Objective To investigate the effects of leflunomide(A77 1726) on the proliferation and apoptosis of human keratinocytes. Methods Proliferating cell nuclear antigen (PCNA) of HaCaT cells was analyzed with crystal violet staining and immunohistochemistry. The moiphological change was assessed with hematoxylin and eosin staining. The changes of cell cycle and apoptosis rate were measured by flow cytome-try. Result Epidermal proliferation was inhibited by leflunomide, at concentration 5 ?mol/L or more, which was begun after 24 hrs and manifested by PCNA positive cell decreasing. With the increasing of time or dosage, the anti- proliferation effect of leflunomide was significant. Meanwhile, the PCNA positive cell de-creased respectively. There was no PCNA positive cell while the drug concentration achieved 200 ?mol/L. Therefore, leflunomide significantly inhibited the proliferation of HaCaT cells in a dose-dependent and time-dependent manner. The morphological change and cell cycle change was found but no change of apoptosis rate was measured. Conclusion Leflunmide can inhibit the proliferation of HaCaT cell, without the effect on its apoptosis.

In order to improve the result of clinical practice for undergraduate students of sto-matology, the standardized training program was carried out before clinical practice by the teaching and researching Office of Periodontology of Affiliated Stomatological Hospital of Chongqing Medical University, in this program, based on our own realities, the structure and content of teaching was designed to guide the transformation of students to doctors, collective teaching and personalized teach-ing was combined to reform the conventional model of teaching and various student-oriented teaching methods were explored, including mimic forum of disciplines, intractable case discussion, clinical practice summary, course completion test and so on, thus promoting the result of clinical practice. Be-sides, the construction of teaching staff was strengthened, and the evaluation and management system was established to ensure the teaching quality, thus summarizing a set of efficient and effective clinical teaching methods presented in this paper.

OBJECTIVETo establish a new method and validate its feasibilities for quality evaluation of Fructus Epimedii.

METHODFour main effective flavones, epimedin A, epimedin B, epimedin C and icariin were selected as analytes to evaluate the quality of Fructus Epimedii. The relative correction factors (RCF) of icariin to the other three flavones were calculated. The method was evaluated by comparison of the quantitative results between external standard method and QAMS method.

RESULTNo significant differences were found in the quantitative results of three flavones by external standard method and QAMS method.

CONCLUSIONIt is feasible and accurate to evaluate the quality of Fructus Epimedii.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; Reproducibility of Results ; Statistics as Topic ; methods ; Time Factors

BACKGROUND: Recently biodegradable hydrogel has been extensively used to delivery anticancer drug and bioactive macromolecule. However, to protect the activity of the bioactive macromolecule, we need to obtain series of hydrogel which have milder hydrogelation conditions and shorter hydroglation time.OBJECTIVE: To prepare enantiomeric poly(L-Lactic acid) (PLLA)-poly(ethylene glycol (PEG)-PLLA/ poly(D-Lactic acid) (PDLA)-PEG-PDLA stereocomplex hydrogel which has shorter hydroglation time, to physically encapsulate a model drug-lysozyme and sustained release it from the hydrogel. METHODS: Triblock copolymers of PLLA-PEG-PLLA and PDLA-PEG-PDLA were synthesized by ring-opening polymerization of L(D)-lactide using PEG as the initiator and Sn(Oct)2 as the catalyst. The triblock copolymers were characterized by 1H nuclear magnetic resonance, FT-IR and X-Ray diffractometry. A hydrogel was prepared from an aqueous mixture of PLLA20-PEG227-PLLA20 and PDLA21-PEG227-PDLA21 (10 wt% concentration) at room temperature for 12 hours. X-Ray diffractometry test was used to research the gelation mechanism. The release profile of the lysozyme as a model drug from the hydrogel was tested. The morphology of the freeze-dried hydrogel was investigated by scanning electron microscope. The cytotoxicity of the hydrogel was evaluated by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide) assay.RESULTS AND CONCLUSION: Triblock copolymers of PLLA-PEG-PLLA and PDLA-PEG-PDLA were obtained. Both the PEG and PLA blocks in the copolymers could crystallize, but the crystallization of the PEG block was predominant. The stereocomplex formation between the PLLA and PDLA blocks within the hydrogel was confirmed by the X-Ray diffractometry analysis. The release profile of the lysozyme from the hydrogel exhibited a sustained-release pattern with a duration period of 7 days. The hydrogel exhibited a 3D interconnected porous structure with 50-100 μm pore size after being freeze-dried. The mouse fibroblast cell viability percentage was 99.3% after the cells contacted with the 100% extracted liquid for 72 hours.

Background It is thought in recently that photodynamic therapy (PDT) is an effective treatment method for chronic central serous chorioretinopathy (CSC), but the dosage of verteporfin and its long-term efficacy and complications is rarely elucidated ever before.Objective This study was to observe the long-term efficacy and safety of 60％ dose verteporfin PDT for chronic CSC.Methods This is a retrospective study and a self-controlled design was used.The clinical data of 25 eyes of 21 chronic CSC patients who received 60％-dose verteporfin PDT in Henan Eye Institute from January 2009 to May 2010 were reviewed, with the male 18 (85.71％) and female 3 (14.29％) as well as monocular CSC 17 patients and binocular CSC 4 patients.The average ages of the patients were (43±5) years.Fundus fluorescein angiography (FFA) , indocyanine green angiography (ICGA), optical coherence tomography(OCT) and best corrected visual acuity (BCVA) were examined in all the patients before and after treatment.PDT with the 60％-dose verteporfin (3.6 mg/m2) was carried out on the CSC eyes.The treated eyes were examined 2 weeks, 1 month and 3 months after PDT.The BCVA,subfoveal choroid thickness,FFA and ICGA findings before and after PDT were compared.The following-up duration was 5 years or more.Results The BCVA before and 3 months after PDT were 0.5 ±0.1 and 0.9±0.2, respectively, with a statistically significant difference between them (t =19.17,P =0.00).The subfoveal choroidal thickness value 3 months after PDT was (326.56±39.47) μm,which was significantly reduced in comparison with (486.24 ±47.53) μm before PDT (t =25.17, P =0.00).FFA and ICGA showed that the leakage of fluorescein (hyperfluorescence) was disappeared in all the treated eyes.No systemic or local adverse effects and recurrence were observed during the follow-up period.Conclusions On the basis of the results of this study and available information,60％-dose verteporfin PDT seems to have a better long-term efficacy and safety than full-dose verteporfin in treating chronic CSC.

Due to the low molecular weight and simple structure, the production of specific antibodies against acrylamide is unavailable. In this study, a novel hapten was synthesized through the derivatization of acrylamide and 4-mercaptophenylacetic acid. The hapten was then coupled to carrier protein and used to immunize New Zealand rabbits. Polyclonal antibody which showed specific binding to the acrylamide derivative ( hapten) was obtained. The antibody was labeled with horseradish peroxidase ( HRP) and used to develop a direct competitive enzyme-linked immunosorbent assay ( dc-ELISA) . The dc-ELISA was used to determine the content of acrylamide derivative, and then transferred to the content of acrylamide. The assay showed an IC50 value of 45. 49 μg/L, a limit of detection of 3. 0 μg/L and the linear range of 9. 2-195 μg/L for acrylamide. The recovery of acrylamide from spiked food sample was determined ranging from 83 . 6% to 112 . 7%. Good correlations between the results of dc-ELISA and standard HPLC-MS/MS were obtained. The proposed dc-ELISA is suitable for the determination of acrylamide in food samples.

To detect furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone ( AMOZ ) in fish sample, an Eu3+ labeling time-resolved fluoroimmunoassay ( TRFIA ) was developed. The effects of experimental conditions including AMOZA-OVA concentration, dilution of antibody, and reaction time on the sensitivity of TRFIA were explored. The results showed that the optimized assay conditions were as follows:the AMOZA-OVA concentration was 0. 25 μg/mL; the antibody was diluted 5í104 folds, and the competitive reaction time was 50 min. Under optimal conditions, the method showed a detection limit of 0. 01 ng/mL, an IC50 of 0. 26 ng/mL and a linear range (IC20-IC80) of 0. 025-2. 83 ng/mL. The recoveries of AMOZ in fish at three spiked levels ranged from 78 . 0% to 86 . 0%, and the relative standard deviations were less than 15%. Good correlation between the ic-TRFIA and high performance liquid chromatography-tandem mass spectrometry was obtained for spiked food samples. The proposed ic-TRFIA method was suited for the determination of AMOZ residue in food samples.

Nowadays,the prevalence of nonalcoholic fatty liver disease(NAFLD)is constantly rising in China and globally,and its incidence rate is increasing year by year,which has seriously affected human life and health.Lipophagy is molecular chaperone-mediated autophagy and has the functions of promoting lipolysis,maintaining the lipid homeostasis of hepatocytes,and alleviating hepatocyte fatty degeneration.Lipophagy has three main processes of lipid droplet catabolism,lipid droplet autophagy,and fatty acid β-oxidation,which are regulated by key genes,receptors,and enzymes.Currently,important advances have been achieved for the intervention methods of traditional Chinese medicine,Western medicine,diet,and exercise in the research on lipophagy,which provides new perspectives for the prevention and treatment strategies for NAFLD.

Objective:To explore the effect of expected pain of blood donation puncture on actual puncture pain.Methods:A total of 350 blood donors from a blood collection site in Beijing from July 2019 to August 2019 were selected by the convenient sampling method. Blood donors evaluated the pain degree of blood donation puncture before blood test by using VAS, and VAS was used to evaluate the actual pain degree again after successful puncture. The expected pain score was compared with the perceived puncture pain score after blood donation and the influencing factors of the donors' puncture pain were analyzed.Results:The expected pain score of the 350 blood donors was (3.16±2.06) , and the perceived puncture pain score after blood donation was (2.99±1.89) . There were statistically significant differences in the expected pain assessment scores of blood donors of different genders and whether they donated blood intentionally ( P<0.05) . The interactions between gender, blood donation preparation, blood donation history, age and expected pain were statistically significant ( P<0.05) . Conclusions:Before blood donation, quality service can reduce puncture pain of female blood donors. Blood donation publicity don't need to avoid the pain of blood donation, we should improve the acceptance of puncture pain among blood donors and encourage the public to be fully prepared to participate in blood donation.

Objective To investigate the operating strategies and essentials of minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) with tubular channel (Spotlight) to treat single-level lumbar degenerative diseases. Methods From November 2013 to December 2015, 97 patients (47 males and 50 females) underwent single-level lumbar degenerative diseases fol-lowing MIS-TLIF with Spotlight were analyzed, whose age were from 35-82 years old with the average age of 57.6±12.3 years old. The preoperative diagnosis was lumbar spinal stenosis in 63 cases, lumbar spondylolisthesis in 25 cases, and lumbar instability in 9 cases. The affected level was L3,4 in 9 cases, L4,5 in 66 cases, and L5S1 in 22 cases. According to distinct clinical manifestations and radiological characteristics, different approaches of Spotlight channels were employed. Unilateral decompression via unilateral channel was performed in 52 cases, bilateral decompression via unilateral channel was performed in 22 cases, and bilateral decom-pression via bilateral channel was performed in 23 cases. Clinical outcomes included operation duration, surgical blood loss, post-operative drainage volume and complications was recorded. Average intervertebral height, lumbar and surgical Cobb angle were utilized to evaluate the reduction of intervertebral height and lumbar lordosis. The low back and leg pain were represented as Visu-al Analogue Scale (VAS) score. The preoperative and postoperative Oswestry Disability Index (ODI) score were recorded individu-ally to evaluate patients'functional recovery. Besides, the Bridwell criterion was introduced to define the extent of the lumbar fu-sion. The MacNab criterion was used for assessment of postoperative efficacy. Results The operation duration was 189.8 ± 41.3 min, the volume of surgical blood loss was 143.9 ± 102.0 ml and the volume of postoperative drainage 75.0 ± 59.0 ml in all cases. Among them, operation time was 165.0±24.2 min, surgical blood loss was 99.5±54.1 ml and postoperative drainage was 48.4±27.6 ml in the operation group of unilateral decompression via unilateral channel. The date in the group of Bilateral decompression via unilateral channel were 208.9 ± 46.0 min, 151.4 ± 96.3 ml, 88.0 ± 51.3 ml and in the group of bilateral decompression via bilateral channel were 225.4±32.0 min, 236.0±126.3 ml, 122.8±81.7 ml. All the patients were followed up for 16-42 months, the average follow-up time was 24.9 ± 7.0 months. Low back VAS reduced from 6.10 ± 0.84 preoperatively to 1.59 ± 0.49 at the final follow-up, leg VAS decreased from 6.56±0.85 preoperatively to 1.59±0.57 at the last follow-up, and ODI reduced from 59.36％±5.52％preop-eratively to 15.89％±2.90％at the final follow-up, compared with preoperative, the differences were significant. Average interverte-bral height improved from 9.92±2.25 mm preoperatively to 12.24±1.78 mm at latest follow-up time, which had statistically signifi-cant difference. Operative segment and lumbar Cobb angle were 13.81°±6.10° and 32.32°±11.97° preoperative, at the time of lat-est follow-up improved to 14.25° ± 5.57° and 35.83° ± 9.89° , Compared with preoperative, lumbar Cobb angle was significantly in-creased but operative segment Cobb had no significant difference. According to the criteria of Bridwell, intervertebral fusion at fi-nal follow-up of I and II grades were 90 cases in total (92.8％). The MacNab criteria was used to evaluate the clinical efficacy, which 69 were excellent, 23 were good, and 5 were acceptable, the excellent and good rate was 94.8％. Conclusion The tech-nique of MIS-TLIF with the tubular channel (Spotlight) is safe and efficient for the treatment of single segment lumbar degener-ative diseases. Different strategies can be selected by different preoperative clinical manifestations and radiological features.