Objective To isolate and identify the photoreceptor precursor cells isolated from newborn mouse retina. Methods Fourteen newborn littermate mice were randomly divided into seven groups (P1-P7,2 each). The retinal cells were isolated from the newborn mice on postnatal 1-7d respectively,and then cultured in vitro. The cellular growth state was observed under inverted phase contrast microscope. After in vitro expansion,5-bromo-2-deoxy-uridine (BrdU),neuroepithelial stem protein (Nestin),neural retina leucine zipper (Nrl) and Opsin in cultured cells were detected by immunohistochemistry and immunofluorescence techniques. Results Most of retinal cells isolated from the seven groups (P1-P7) of newborn mice proliferated and adhered to the plate in the particular medium. Most of the growing cells were BrdU-positive cells. A part of the cells could express neural stem cell specific antigen-Nestin,the positive expression rate of Nestin in the seven groups (P1-P7) were 31.0%,31.6%,32.3%,30.2%,31.2%,30.9% and 29.5%,respectively,with no significant difference. Some growing cells expressed Nrl,and the expression of Nrl was increased significantly on the third day after birth,and then increased in small range,the positive expression rate of Nrl in the seven groups (P1-P7) were 20.6%,35.2%,65.5%,68.6%,71.6%,73.0% and 73.3%,respectively. The positive expression of Opsin was found only in a few cells of P5-P7,while the cells in groups P1-P4 did not express Opsin. Conclusions The photoreceptor precursor cells are presented in neonatal mouse retina. They have the ability of proliferation and may differentiate into retinal photoreceptor cells,having a potentiality of expressing Opsin. The photoreceptor precursor cells are increased significantly on the third day after birth,and then differentiate into mature retinal photoreceptor cells gradually.

Objiective To observe the morphological changes of rabbits' retina at different time points at which seawater was injected into the vitreous of rabbits. Methods Twenty-five New-Zealand white rabbits were randomly assigned to five groups, as 30 minutes, 2 hours, 6 hours, 12 hours and 24 hours group following the seawater injection in the vitreous. The right eyeball served as experimental eyeball and the left one as control in each group. 0.5ml artificial seawater was injected into the intermedius of rabbits right vitreous. 0.5ml balanced salt sdution was injected into the left vitreous in teh same way. At different time points following the eyeball injury, the morphological changes of retina were observed with light microscope and transmission electron microscope. Results The retina of control eyeballs had clear layers and regular cell shape in retinal structures with conventional staining. For the experimental eyeballs, the ganglion cell layer began to swelling after 30 minutes of seawater injection. The nerve fiber swelled obviously and the mitochondria vesiculated and swelled in the inner segment at 2 hours of seawater injection. The cell population of the inner nuclear layer began to reduce in 6 hours. At 12 hours of seawater injection, the retinal damages included disorganized cell nuclei, decreased cell population in ganglion cells and inner nuclear layer. Mitochondria vesiculated in photoreceptor cells, nuclear membrane shrank and sank inward. The vesiculation on inner and outer segments increased obviously. Conclusions Seawater may cause the morphological changes of rabbits' retina at early stage, and the histological damages become aggravation following the time prolongation. The damages would be unrecoverable 6 to 12 hours after seawater injury.

ObjectiveTo evaluate the past 5 years' clinic experience of diagnosis and treatment of the pediatric soft-tissue foreign body (STFB),and to probe new strategies for its clinical management.MethodsTotally 165 consecutive children with small radiopaque STFB were involved.All the children were diagnosed with X-rays.CT (enhanced CT in 9 children)and virtual anatomy imaging (VAI) were performed in 40 children.Percutaneous foreign body forceps removal guided with C-arm video-fluoroscopy was performed and the effect was evaluated.ResultsThe longest dimension of STFB ranged from 1 mm to 40 mm,and the shortest dimension ranged from 1 mm to 5 mm,including scrap-iron,broken iron nails and needles,and glass pieces embedded in soft tissues under surface of the limbs,neck,chest,abdomen and pelvis.Seventy-six (76/165,46.06 ％) children received interventional therapy,and 73 were completely cured (73/76,96.05 ％),2 were partially cured (2/76,2.63 ％),and 1 was failed (1/76,1.32 ％).VAI accurately depicted STFB closely to large vessels even associated vascular complication with local large hematomas or pseudoaneurysms,helped to select the treatment methods and the forceps removal roads.Hematoma,infection,neural damages and other serious complications did not occur during and after operation.ConclusionVideo-fluoroscopy-guided percutaneous foreign body forceps removal is minimally invasive,safe and effective for small radiopaque STFBs,but may not suitable for the one very close to large blood vessels with or without vascular injuries complications.Preoperative CT VAI is helpful to locate STFB within complicated anatomic structures,selecting optimal intervention pathway and assessing the risk of intervention.

Objective To explore the clinical value and the difference of bone mineral density(BMD) in every vertebra of lumbar spine in middle old-aged Chinese female with quantitative computed tomography (QCT).Methods The BMD in every vertebra of lumbar spine was investigated by QCT in 725 healthy females aged from 35 to 98 years. They were divided into groups every five years.The BMD in vertebra of lumbar spine (L 1～L 4) were measured by QCT and the groups tested for significant differences,using SPSS10.0 software for the analysis.Results Lumbar spine (L 1～L 4) BMD declined with aging. The bone quantity was predominantly declined in group of 50 to 54 years (?

Objective To establish a stable animal model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were replicated. After immediate subcutaneous suture,the mice were randomly divided into four groups,ie,Group A was inoculated with 50 μl sterile PBS solution),Groups B,C and D were inoculated with 50 μl suspension containing 1 × 106,1 × 108 and 1 × 1010 colony forming unit (CFU)/ml bioluminescent methicillin-resistant staphylococcus aureus (MRSA) respectively.Then,the diet behavior of each group was observed and the mean weight and mortality of each group were also recorded at different time points.The bioluminescent intensity of fluoresce in the wounds was recorded at different time points by using the charge-coupled device (CCD) based imaging system.Local wound tissues were incised at 24 hours after inoculation for HE staining so as to observe wound inflammatory reaction in each group.Wound healing time of each group was also recorded. Results ( 1 ) Average weight:Groups A and B showed unobvious changes in weight; Group C lightened until day 3 after inoculation and then recovered gradually to the preinoculation level at day 14; Group D lightened gradually until death.(2)Mortality:Groups A and B had no death; Group C had 10％ deaths at day 14; Group D had 100％ deaths.(3) Bioluminescent intensity of wounds:Groups A and B showed a gradual weakened luminescence since the day of inoculation and had almost complete disappearance at days 5 and 7 respectively; there was no sign of obvious increase or decrease in Group C from the day of inoculation till day 14 ; Group D had a gradual increase since the day of inoculation and the luminous area expanded until the death.(4) HE staining at 24 hours after inoculation:all the four groups showed inflammatory cell infiltration,especially in Groups C and D.(5) Wound healing time:wound healed at days 5 and 7 after inoculation in Groups A and B; the wounds showed no healing even at day 14 in the Group C,but the wounds length and area did not show obvious enlargement or diminishment ; the wounds extended gradually until the death in the Group D,since the day of inoculation. Conclusions The inoculation of 50 μl suspension with 1 × 108 CFU/ml bioluminescent MRSA to full-thickness skin incision rats allows direct,real-time dynamic and continuous detection of the occurrence and development of the wound infections.The infection model is easy to make and has stability and high repeatability.

OBJECTIVE:To optimize the extraction technology of asiaticosides from Centella asiatica. METHODS:Using to-tal amounts of asiaticoside and hydroxy asiaticoside as investigation indexes,single factor and orthogonal test were used to investi-gate the enzyme amount,enzymolysis time,enzymolysis temperature,ethanol volume fraction,liquid material ratio and ultrasonic extraction time and optimize the extraction technology of asiaticosides from C. asiatica by ultrasonic-enzyme method,and verifica-tion test was conducted. RESULTS:Optimal extraction technology was as follow as cellulose dosage of 12 mg/g,10-fold liquid ma-terial ratio added into 60% ethanol,enzyme hydrolysis for 60 min at 60 ℃,ultrasonic assisted extraction for 50 min. Average ex-traction rate of total asiaticosides was 1.92%(RSD=1.83%,n=3)in verification test. CONCLUSIONS:Ultrasonic-enzyme meth-od is stable and feasible for the extraction technology of asiaticosides from C. asiatica.

Objective To evaluate the effect of endoplasmic reticulum stress in trophocytes,in patients with intrahepatic cholestasis of pregnancy (ICP).Methods Sixty-one pregnant women who were hospitalized in Women's Hospital,School of Medicine,Zhejiang University from January to December 2015 were recruited.Thirty-one women who were diagnosed as ICP were defined as the ICP group and 30 healthy pregnant women were defined as the control group.The localization and expression intensity of glucose regulated protein 78 (GRP-78) in placental tissues were detected by immunohistochemistry technique.Electronic microscope was used to observe ultra-microstructure change of the endoplasmic reticulum in trophocytes and cell line Swan71.Reverse transcription (RT)-PCR and western blot were used to investigate the expression of GRP-78 mRNA and protein in Swan 71 cell.Results (1) GRP-78 protein was mainly expressed in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts.The protein expression of GRP-78 in placentas of the ICP group (13.2±2.4) was significantly higher than that in the control group (7.8±1.3,P＜0.01).(2) The volume of endoplasmie reticulum did not increase and the microvilli developed well,with no swelling and no expansion of endoplasmic reticulum in the control group.In the ICP group,microvilli injury,endoplasmic reticulum edema were found;the volume of endoplasmic reticulum increased,with dilation,vacuolation and significant degranulation.After treated with 100 μmol/L cholyglycine for 24 hours,universal dilatation of the endoplasmic reticulum were seen in the Swan71 cells.(3) In Swan71 cells,cholylglycine displayed a concentration-dependent up-regulation on the expression of GRP-78.The expressions of GRP-78 mRNA in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.17,2.17±0.16,5.47±0.36,5.65 ± 0.82,respectively.The expression of GRP-78 protein in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.04,1.17±0.15,1.33±0.13,1.73±0.13,respectively.The expression of GRP-78 mRNA and protein in 100 and 50 μ mol/L cholylglycine experimental group were significantly higher than 0 μmol/L (all P＜0.01).Conclusion The obvious expansion of endoplasmic reticulum and the increased expression of GRP-78 in trophocytes indicated that endoplasmic reticulum stress of trophocytes may be involved in the pathogenesis of ICP.

Objective: To explore the clinical effects of superior gluteal artery perforator island flap in repair of sacral pressure ulcer. Methods: From May 2012 to May 2017, 20 patients with sacral pressure ulcers (14 males and 6 females, aged 27 to 67 years) were admitted to our department. According to the consensus staging system of National Pressure Ulcer Advisory Panel in 2016, 6 cases were in 3 stages, 14 cases were in 4 stages, with the area of pressure ulcers ranging from 5.0 cm×4.0 cm to 10.0 cm×8.0 cm. After debridement and vacuum sealing drainage, the superior gluteal artery perforator island flaps were used to repair the pressure wounds, with the area of flaps ranging from 6 cm×5 cm to 13 cm×8 cm. The donor sites were sutured directly. The survival of flaps after operation, the healing of wounds, and the follow-up of patients were observed. Results: After surgery, flaps of 20 patients survived well without reoperation. The length of hospital stay of patients was 20 to 40 days, with an average of 25 days. Eighteen patients were followed up for 6 to 24 months, with an average of 12.2 months. The flaps were in good shape and elastic recovery. There were no complications such as seroma or hematoma in the donor sites. Both the patients and family members expressed satisfaction with the shape and texture of the flap and shape of hip. Conclusions The superior gluteal artery perforator island flap is reliable in blood supply and easy to rotate. The flap can carry a little muscle to increase the anti-infective ability. Moreover, the donor site can be directly sutured with slight damage. Thus, it is one of the good methods for repairing sacral pressure ulcers.

Objective To compare the clinical characteristics and influence factors of hyperuricemia between genders,in order to provide references for better controlling and preventing the occurrence and development of hyperuricemiaprovide.Methods A total of 5 783 people who underwent physical examination in two Baoding Health Screening Centers from January 1st to June 1st,2016 were enrolled in this study,and all volunteers completed physical examination,laboratory examination and questionnaire survey.Patients with hyperuricemia were selected to analyse the clinical characteristics and influence factors.Results There were statistically significant differences in clinical characteristics,including obesity,blood pressure,blood glucose,blood lipids,liver function,renal function,anemia,blood rheology examination,thyroid ultrasound and lateral radiographs,between male and female patients with hyperuricemia (P＜0.05).The survey showed that there were statistically significant differences in age,education level,marital status,work status,sleeping status,smoking and drinking between male and female patients with hyperuricemia (P＜0.05).The red blood cells counts,marital status and education level were influence factors for female patients with hyperuricemia,while have little effect on male patients.The smoking,creatinine and diastolic blood pressure were influence factors for male patients with hyperuricemia,while have no effect on female patients.Conclusion The clinical characteristics and the influencing factors of male and female patients with hyperuricemia are different,so corresponding preventive and therapeutic measures should be taken for male and female patients.

Despite recent studies, relatively few are known about the diversity of fungal communities in the deep Atlantic Ocean. In this study, we investigated the diversity of fungal communities in 15 different deep-sea sediments from the South Atlantic Ocean with a culturedependent approach followed by phylogenetic analysis of ITS sequences. A total of 29fungal strains were isolated from the 15 deep-sea sediments. These strains belong to four fungal genera, including Aspergillus, Cladosporium, Penicillium, and Alternaria. Penicillium, accounting for 44.8% of the total fungal isolates, was a dominant genus. The antiaflatoxigenic activity of these deep-sea fungal isolates was studied. Surprisingly, most of the strains showed moderate to strong antiaflatoxigenic activity. Four isolates, belonging to species of Penicillium polonicum, Penicillium chrysogenum, Aspergillus versicolor, and Cladosporium cladosporioides, could completely inhibit not only the mycelial growth of Aspergillus parasiticus mutant strain NFRI-95, but also the aflatoxin production. To our knowledge, this is the first report to investigate the antiaflatoxigenic activity of culturable deep-sea fungi. Our results provide new insights into the community composition of fungi in the deep South Atlantic Ocean. The high proportion of strains that displayed antiaflatoxigenic activity demonstrates that deep-sea fungi from the Atlantic Ocean are valuable resources for mining bioactive compounds.