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 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.

[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.

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 An epidemiological study was conducted to investigate the prevalence of female breast disease and risk factors in Tongzhou District of Beijing,providing objective data to prophylaxis of the disease in this area.Methods A questionnaire survey was done in 1 575 females aged 18-60 years in Tongzhou district of Beijing by a cluster random sampling method.Clinical breast examination,near- infrared scanning,colour ultrasound,X-ray and other adjunctive examinations were performed in these subjects for diagnosis of the disease.The subjects were divided into breast disease group and normal control group.And then a case-control study was conducted for risk factor analysis of female breast disease.Results Breast disease was found in 303 of 1 575 females,most in 30-50 years,the incidence being 19.2％.Breast hyperplasia was found in 270 females,breast fibroma in 5,other benign breast disease in 25,and breast cancer in 3,Breast disease was found related to education level,menopause time,history of breast hyperplasia and annoyance.Conclusions Female breast disease is related to age,education and menopause.History of breast hyperplasia and negative emotion are risk factora of the disease.Attention should be paid to breast health care and healthy life style,decreasing incidence of the disease.

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.

Objective To analyze and discuss the possible reasons of the bone resorption beneath the prostheses after chin augmentation.Methods Twelve patients were admitted to our department for further correction after chin augmentation with materials.The bone resorption was observed through the clinical research and X-ray examination.Results All the patients were underwent the removal of the materials,genioplasty was performed in 8 patients,and two patients were treated by chin augmentation with polyethylene.All the patients were satisfied with their facial contouring.Mild bone resorption was found in seven patients (depth of bone resorption ≤2 mm),in which five patients were used with silicone materials,two patients were performed with expanded polytetrafluoroethylene implants.Moderate bone resorption was seen in three cases.All of them were used with silicone implants (2 mm ＜ depth of bone resorption ≤4 mm).Severe bone resorption happened in two patients (depth of bone resorption ＞4 mm).One was used with silicone implant,and the other one was carried out with expanded polytetrafluoroethylene implant.Conclusions The imbalance among mentalis muscle,materials and underlying bone might be one of the key reasons.Thus for mild and moderate microgenia cases,chin augmentation with material is suitable,while long-term fellow-up study is necessary.But for the cases of severe mirogenia or microgenia and micrognathia with dentofacial deformity or mentalis muscle hyperactivity,genioplasty might be performed as well to correct their deformities.