Neural stem cells were labeled with superparamagnetic iron oxide (SPIO) and tracked by MRI in vitro and in vivo after implantation. Rat neural stem cells were labeled with SPIO combined with PLL by the means of receptor-mediated endocytosis. Prussian blue staining and electron microscopy were conducted to identify the iron particles in these neural stem cells. SPIO-labeled cells were tracked by 4.7T MRI in vivo and in vitro after implantation. The subjects were divided into 5 groups, including 5 × 105 labeled cells cultured for one day after labeling, 5 × 105 same phase unlabeled cells, cell culture medium with 25 μg Fe/mL SPIO, cell culture medium without SPIO and distilled water. MRI scanning sequences included T1WI, T2WI and T2*WI. R2 and R2* of labeled cells were calculated. The results showed: (1) Neural stem cells could be labeled with SPIO and labeling efficiency was 100%. Prussian blue staining showed numerous blue-stained iron particles in the cytoplasm; (2) The average percentage change of signal intensity of labeled cells on T1WI in 4.7T MRI was 24.06%, T2WI 50.66% and T2*WI 53.70% respectively; (3) T2 of labeled cells and unlabeled cells in 4.7T MRI was 516 ms and 77 ms respectively, R2 was 1.94 s−1 and 12.98 s−1 respectively, and T2* was 109 ms and 22.9 ms, R2* was 9.17 s−1 and 43.67 s−1 respectively; (4) Remarkable low signal area on T2WI and T2*WI could exist for nearly 7 weeks and then disappeared gradually in the left brain transplanted with labeled cells, however no signal change in the right brain implanted with unlabeled cells. It was concluded that neural stem cells could be labeled effectively with SPIO. R2 and R2* of labeled cells were increased obviously. MRI can be used to track labeled cells in vitro and in vivo.
To investigate the high-risk factors for newborn hearing loss and to provide information for preventing the development of hearing loss and delaying its progression, from May 2003 to June 2006, neonates who failed to pass the universal newborn hearing screening (UNHS) were referred to Jinan Newborn Hearing Screening and Rehabilitation Center from 7 newborn hearing screening centers in seven cities of Shandong province. One-to-one pair-matched case-control method was employed for statistical analysis of the basic features of definitely identified cases. High-risk factors relating to the bilateral hearing loss were evaluated by univariate and multivariate Logistic regression analysis. Our results revealed that 721 transferred newborns who didn’t pass the hearing screening received audiological and medical evaluation and 367 were confirmed to have hearing loss. Of them, 177 neonates with hearing loss who met the matching requirements were included in the study as subjects. Univariate analysis showed that high-risk factors related to hearing loss incuded age of father, education backgrounds of parents, parity, birth weight, gestational weeks, craniofacial deformity, history of receiving treatment in neonatal intensive care unit (NICU), neonatal disease, family history of otopathy and family history of congenital hearing loss. Multivariate Logistic regression analysis revealed that 4 independent risk factors were related to bilateral hearing loss, including parity (OR=16.285, 95% CI 3.379–78.481), neonatal disease (OR=34.968, 95% CI 2.720–449.534), family history of congenital hearing loss (OR=69.488, 95% CI 4.417–1093.300) and birth weight (OR=0.241, 95% CI 0.090–0.648). It is concluded that parity, neonatal disease and family history of hearing loss are the promoting factors of bilateral hearing loss in neonates and appropriate intervention measures should be taken to deal with the risk factors.
In order to analyze the in vivo expression of Candida albicans secreted aspartyl proteinases (SAP) in human vaginal infection, the vaginal secretion from 29 human subjects was collected by vaginal swab, and the expression of SAP1-SAP6 was detected by reverse-transcriptase polymerase chain reaction using specific primer sets. It was found that Sap2 and Sap5 were the most common genes expressed during infection; Sap3 and Sap4 were detected in all subjects and all 6 SAP genes were simultaneously expressed in some patients with vaginal candidiasis. It was suggested that the SAP family is expressed by Candida albicans during infection in human and that Candida albicans infection is associated with the differential expression of individual SAP genes which may be involved in the pathogenesis of vaginal candidasis.
HOXA10 gene plays an essential role in differentiation of the endometrium and in human reproduction. The aim of this study was to investigate the regulatory effect of sex steroids and HB-EGF on HOXA10 gene in Ishikawa cells. Ishikawa cells were incubated with 17-beta estradiol (10−8 mol/L), medroxyprogesterone acetate (MPA) (10−6 mol/L), RU486 (10−5 mol/L) or HB-EGF (10 ng/mL) for 48 h respectively. The expression of HOXA10 gene was detected by immunofluorescence, reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. Our results showed that either estrogen alone, progestin alone or progestin combined with estrogen could significantly increase the expression of HOXA10 gene 48 h after the treatment (P<0.05). But estrogen combined with progestin and RU486 could inhibit the up-regulation by estrogen and progestin. HB-EGF could elevate the expression of HOXA10 gene 48 h after the treatment (P<0.05). It is concluded that both estrogen and progestin can up-regulate the expression of HOXA10 gene in Ishikawa cells, but RU486 can inhibit the effect and HB-EGF can elevate the expression level of HOXA10 gene.
The diagnosis and treatment of the lacrimal passage obstruction with lacrimal endoscope was investigated and its subsidiary surgical procedures were evaluated. Ninety-three patients (109 eyes) with lacrimal passage obstruction, including presaccal canalicular obstruction (PSCO) and nasolacrimal duct obstruction (NLDO), were examined under a lacrimal endoscope, and the obstructions were treated with laser or micro-drill. All patients were followed up after the operation for 3–6 months. The difference between the laser and the micro-drill treatment was observed. During the period of follow-up, the curative rate was 82.57%. The healing rate in PSCO group and NLDO was 80.36% and 84.91% respectively (P>0.05). After treatment with the laser, the healing rate was 93.33% in the PSCO group and 66.67% in the NLDO group respectively (P<0.05). After treatment with the micro-drill, the healing rate in PSCO and NLDO groups was 65.39% and 94.28% respectively (P<0.01). The lacrimal passage obstruction can be observed and treated directly through the lacrimal endoscope. Choosing different surgical procedures in operation according to the locations of the obstruction is helpful to improve the effectiveness.
To establish an animal model of benign lymphangiomas of C57BL/6 mouse in vitro and to observe the effect of mouse ascites melanoma cell B16-F1 on the lymphangiogenesis, 16 C57BL/6 mice aged 8 weeks were given two intraperitoneal injections of incomplete Freund’s adjuvant at a 15-day interval. The induced neoplasms were studied histopathologically and thhe neoplasms specimens were immunohistochemically examined for the expressions of VEGF-C (vascular endothelial growth factor-C) and Flt-4 (VEGFR-3, vascular endothelial growth factor receptor-3). The neoplasms were harvested and embedded in fibrin gel for culture in conditioned medium of B16-F1 cells in vitro and observed under inverted microscope. Our results showed that white solid tumor masses developed in peritoneal cavity after the induction. The tumors were confirmed to be lymphangioma by gross and histological examination. The tumor cells expressed both VEGF-C and Flt-4. Lymphatic capillaries coming from lymphangioma specimen grew into the gel and the conditioned medium of B16-F1 cells was found to be able to promote the growth of the vessels. It is concluded that intraperitoneal injection of incomplete Freund’s adjuvant is a good method for inducing benign lymphangiomas in mouse and B16-F1 cells can promote lymphangiogenesis.