Objective To determine the expression of aquaporin-1,3,8,9 mRNA (AQP-1,3,8, 9) in amniotic membranes in pregnant women with polyhydramnios. Methods Amniotic membranes were collected from women who presented with either polyhydramnios (n= 5)or normal amniotic fluid volume (control, n= 5) underwent elective cesarean sections at term. The AQP-1,3,8,9 mRNA expression were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Results The expression of AQP-9 mRNA on fetal membranes were significantly higher in polyhydramnios groups (1. 1403±0. 0831) than that of control (0. 5903±0. 1909) (P = 0. 002), although the expression of AQP-1, 3 and 8 showed no significant difference between the two groups (P= 0. 972, 0. 242,0. 608, respectively). Conclusions AQP-9 may play an important role in maintaining the volume and the balance of different components of amniotic fluid in polyhydramnios cases.

Objective To establish a method for the determination of alum residual amount in Rhizoma Pinelliae Preparata from different companies,and to provide a scientific evidence for quality evaluation of Rhizoma Pinelliae Preparata.Methods Na2EDTA titration method was applied for the quantitative analysis.Results There were obvious differences in residual quantity of alum in Rhizoma Pinelliae Preparata from different companies.Conclusion The method is simple,reproducible and accurate,it can be used for the determination of alum residual amount in Rhizoma Pinelliae Preparata.

To evaluate the effects of recombinant human stem cell factor on mobilization of peripheral blood stem cells, 30 monkeys were divided into control, rhG CSF 10?g?kg -1 ?d -1 (?KD), rhSCF 50?KD, rhG CSF 10?KD+rhSCF 20?KD, rhG CSF 10?KD+rhSCF 50?KD and rhG CSF 10?KD+rhSCF 125?KD groups. The monkeys were given subcutaneous injections of rhSCF for 8 days and/or rhG CSF for 5 days. The highest value of leukocyte in rhG CSF and rhSCF groups was 206% and 200% respectively, and that of rhG CSF+rhSCF groups was 280% to 310%. CFU GM of peripheral blood was up to 1 1, 1 3 and 5 8～7 9 times respectively after the mobilization. CD34 positive cell number of joint mobilization group was 20, 24 and 35 times respectively of that before mobilization. rhSCF can mobilize the effect of peripheral blood stem cells, and the effect of joint rhSCF+rhG CSF is better.

Objective To observe the curative effect of rhG-CSF or rhG-CSF+rhSCF mobilized peripheral blood stem cells (PBSC) on rhesus monkeys after irradiation of 7.0Gy ? ray.MethodsFifteen healthy adult rhesus monkeys were divided into placebo control, rhG-CSF, and rhG-CSF+rhSCF mobilized groups. rhSCF 20?g?kg -1 ?d -1 (?KD) was administered for 8 consecutive days. Then from the 4th day to the 8th day, animals of both rhG-CSF and rhG-CSF+rhSCF mobilized groups were also given rhG-CSF 10?KD. The placebo control animals were given 0.9% sodium chloride of the same volume per kilogram. On day 0 of the irradiation, PBSCs were collected from all subjects. 3～4 hrs after irradiation, all animals received autologous PBSC infusion. ResultsFrom day 21 to day 29 after TBI, rhG-CSF+rhSCF mobilized group showed higher WBC counts compared to placebo control and rhG-CSF mobilized group. Platelet counts of both mobilized groups recovered more quickly than those of placebo control. Bone marrow nucleated cells culture demonstrated that rhG-CSF and rhG-CSF+rhSCF mobilized PBSC had stimulated bone marrow nucleated cells to form more CFUs. Histopathological evaluation showed the number of hematopoietic cells in the bone marrow of rhG-CSF mobilized group was greater than those in placebo control but less than those in rhG-CSF+rhSCF mobilized group. ConclusionThe infusion of rhG-CSF or rhG-CSF+rhSCF mobilized PBSC can improve the hematopoietic recovery in rhesus monkeys after 7.0Gy ? ray irradiation. The combined use of two mobilizing factors gives better result than using single factor.

BACKGROUND: Placenta and fetal membrane play an important role In maternal-fetal homeostasis. However, the molecular and cellular mechanisms underlying water transfer across placenta and amniotic membrane remain unknown. It is hypothesized that maternal-fetal fluid exchanges via aquaporin (AQP) water channels in the placenta and fetal membrane.OBJECTIVE: To investigate AQP8 protein expression in normal human placenta and fetal membrane.DESIGN, TIME AND SETTING: A control observation was performed at the Central Laboratory of Guangzhou Medical College from July to December 2005.MATERIALS: Human placenta and fetal membrane tissues from 5 elective cesarean section deliveries of normal term pregnancies (range 37-42 weeks) were studied. Maternal age averaged (27?) years old. Experimental protocol was approved by the Hospital's Ethics Committee.METHODS: Thirty minutes after delivery, fetal membrane and placenta were dissected and washed with sterile physiological saline. Some were frozen at -80?, and the remaining tissues were fixed for 24-48 hours with 10% neutral formalin and paraffin embedded for immunohistochemical staining.MAIN OUTCOME MEASURES: AQP8 expression and distribution in human placenta and fetal membrane were detected by the reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blotting analysis.RESULTS: RT-PCR results showed that AQP8 mRNA was expressed in both placenta and fetal membrane tissues. Western blotting analysis also yielded positive results in placenta and fetal membrane with a specific band site at approximately 45 000.Immunohistochemistry results revealed that AQP8 protein was expressed in placental syncytiotrophoblasts, amniotic epithelial cells, and chorion cytotrophoblasts.CONCLUSION: At protein level, AQP8 is expressed in placental syncytiotrophoblasts, amniotic epithelial cells, and chorion cytotrophoblasts.

Objective To investigate the serum levels of CD147 and MMP-2 and the relationship between these proteins and colorectal tumor differentiation, Dukes stage. Methods Serum levels of CD147 and MMP-2 in 44 patients with colorectal carcinoma,28 polyp intestinal patients and 36 normal subjects were measured by ELISA. Results Serum levels of CD147(103.59±40.74pg/ml)and MMP-2(14.92±2.02ng/ml)in the cancer patients were significantly higher than those of the polyp intestinal(66.27±16.91;10:96±1.71)and normal control(63.84±18.71;10.49±1.61)(P＜0.05).The levels were associated with tumor differentiation and Dukes stage. Patients with poor differentiation had significantly higher levels of MMP-2 than those of patients with well differentiation. And Dukes C and D stage tumors had significantly higher levels of CD147 than Dukes A and B stage tumors(P＜0.05).The levels of CD147 and MMP-2 declined remarkably after one month of radical operation. Conversely, it decreased illegibly after palliative operation. The serum levels of CD147were positively correlated with MMP-2 in patients with colorectal cancer. Conclusion Elevated CD147 and MMP-2 serum levels are associated with tumor differentiation, invasion, metastasis. Dynamic alterations of serum CD147 and MMP-2 may be used as the indicators of diagnosis, the choice of operative method and the judgment of prognosis in patients with colorectal cancer.

Rapid detection of pathogenic microorganisms is important to the prevention and control of diseases.Com-pared with traditional approaches, electrochemical DNA biosensors present great advantages in promising rapid, portable, sensitive and cost-saving detection of pathogens.In this review, the working principle of electrochemical DNA biosensors and the progress in detection of pathogens is introduced, the latest developments of DNA tetrahedron structure and new nano materials in electrochemical DNA biosensors are reviewed, and the challenges to and prospects of development in this field are also discussed.

Objective To observe and identify the impact of a type of macro?pore bone block bioactive glass on osteo?blast in vitro. Methods Extract fluid of new bioactive glass was prepared withα?MEM culture medium as the bioactive medium group. And the concentrations of different ions were detected with Inductively Coupled Plasma?Atomic Emission Spectrometry in bioactive medium group andα?MEM medium group. MC3T3?E1 cells cultured in bioactive medium group were considered as ex?perimental group and cells cultured inα?MEM medium as control group. Giemsa and immunofluorescence staining was performed to detect the cell numbers, the karyoplasmic ratio and the average fluorescence intensity per cell. Cell proliferation and viability in different groups were detected by cell cycle analysis, MTT assay and BrdU assay, respectively. Total RNAs of cells in different groups were extracted and the expressions of ALP, OCN and collagenⅠwere measured by quantitative real time PCR. ALP stain?ing and alizarin red staining were performed to assess the differentiation and mineralization of MC3TC?E1 cells in different groups. Results The concentrations of Si and F were 40.02 ± 0.67 mg/L and 0.02 ± 0.001 mg/L in bioactive medium group, higher than 2.02±0.01 mg/L and 0.00 mg/L inα?MEM solution, and the concentration of Ca was lower than that inα?MEM solution. The con?centration of P and Na had no difference. In Giemsa staining, the cell number in 400 times field under a microscope was 106.0 ± 6.025 in bioactive medium group and 40.20 ± 3.639 inα?MEM medium group. In the immunofluorescence of vinculin, the karyo?plasmic ratio and the expression of vinculin were higher in bioactive medium group (40.85±5.720, 0.050 88±0.021 78) than inα? MEM medium group (21.93 ± 4.137, 0.023 60 ± 0.003 18). In cell cycle analysis, the proportion of cells retained in S and G2/M phase in the bioactive medium group was more than that in theα?MEM medium group after 72 hours of cell culture. In the BrdU and MTT assay, MC3T3?E1 cells in bioactive medium group both showed a higher proliferation rate with statistical significance. In MC3T3?E1 cells cultured with the bioactive medium, the expressions of osteogenesis?related genes were higher than those cultured with ordinaryα?MEM solution;in the ALP staining and alizarin red staining, the expression of ALP and the mineralization rate were higher in bioactive medium group (1.328%±0.015 36%, 2.953%±0.536 3%) than inα?MEM medium group (0.979%±0.030 59%, 1.000%±0.208 1%). Conclusion The bioactive medium promotes cell proliferation and osteoblastic differentiation of MC3T3?E1 cells, and has much more Si ions, which indicates that macro?pore bone block bioactive glass can promote cell proliferation and dif?ferentiation and has promising bioactivity.

Objective To detect the expression levels of the miR-205 in lung cancer tissue and A549 cells and its targeted gene YES1 using qRT-PCR and dual fluorescence protein repoter assay system,and to explore the possible mechanism of miR-205 to inhibit the proliferation of lung cancer A549 cells.Methods The expression levels of miR-205 in 10 cases of lung cancer tissue and adjacent normal lung tissue were detected with qRT-PCR.The cell growth curve and colony formation assay were used to determine the proliferation rate of A549 cells after transfected by miR-205 mimics and control mimics.The sequences of YES1 3′UTR (untranslated region)and mutation target sites of YES1 3′UTR were inserted into the plasmid which expressed green fluorescence protein (pcDNA3/EGFP) respectively to construct the green fluorescence protein plasmids of YES1-3′UTR and mut-YES1-3′UTR. There were six groups in the study:YES1-3′UTR, YES1-3′UTR and miR-205 mimics, YES1-3′UTR and control mimics,mut-YES1-3′UTR, mut-YES1-3′UTR and miR-205 mimics, mut-YES1-3′UTR and control mimics;after the plasmids expressed red fluorescent protein (pDsRed2-N1 )were cotransfected into A549 cells,the extracted protein was detected with fluorescence spectrophotometer.Results Compared with adjacent normal lung tissue,the expression levels of miR-205 in lung cancer tissue and A549 cells were decreased (P<0.05 );the proliferation rate of A549 cells in miR-205 mimics group was lower than that in control mimics group (P<0.05). The fluorescence protein expression level in YES1-3′UTR and miR-205 mimics co-transfected group was lower than that in YES1-3′UTR and control mimics co-transfected group, the difference was statistically significant (P<0.01).The number of cell colony formation of A549 cells in highly expressed YES1 group was higher than that in cell control group (P<0.05).Conclusion MiR-205 may inhibit the proliferation of A549 cells through regulating of the expression of YES1 directly.miR-205 and YES1 are potential therapeutic targets for the biological treatment of tumor.

Objective To explore the dynamic changes and clinical significance of relative pituitary hormones in children after craniocerebral injury.Methods The quantitative analysis and dynamic observation were performed in 125 children after craniocerebral injury and 20 voluntary healthy children of relative pituitary hormones including serum prolactin(PRL),cortisol(PTC),three free triiodothyronine (FT3),free thyroxine (FT4),thyroid stimulating hormone (TSH),growth hormone (GH) by applying electrochemical luminescence method.Tbe hormone variational characteristics were analyzed according to posttraumatic time,Glasgow Coma Scale(GCS) scores on admission and Glasgow Outcome Scale (GOS) scores on discharge,and the relationship between hormone variational characteristics of 58 cases was followed up over 2 years and the activities of daily living (ADL) were also investigated.Results The serum PRL was significantly increased on the first,third and fifth day compared with the healthy control group (P =0.000 0,0.000 0,0.006 7),respectively.There was significant difference between mild,moderate and severe groups within 30 days after suffering from craniocerebral injury (P ＜ 0.05).PTC was heavily increased within 3 days,and significant difference existed among mild and moderate groups mild and severe groups (all P ＜ 0.05) ; TSH,FT3,FT4 decreased slightly after injury and gradually rose in later;GH change wasn't significant;and the larger variation of relative pituitary hormones was responsible for lower GCS scores;FT3,FT4,TSH,and GH decreased in different degrees,which were found in parts of children with craniocerebral injury,and the significant difference of serum PRL existed between GOS scores 4-5 and GOS scores 1-3 groups (P =0.000 1).Conclusions The changes of relative pituitary hormones were associated with the posttraumatic time and the severity of craniocerebral injury.The PRL in serum can aid in prediction of outcome for the children with craniocerebral injury.