Cholangiopancreatography, Endoscopic Retrograde ; Cholangiopancreatography, Magnetic Resonance ; Endosonography ; Humans ; Lymph Node Excision ; Neoplasm Staging ; Pancreatectomy ; methods ; Pancreatic Neoplasms ; diagnosis ; pathology ; surgery ; Tomography, X-Ray Computed ; methods

Peking University Shenzhen Hospital launched a reform to outsource its logistic support service to overcome the setbacks of overstaffing,poor service and high costs with the existing logistic departments since its opening.Practice of a decade has raised the quality of its logistic support system and lowered service costs.Furthermore,the reform has optimized the hospital's human resource structure,and improved performance of the hospital staff,paving the way for better social benefits of the hospital.

Objective To analyze the histopathology of patients with atypical squamous cells of undetermined significance (ASCUS),and provide evidence for further classification and clinical treatment of ASCUS.Methods The histopathology of cervical biopsy specimens from 249 patients with ASCUS diagnosed by liquid-based cervical cytology examination was analyzed retrospectively. Results Among the 249 ASCUS patients, the proportion of patients with inflammation was 34.14% (85/249), morphological change by human papillomaviral infection was 19.28%(48/249), CIN I was 32.53%(81/249),CIN II was 8.84%(22/249),CIN III was 3.21%(8/249), infiltrating squamous cell carcinoma was 1.20%(3/249),and endometrioid adenocarcinoma was 0.80%(2/249) . Conclusion It is very important to to further definitude the diagnosis of ASCUS, because a certain proportion of cervical cancer and precancerous leisions woud be confirmed.

Objective:To establish a simple HPLC method for the determination of warfarin in human plasma and analyze the re-sults of 100 monitoring reports. Methods:Warfarin was extracted from the plasma samples by acetonitrile. The extract was separated on a ZORBAX Eclipse XDB-C8 column (150 mm × 4. 6 mm, 5 μm)with a mobile phase consisting of methanol and 0. 1% acetic acid (65∶35) at flow rate of 1. 0 ml·min-1 and detected by a diode array detector. The detection wavelength was at 308nm and the sample size was 20 μl. Results:The calibration curve was linear within the range of 0. 1-5. 12μg·ml-1 . The average recovery and RSD was ranged from 85. 0% to 102. 3% and 1. 2% to 3. 9%(n=6), respectively. Inter-and intra-day RSD were 1. 1%-1. 8% and 1. 6%-2. 9%, respectively. Conclusion:The proposed method is rapid, convenient and accurate in the detection of warfarinin concentration in plasma.

OBJECTIVE:To study the in vitro dissolution of Enalapril maleate and folic acid tablet. METHODS:HPLC was performed on the column of Agilent HC-C18 with mobile phase A of acetonitrle-phosphate buffer solution(70:30,V/V) and mobile phase B of acetonitrle-phosphate buffer solution(5:95,V/V)(gradient elution) at a flow rate of 1.0 ml/min,detection wavelength was 215 nm,column temperature was 50 ℃,and volume injection was 80 μl. Media were water,hydrochloric acid solution(pH 1.2),phosphate buffer solution(pH 5.0)and phosphate buffer solution(pH 6.8),medium volume was 900 ml and rotation speed was 50 r/min. The dissolution behavior of enalapril maleate in Enalapril maleate and folic acid tablet in 4 media were studied and compared with the dissolution behavior in vitro in original preparation of Enalapril maleate tablet,meanwhile,the dissolution behar-ior of folic acid in Enalapril maleate and folic acid tablet in phosphate buffer solution(pH 5.0)were studied and compared with dis-solution data of folic acid preparation in Japanese Orange Book to evaluate the intrinsic quality. RESULTS:The linear range was 0.561-14.03μg/ml for enalapril maleate(r=0.999 9)and 0.043-1.085μg/ml for folic acid(r=0.999 9),respectively;RSDs of pre-cision and stability tests were lower than 2.0%;recoveries of enalapril maleate in 4 media were 100.63%-102.33%(RSD=0.72%, n=9),99.27%-100.44%(RSD=0.41%,n=9),99.71%-100.29%(RSD=0.15%,n=9)and 96.74%-99.19%(RSD=0.79%,n=9),respectively. Recoveries of folic acid were 100.18%-101.63%(RSD=0.48%,n=9),97.73%-101.81%(RSD=1.32%,n=9),99.60%-102.24%(RSD=0.74%,n=9)and 100.00%-102.76%(RSD=0.90%,n=9),respectively. In 15 min,the dissolution of enalapril maleate of 2 preparations in 4 dissolution media were more than 85%;dissolution speed of folic acid in Enalapril male-ate and folic acid tablet was faster than that in folic acid preparation in phosphate buffer solution(pH 5.0). CONCLUSIONS:The method is suitable to determine the dissolution of Enalapril maleate and folic acid tablet;the in vitro dissolution curve of enalapril maleate in Enalapril maleate and folic acid tablet is similar to Renitec,and the in vitro dissolution of folic acid is better than folic acid preparation.

Objective To explore the influencing factors for the purity and viability of primary cultured cortical neurons of rat,and optimize the separating and cultivating conditions of the cortical neurons.Methods The primary cotical neurons were cultured in a serum-free culture system of B27-supplemented neurobasal medium.The differences in purity and viability of primary cultured neurons between embr-yonic rat group and newborn rat [postnatal 24 h and 5 d] group were evaluated by morphology,immunocytochemistry of neuron-specific enolase(NSE) and trypan blue staining.The changes of neurons purity and viability in different trypsin digestion time(0 min,5 min and 15 min) at different environment temperatures(20 ℃ and 30 ℃) were assessed by immunocytochemistry and trypan blue staining.Results The primary cultured neurons from fetal and newborn rats grew well.There was no significant difference in embryonic rat and postnatal 1 d newborn rat group[(91.30?1.03)%,(89.50?1.78)% respectively in purity;and(98.20?0.58)%,(97.10?0.98)% respectively in viability].The neurons from 5-days newborn rat were inferior to that from fetal and 1-day newborn rat in purity and viability[(82.00?1.25)% and(92.87?1.56)% respectively].Shortening operation time and adjusting digestion time according to the environment temperatures could improve neuronal viability:A digestion for 15min at the environment temperature of 20 ℃ or for 5 min at 30 ℃ could acquired cells with higher viability [(98.20?0.58)% and(96.70?0.64)% respectively].Conclusions It is an easy,practical choice to culture priamry cortical neurons from postnatal 1 d newborn rats.Optimizing the separating and cultivating condition(environment temperatures,digest time,et al.) will improve the neurons purity and viability.

AIM:To investigate the effects of estrogen on the expression of matrix metalloproteinases-2(MMP-2), tissue inhibitor of metalloproteinases-2(TIMP-2) and transforming growth factor-β1(TGF-β1) in cultured human corneal stromal cells.METHODS: Inflammatory environments of human corneal stromal cells were simulated by using 1.5ng/mL IL-1β.The cells were then treated with or without different concentrations of estrogen(0, 1×10-4, 1×10-6, 1×10-8, 1×10-10mol/L estradiol)in vitro.Cell viability was evaluated by MTT.Expression levels of MMP-2, TIMP-2 and TGF-β1 proteins were measured by enzyme-linked immunosorbent assay(ELISA).RESULTS:Estrogen did not affect the viability of human corneal stromal cells.Compared with the control group, expression levels of MMP-2 and TGF-β1 proteins in E2 treatment group significantly decreased after being treated with estrogen, while the expression level of TIMP-2 significantly increased.CONCLUSION: Estrogen could, to some extent, down-regulate the expression of MMP-2 and TGF-β1 and up-regulate the expression of TIMP-2, which might contribute to protecting human cornea.

Objective To investigate the effect of transplantation of bone marrow mesenchymal stem cells (MSCs) in subarachnoid space on chronic neuropathic pain in rats. Methods One hundred and forty female SD rats weighing 180-200 g were used in this study. Chronic neuropathic pain (NP) was induced by ligation and separation of tibial and common fibular nerves (SNI). Two weeks after the surgery the animals were randomly divided into4 groups (n=35 each):group Ⅰ NP; group Ⅱ NP+ MSC (MSCs); group Ⅲ NP+ phosphate buffer saline (PBS) and group Ⅳ NP+ bone marrow monocyte (BNMCs). MSCs 10 μl, PBS 10 μl and BNMCs 10 μl were injected into subaraclmoid space at 2 weeks after surgery in group Ⅱ , Ⅲ and Ⅳ respectively. Paw withdrawal threshold to yon Frey filament stimulation (PWT) was measured before surgery (T0, baseline), at 2 weeks after surgery (T1) and 1, 2, 3, 4 and 5 weeks after subarachnoid injection (T2-6). Five animals were killed at T1-6 in each group and their lumbar enlargements were removed for determination of BDNF mRNA expression. Results PWT was significantly decreased by SNI at T1 in all 4 groups, and at T2-6 in group Ⅰ , Ⅲ and Ⅳ as compared with the baseline at T0 (P ＜ 0.05). Subarachnoid MSC transplantation significantly reduced mechanical hyperalgesia at T2-6 and up-regulated BDNF mRNA expression at T2-4 as compared with that at T1 (P ＜0.05). There was no significant change in PWT and BDNF mRNA expression after subarachnoid PBS and BNMCs injection in group Ⅲ and Ⅳ (P ＞ 0.05). Compared with group Ⅰ , PWT was significantly increased and BDNF mRNA expression was up-regulated in group Ⅱ (P ＜ 0.05), but no significant change in PBS and BNMCs was found in group Ⅲ (P ＞ 0.05) .Conclusion Up-regulation of BDNF mRNA expression in the spinal cord may be involved in the amelioration of chronic neuropathic pain by subarachnoid bone marrow mesenchymal stem cell transplantation.

BackgroundPterygium is an ocular surface disease of abnormal cell proliferative kind and angiogenesis plays an important role in its development and recurrence.Several anti-angiogenic therapies have been used to treat pterygium,but there very few studtes for the in vivo observation of the microvessles in pterygium.ObjectiveThis study was to observe angiogenesis in pterygium with a high-resolution confocal microscope in vivo and to perform immunohistochemical study in vitro.MethodsA prospective case-controlled study was designed.Twenty eyes of 20 consecutive patients with primary pterygia and 20 age- and sex-matched patients with inner eye diseases and strabismus with normal conjunctiva were enrolled in this study.An in vivo confocal microscopy imaging system (Heidelberg Retina Tomograph Ⅱ Rostock Cornea Module) was used to collect microvascular pictures from the anterior part of pterygia and normal nasal conjunctiva of controls,and then immunochemistry was performed to examine the expression of CD31 in microvessel in vitro.The vascular density values were compared between these two groups.The correlation of vascular density values between in vivo Heidelberg Retina Tomograph and in vitro immunohistochemistry was calculated.Written informed consent was obtained from pationts before any examination and surgery.ResultsUnder the in vivo confocal microscope,the microvessel density was (8929±2993) μm/mm2 and (4202 ±692)μm/mm2,respectively,in pterygium and the normal conjunctiva group with a statistically significant difference between them (t =6.881,P＜0.01 ).Immunochemistry revealed that the expression of CD31 to measure vascular density was ( 21.00 ± 4.06/400 × field ) and ( 6.07 ± 1.75/400 × field ) in pterygium and the normal conjunctiva group,showing significant difference (t =12.312,P＜0.01 ).Positive correlations were found in the vascular density values between in vivo corneal laser confocal microscopy examination and in vitro immunochemistry examination in both the pterygium group and normal conjunctiva group (pterygium group:r=0.649,P＜0.01 ;normal conjunctiva group: r=0.572,P＜0.01 ) ConclusionsIn vivo confocal microscopy imaging is superior to in vitro immunochemistry in evaluating the microvessel of pterygium.The results of this study offer a new way index for further investigation of the biological behavior of pterygium and its mechanism.

Objective To explore human umbilical cord blood mesenchymal stem cells(MSCs)colonization in hypoxic-ischemic encephalopathy(HIE)rats brain.Methods Models of 7-day-old newborn rats with HIE brain injury were established.Meanwhile,on the same day,MSCs were transplanted with Hoechst 33258 for 24 hours into rats models marked by flurescent nucleotide dye injected through caudal vein or with stereotactic instrument.After 15-30 days,then MSCs were detected with fluorescene microscope.Results With the improved rice methods,HIE animal model was successfully attained.Majority of MSCs were distributed in the cortex,hippocampus of the lesioned hemisphere,especially in the forehead.And abtained a good fusion with HIE rats brain tissue.Conclusion Human umbilical cord blood MSCs can be cultured,when transplanted into the HIE rats model,they can move into intracranial,and integer with rats brain tissue.