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.

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.

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.

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.

The proliferation of MG63 cells under high glucose intervened with metformin was measured by CCK-8 assay.The activity of intracellular alkaline phosphatase (ALP) was detected by SFBC assay.The expression of related genes was detected by RT-PCR.Metformin promoted the proliferation of MG63 cells (P＜0.01),the activity of ALP was increased (P＜0.05).The expressions of collagen Ⅰ and osteocalcin were up-regulated while the expressions of matrix metalloproteinase (MMP)-1,MMP-2,MMP-3,and MMP-8 were down-regulated (P＜0.05).The results indicate that metformin may protect osteoblast,via promoting osteoblast proliferation,differentiation and mineralization,as well as by inhibiting degradation of osteoblast extracellular collagen matrix.

Objective To study the clinical value of mammography and ultrasound needle localization biopsy technique in clinical nonpalpable breast disease.Methods 148 patients with negative clinical palpation were operated only because of the mammography or ultrasound findings of lesions.They were undergone localized excision biopsy and derived definite pathological diagnosis.By histopathology diagnosis,benign and malignant lesions,lesion size,complete resection rate were statistically analyzed.Results 21 cases in 148 cases were malignant lesions(14.2％),which were all early breast cancer.The other 127 cases were benign breast disease(85.8％).147 cases were successfully resected at the first time and 1 case was successfully resected at the second time.Conclusion Mammography and Ultrasound fine needle localized biopsy in nonpalpable breast disease diagnosis and treatment has great significance,which can raise early diagnosis rate of breast cancer without mass,and this method is simple and easy for promotion.

Objective Primitive neuroectodermal tumor (PNET) is a rare malignant small round cell tumor .This paper aimed to study the clinical and pathological features of primary pulmonary primitive neuroectodermal tumor . Methods We collected 2 cases of primary pulmonary PNET to review the clinical and pathological features .Immunohistochemical staining was used to detect immune mark-ers, and fluorescence in situ hybridization (FISH) was applied to detect EWS translocation. Results 2 patients were aged 33 years and 17 years.Microscopically, the tumor cell was composed of single small round cells in diffusion or in distribution of sheets or beams , with scant cytoplasm , oval or spindle-shaped nucleus , high mitotic count .Irregular tumor necrosis scattered in the tumor along with visi-ble rosette structure.Immunohistochemical study showed that the tumor cells were positive for CD 99, FLI-1 and Syn, while CKpan, EMA, Desmin, CgA, TTF1, CD34 were negative.EWS/FLI1 translocations were detected positive in both the cases .2 patients died 7 months and 32 months after operation , respectively . Conclusion Primary pulmonary PNET is rare , so the selection of appropriate im-mune markers (CD99, FLI-1, Syn) and FISH for the detection of EWS translocation helps to improve the accuracy of diagnosis .

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.