Objective To observe the location of pelvic organs, the morphology and function of levator ani muscle (LAM) in primiparous women post vaginal delivery at 6 months postpartum using static and dynamic MRI, and investigate the correlation between LAM injury and pelvic organ prolapse (POP). Methods A perspective analysis of static and dynamic MRI was performed in fifty-one primiparous women post vaginal delivery at 6 months postpartum and thirty-five nulliparous women without experience of pregnancy and delivery as control group from June 2014 to January 2015. Previous pregnancy and abortion history, previous pelvic surgery and pelvic mass diseases were excluded. Cases with pelvic floor dysfunction symptoms were excluded from the control group. All of the women underwent static and dynamic MRI. The primiparous group was divided into two groups on presence or absence of POP on MRI findings:primiparous POP group and primiparous control group. The levatorani scoring system based on static MRI was used to characterize morphological changes of LAM into none, minor and major injury by the total score of bilateral LAM. A series of parameters including H line (the distance between the inferior margin of pubic symphysis to anorectal junction), M line (the perpendicular distance between the distal end of H line to pubococcygeal line), levator plate angle (LPA), iliococcygeal angle (ICA), and levator hiatus length and area were measured on static and dynamic MR images. Fisher exact test was performed to compare difference in distribution of the LAM injury between the primiparous group and control group, as well as the primiparous POP group and primiparous control group. Independent sample t test or Mann-Whitney test was used to compare difference in LAM parameters between the primiparous POP group and primiparous control group. Results In the 51 cases primiparous group, 44 cases showed none injury, whilst 5 cases with minor and 2 cases with major injury in the puborectal muscle. Thirty two cases showed none injury, whilst 10 cases with minor and 9 cases with major injury in the iliococcygeal muscle. In the 35 cases control group, none injury was shown in puborectal muscle, whilst 32 cases with none, 2 cases with minor and 1 case with major injury in the iliococcygeal muscle. There was no significant difference in the puborectal muscle injury between the two groups (P=0.203), and there was significant difference in the iliococcygeal muscle injury between the two groups (P<0.05). In the 24 cases primiparous POP group, 20 cases showed none injury, whilst 2 cases with minor and 2 cases with major injury in the puborectal muscle. Fourteen cases showed none injury, whilst 6 cases with minor and 4 cases with major injury in the iliococcygeal muscle. In the 27 cases primiparous control group, 24 cases showed none and 3 cases with minor injury in the puborectal muscle, whilst 18 cases with none, 4 cases with minor and 5 cases with major injury in the iliococcygeal muscle. There was no significant difference in the puborectal muscle injury and iliococcygeal muscle injury between the two groups (P=0.588 and 0.559, respectively). The LH during Valsalva status in primiparous POP group and primiparous control group were (6.7 ± 1.1) and (5.0 ± 0.6) cm, respectively, whilst the LHA was (41.6 ± 12.6) and (24.2 ± 5.5) cm2. There were significant difference between the corresponding groups (P=0.042 and 0.004, respectively). There was no significant difference between the corresponding groups of the other LAM parameters on static and dynamic MRI (all P>0.05). Conclusion Vaginal delivery may cause various degrees of LAM injury, the LAM functional deficiency were observed in primiparous women combined with POP.

OBJECTIVETo compare the therapeutic effect of Astragalus and Angelica Mixture (AAM) on treating CKD patients according to different CKD primary diseases, staging and TCM syndromes.

METHODSA multicentre, open-label, and self control clinical design was used, and thirty-two patients in line with inclusive criteria were recruited. Based on maintaining their previous basic CKD treatment, patients additionally took AAM (Astragalus and Angelica each 30 g), once a day, three months consisted of one therapeutic course. Serum creatinine (SCr), estimated glomerular filtration rate (eG- FR), 24 h urinary total protein (UTP), plasma albumin (ALB), hemoglobin (Hb), and changes of TCM syndrome factor integrals were compared before treatment, at the end of month 1, 2, and 3. The differences in the aforesaid indices were compared between CKD patients with different CKD primary diseases (chronic glomerulonephritis, chronic renal tubulointerstitial disease, hypertensive renal damage), different CKD stages (CKD 3 and CKD 4), and patients of qi-blood deficiency syndrome (QBDS) and non-QBDS.

RESULTSAAM could improve 78.12% (25/32) patients' renal function. Compared with before treatment, SCr decreased (12.08% +/- 10.11%), eGFR increased (21.14% +/- 18.55%), and ALB increased (2.76% +/- 1.97%) at the end of 3-month treatment (all P < 0.05). As for TCM syndrome factor integrals, compared with before treatment, the integrals for qi deficiency syndrome, blood deficiency syndrome, and yin deficiency syndrome decreased, while the integrals for dampness heat syndrome and turbid-toxin syndrome increased (all P < 0.05). There was no obvious difference in all indices except the integral for hypertensive renal damage patients of yin deficiency syndrome (P > 0.05). The SCr decreasing percent was 19.82% +/- 8.30% for patients of non-QBDS and 5.24% +/- 10.75% for patients of QBDS. The latter was higher with statistical difference (P < 0.05). As for TCM syndrome factor integrals, the integral differences of qi deficiency and blood deficiency were obviously higher in patients of QBDS, when compared with patients of non-QBDS (P < 0.05).

CONCLUSIONAAM could improve the renal function of CKD patients, elevate their ALB levels, and ameliorate associated qi deficiency syndrome, blood deficiency syndrome, and yin deficiency syndrome, especially for CKD patients of QBDS.

Adolescent ; Adult ; Aged ; Angelica ; Astragalus Plant ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Phytotherapy ; Renal Insufficiency, Chronic ; drug therapy ; Treatment Outcome ; Yin Deficiency ; drug therapy ; Young Adult

Objective To construct a nucleic acid vaccine expressing H.pylori HpaA and inter- leukin-2 gene and to identify the immunogenicity of the vaccine proteins in vitro and protection in vivo. Methods The H paA gene fragment was amplified by polymerase chain reaction(PCR) from the genomic DNA of the standard H.pylori strain 17874.Mouse interlukin(IL)-2 gene was amplified from pClneo- IL-2.The HpaA and IL-2 were cloned into pUCmT vector.After DNA sequences of the amplified HpaA gene and IL-2 were confirmed,both were cloned into the eukaryotic expression vector pIRES through a serial of enzyme digestion and ligation reactions.The recombinant plasmids were screened by PCR and restriction enzyme digestion.Then,recombinant pIRES-HpsA-IL-2 was transfected to COS-7 cells using Lipofectamine~(TM)2000.The immunogenicity of HpaA and IL-2 protein was detected by SDS- PAGE and Western blot.The recombinant plasmids were transformed to LB5000 and then to final host SL7207.The recombinant strains were passaged repeatedly.The mice were challenged with H.pylori after 4 weeks of inoculation of nucleic acid vaccine.H.pylori infection was detected by rapid urease test.Results The amplified HpaA gene fragment and IL-2 were confirmed by sequence analysis.The eukaryotic expression vector plRES and the pIRES-HpaA-IL-2 construction were confirmed by PCR and restriction digestion.The expressions of HpaA(30 000) and IL-2(14 000)protein by pIRES-HpaA-IL- 2 were detected by Western blot.The in vivo study showed that 75.0% and 58.4% of mice vaccinated by HpaA-IL-2 and HpaA,respectively,were protected anaigst H.pylory infection,which was signifi- cant different in comparison with PBS control (P

Objective To evaluate the value of 18F-FDG PET/CT in preoperative diagnosis and staging of suspected extrahepatic cholangiocarcinoma (EHCC).Methods The clinical data of 116 patients (72 males,44 females;age range 26-89 years) with suspected EHCC from January 2013 to October 2014 were retrospectively analyzed.All patients received preoperative whole body 18F-FDG PET/CT scan.The imaging results were compared with final clinical diagnosis.The diagnostic sensitivity,specificity,positive predictive value,negative predictive value and accuracy of 18F-FDG PET/CT were calculated.Two-sample t test was applied to compare lesion SUVmax of malignant and benign diseases.One-way analysis of variance was applied to compare SUVmax of highly,moderately and poorly differentiated tumors.x2 test was used to compare the difference of diagnostic sensitivities for hilar cholangiocarcinomas and common bile duct tumors.Results All patients were confirmed by exploratory laparotomy and subsequent histologic examination.A total of 94 cases (93 adenocarcinomas and 1 squamous carcinoma) were confirmed malignant and 22 cases (11 biliary calculi,9 cholangitis,1 choledochal cyst,1 tuberculosis) were confirmed benign.The diagnostic sensitivity,specificity,positive predictive value,negative predictive value and accuracy of 18F-FDG PET/CT for primary tumor were 61.7％ (58/94),77.3％ (17/22),92.1％ (58/63),32.1％ (17/53),64.7％ (75/116),respectively.The diagnostic sensitivity and specificity for regional lymph node metastasis were 45.5％(15/33),91.4％(53/58),and those for distant metastasis were 3/4,94.3％(82/87).The SUVmax of malignant tumors were higher than that of benign lesions (4.57± 3.75,2.72± 2.48;t =2.83,P＜ 0.05),while the differences of SUVmax among highly,moderately and poorly differentiated tumors were not significant (4.89±4.75,4.23±2.49,4.47±2.73;F=0.269,P＞0.05).18F-FDG PET/CT showed a lower sensitivity in hilar cholangiocarcinomas than that in common bile duct tumors,while no statistical significance was observed:48.6％ (17/35) vs 69.0％ (40/58),x2=3.827,P＞0.05.Conclusions The value of 18F-FDG PET/CT in preoperative diagnosis and staging of EHCC is limited.It can distinguish some benign diseases from malignant tumors,but with higher false positive for cholangitis.It can help to adjust treatment strategies by detecting distant metastasis.

Objective To investigate the consistency and stability of two types of pubococcygeal line (PCL) determined by dynamic MRI used in evaluating pelvic organ prolapse (anterior and apical compartments).The first type of PCL was measured from the inferior pubic symphysis to the tip of coccyx (PCLtip) and the second was to the sacrococcygeal joint (PCLjnt).Methods Dynamic MRI changes of 50 female patients who were diagnosed with pelvic organ prolapse by pelvic organ prolapse quantification were retrospective reviewed.Chi-square test was used to compare the staging of each pelvic compartment (anterior,apical) with the two PCLs.The lengths and the degree of the oblique angle of the two PCLs during the rest and straining were compared using a paried t test.Results Agreement of PCLjnt with PCLtip was 96％ (48/50) for anterior compartment and 94％ (47/50) for apical compartment.There was no difference between the two PCLs in staging of each pelvic compartment (anterior,apical)(x2 values were 2.000 and 3.000,P values were 0.368 and 0.223).The length of the PCLtip at rest and straining was (10.1±0.8),(10.2± 0.8) cm respectively and the result was statistical significance (t=-2.339,P=0.023).Twenty patients (40％) in the 50 pelvic organ prolapse patients demonstrated a shortening of the PCLtip,while the rest including 30 patients (60％) was longer.The oblique angle of the PCLtip at rest and straining was 22°±6° and 18°±11° respectively(t=3.490,P=0.001).The length of the PCLjnt at rest and straining were (11.2±0.8) and (11.2± 0.8)cm respectively(t=-1.845,P=0.071).The oblique angle of the PCLjnt at rest and straining were 29°±6° and 26°± 10° (t=2.836,P=0.007),but the degree of PCLjnt's oblique angle had a mild fluctuate compared with the PCLtip.Conclusions PCLjnt and PCLtip have the equal level in staging of anterior and apical pelvic organ prolapse.Meanwhile the oblique angle and the length of PCLjnt illustrated the better the stability.

Objective To investigate the influence on biological characteristics in human pancreatic cancer cells after beding transfected by two anti-carcinoma miRNAs at the same time.Methods Pancreatic cancer cells PANC1,SW1990 and normal pancreatic cells AR42J were transfected by miR-34a and(or) miR-let7 by liposome.Cells transfected with negative control miRNA (miR-NC) and untransfected were as controls.The expression of miR-34a and miR-let7 were detected by real-time fluorescent quantitative RT-PCR.The cell proliferation was detected by MTT test and the migration and invasion were evaluated by transwell assay.The apoptosis rate was measured by flow cytometric analysis.Results After being transfected with miRNAs,the expression of miR-34a and miR-let7 in double transfection group (miR-34a and miR-let7 were transfected at the same time),miR-34a transfection group,miR-let7 transfection group was significantly up-regulated than those in miRNA-NC transfection group and untransfected group in PANC1 cells,SW1990 cells and AR42J cells,repectively.The difference which was statistically significant (P ＜0.05)indicating that cells were successfully transfected.The cell proliferation in double transfection group of PANC1 cells and SW1990 cells were (0.665 ± 0.01,0.6375 ± 0.03),which were significantly inhibited compared with (0.974 ± 0.03,0.971 ±0.05) in miR-NC group and (0.8875 ±0.05,0.8625 ±0.06) in miR-let7 group.The difference was statistically significant(P ＜ 0.05).The cell proliferation activity in double transfection group was lower than those in miR-34a group (0.795 ±0.06,0.7925 ±0.06),but did not have statistically significant difference.There was no significant change in AR42J cells.Cell invasion assay showed that the number of PANC1 cells permeating substrate membrane in miR34a group (103.7 ± 3.28) and miR-let7 group (100.7 ± 1.76) were significantly fewer than miR-NC group (231.3 ±2.6) and untransfected group (153.7 ±2.6).The number of cells permeating substrate membrane in double transfection group(61.67 ± 3.18)was fewer than miR-34a group and miR-let7 group,respectively.The difference was statistically significant (P ＜ 0.01).The migration test had consistent results with invasion test.The changes of invasion and migration in SW1990 cells were similar to those in PANC1 cells.The apoptosis rate of PANC1 cells in miR-34a group,miR-let7 group,double transfection group,miR-NC group and untransfected group was (16.66 ± 1.27) ％,(15.46 ± 0.33) ％,(23.35 ± 1.80) ％,(9.33 ± 0.31) ％ and (8.83 ± 0.36) ％ respectively.Single transfection group had higher apoptosis rate than miR-NC group and untransfected group (P ＜0.05).Double transfection group had a significantly higher apoptosis rate than miR-let7 group (P ＜ 0.05),while there was no significant difference between double transfection group and miR-34a group.Conclusions The cell proliferation,invasion and migration in double miRNAs transfected pancreatic cancer cells were significantly down-regulated compared with those in single miRNA transfected cells,while apoptosis rate in double miRNAs transfection group was higher than single miRNA transfection group.Thus,the combination of two anti-cancer miRNAs may exert a more significant synergistic antitumor effect.

Female ; Humans ; Leukemia, Promyelocytic, Acute ; Pregnancy ; Tretinoin

OBJECTIVETo investigate the viability of tissue-engineered heart valve leaflets prepared with cell-polymer constructs in nude mice.

METHODSSheep endothelial cells and smooth muscle cells/fibroblasts were seeded on patches of PHA and implanted subcutaneously in athymic mice (BALB/C). The cell-polymer constructs were harvested 12, 14, 21 and 28 days after implantation.

RESULTSFourteen days after implantation, the cell-polymer constructs exhibited similar color with the autologous tissues, and HE staining showed more numerous cells in the implant. At 28 days following implantation, muscular fibers were formed in the cell-polymer constructs. V-G staining showed positive collagen staining in the implant at 12 days after implantation, while the control implants retrieved 28 days after implantation did not show extensive tissue formation or muscular fiber formation.

CONCLUSIONThe cell-polymer constructs can survive in vivo and has the potential to grow into autologous valve leaflets in the nude mice.

Animals ; Bioprosthesis ; Endothelium, Vascular ; cytology ; Heart Valves ; Implants, Experimental ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Muscle, Smooth, Vascular ; cytology ; Sheep ; Tissue Engineering ; methods

OBJECTIVETo present an improved method to obtain pure, viable, freshly isolated hepatic stellate cells.

METHODSAdult male SD rats were used. All procedures were performed with the animals under sodium pentobarbital anesthesia. Three days after the single intravenous administration of 1 ml liposome-encapsulated CL2MDP, which has selective cytotoxicity of Kupffer cells, livers were perfused with D-Hank's solution containing 100 U/ml heparin for 10 to 15 minutes, and then with 0.05% collagenase dissolved in D-Hank's solution for 25 to 30 minutes. The liver was then gently homogenized and further incubated in 0.025% collagenase, and 0.005% DNAase I for 30 minutes at 37 degrees C under constant stirring. This suspension was filtered through stainless steel gauze and centrifuged for 2 minutes at 50 x g to remove parenchymal cells. Sinusoidal cells in the supernatant were recovered by centrifugation for 10 minutes at 300 x g. The cells were resuspended in the presence of 28.7% Nycodenz stock solution. The final concentration of Nycodenz at this stage was 11.5%. Following centrifugation for 17 minutes at 1400 x g, The cells at the top of this Nycodenz solution were collected. Cells were resuspended in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum, The cells were seeded in 50 ml culture flask at a density of 500,000 cells/ml, The cell viability was determined by trypan blue exclusion staining, the purity of hepatic stellate cells was identified by the expression of Desmin using immunocytochemistry method. Endogenous peroxidase staining was used to detect Kupffer cells.

RESULTSThe yield rate of hepatic stellate cells was 3 x 10(7) per rat, the cell viability was more than 95%, the desmin positive cell rate was 90%, no endogenous peroxidase positive cells were detected.

CONCLUSIONThe method for the isolation of hepatic stellate cells was developed without Kupffer cells confusion. The availability of highly purified stellate cells will facilitate the investigation of their functions in primary culture.

Animals ; Cell Culture Techniques ; Cell Separation ; methods ; Kupffer Cells ; cytology ; Liver ; cytology ; Male ; Rats ; Rats, Sprague-Dawley ; Ultracentrifugation ; methods

Adolescent ; Antigens, Viral ; blood ; Child ; Child, Preschool ; Cytomegalovirus ; immunology ; Cytomegalovirus Infections ; diagnosis ; Female ; Flow Cytometry ; Hepatitis, Viral, Human ; diagnosis ; virology ; Humans ; Infant ; Male ; Phosphoproteins ; blood ; Viral Matrix Proteins ; blood