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.

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 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.

Objective To compare the impact of propofol and sevoflurane on thermoregulation in children undergoing orheopaedic surgery.Methods Sixty-eight children scheduled to undergo ortho-paedic surgery were randomly allocated to receive propofol (group P)and sevoflurane(group S)anes-thesia,34 cases in each group.Tympanic temperature was recorded 5 minutes before (T0 )and 5 min (T1 ),1 5 min (T2 ),30 min (T3 ),45 min (T4 ),60 min (T5 ),75 min (T6 ),90 min (T7 ),105 min (T8 )and 120 min (T9 )after anesthesia.Total fluid intake,duration of surgery,duration of anesthe-sia,the incidence of hypothermia,and the incidence of shivering were also recorded.Results Com-pared with T0 ,in both groups body temperature declined at T1-T8 .There was no difference between the two groups in total fluid intake,duration of surgery,duration of anesthesia and the incidence of shivering.Compared with group P,children in group S had a higher incidence of hypothermia(8 vs 1). Children in group S had lower temperature,which had statistical significance at T7 and T8 (P <0.05). Conclusion The core temperature of children undergoing orthopaedic surgery showed a trend of in-crease after the first fall in the surgery.Compared with propofol,sevoflurane anesthesia is more likely to lead to the incidence of hypothermia in children undergoing orthopaedic surgery in 90 min after in-duction of anesthesia.