP9.

Objective To explore the morphological, immunohistochemical characters and prognosis in one case of patients with cutaneous anaplastic large cell lymphoma complicated with acute myeloid leukemia (C-ALCL-AML). Methods The histopathology, immunohistochemical markers and follow-up information of one case of ALCL-AML was analyzed and the correlated literature was reviewed. Results The patient, 69 year-old, female, was initially present with shin lesion on one finger and abnormal myelogram. The histopathology of shin lesion showed that tumor cells were composed of large cells with abundant cytoplasm,the nuclei were large and irregular, and were infiltrated by Neutrophil and eosinophil. The CD30,CD3 and CD43 of tumor cells were positive, but ALK negative by immunohistochemical method. The number of WBC in peripheral blood was 15.5×109/L and 51 archaeocytes were in every 100 karyotes. Bone marrow aspiration detection showed that bone marrow was hyperplasia and the ratio of myeloblast was 78 %. This patient was diagnosed as C-ALCL-AML, partly differentiation type(AML-M2a). Conclusion C-ALCL-AML is very rare. Its diagnosis is dependent on clinical data, histopathology and immunohistochemical markers. The first choice of treatment is chemotherapy, but its prognosis is poor.