OBJECTIVE To evaluate the possibility that CdCl2 induces autophagy and apoptosis in HEK293 cells,and the role of extracellular regulated protein kinases (ERK1/2) and AKT proteins in autophagy. METHODS Green fluorescence protein(GFP)-light chain 3B(LC3B)expression plasmid was transfected into HEK293 cells. After 24 h,HEK293 cells were induced with CdCl2 2,4,8 and 10μmol·L-1 for 12 h. The expression of GFP-LC3B was detected by fluorescent microscopy. HEK293 cells were induced with CdCl2 2,4,8 and 10μmol · L-1 without transfection of GFP-LC3B for 12 h while autophagic vacuoles were observed by transmission electron microscopy. The expression of LC3B-Ⅱ/Ⅰproteins and the phosphorylation levels of ERK1/2 and AKT were analyzed by Western blotting. Apoptosis was detected by flow cytometry microscopy. HEK293 cells were treated with 3-MA 20μmol · L-1+CdCl2 10 μmol · L-1 for 12 h before cleaved caspase 3 protein was detected by Western blotting. RESULTS When HEK293 cells were exposed to CdCl2(≤10μmol · L-1)for 12 h,cytoplasmic GFP-LC3B punctuates were observed under the fluorescence microscope,and autophagic vacuoles were observed under an electron microscope. The expression of LC3B-Ⅱ/Ⅰ,p-ERK1/2 and p-AKT proteins was significantly increased in CdCl2-induced cells(P<0.05,P<0.01). Moreover,apoptosis was observed. The addition of 3-MA 20μmol · L-1+CdCl2 10μmol · L-1 enhanced apoptosis. Cleaved capase 3 protein expression was significantly increased(P<0.01). CONCLUSION CdCl2(≤10μmol·L-1)can induce autophagy in HEK293 cells. ERK1/2 and AKT proteins might be associated with the activation of autophagy that is accompanied by apoptosis,suggesting that autophagy can inhibit apoptosis at certain concentrations of CdCl2.

OBJECTIVE:To compare clinical efficacy and safety of intravenous dripping of methotrexate and uterine arterial perfusion embolization combined with complete curettage of uterine cavity (CCUC) in the treatment of cesarean scar pregnancy (CSP).METHODS:A total of 90 CSP patients were randomly divided into group A and B,with 45 cases in each group.Group A was given Methotrexate (MTX) injection 50 mg/m2 intravenously before CCUC.Group B received Seldinger catheterization in supine position before CCUC,and was given sequential infusion of MTX injection 50 mg/m2 and gelatin sponge particles into the uterine artery;the catheter was removed after satisfactory embolization by imaging examination.The levels of β-HCG of 2 groups were reexamined every 24 h,and CCUC was performed when serum β-HCG level was below 1 000 mU/mL.Perioperative bleeding volume,postoperative bleeding volume,the time of blood β-HCG returning to normal,the time of menstruation returning to normal,hospitalization time as well as the lesion diameters,the levels of blood β-HCG,the occurrence of compliance and ADR before and after treatment were compared between 2 groups.RESULTS:The intraoperative bleeding amount and postoperative bleeding amount of group B were significantly lower than those of group A,while the time of blood β-HCG returning to normal,the time of menstruation returning to normal and hospitalization time were significantly shorter than group A,and the incidence of total compliance was significantly lower than group A,with statistical significance (P＜0.05).Before medication,there was no statistical significance in lesion diameters between 2 groups (P＞0.05).Before CCUA,the lesion diameters of 2 groups were significantly smaller than before medication,and the group B was significantly smaller than group A,with statistical significance (P＜0.05).Before medication,there was no statistical significance in blood β-HCG levels between 2 groups (P＞0.05).Before and after CCUC,blood ββHCG levels of 2 groups were significantly lower than before medication,and the group B was significantly lower than group A,with statistical significance (P＜0.05).There was no statistical significance in the incidence of ADR between 2 groups (P＞0.05).CONCLUSIONS:Compared with methotrexate by intravenous drip,methotrexat by uterine artery embolization combined with CCUC for CSP can effectively reduce intraoperative bleeding amount,promote the decrease of blood β-HCG and menstrual recovery,and contribute to the reduction of complication risk with good safety.

Background Penetrating keratoplasty (PKP) has become an effective method of treatment for fungal keratitis in recent years,but the application timing of glucocorticoids after PKP is still unclear.Literature reported that the concentration of tear (1,3)-β-D-glucan in fungal keratitis was significantly higher than that in normal.Objective This study was to investigate the change of tear (1,3)-β-D-glucan before and after PKP in fungal keratitis and to explore the application duration of anti-fungal drugs and application timing of glucocorticoids.Methods This study protocol was approved by ethic committee of Affiliated Hospital of Qingdao University.A serial cases-observational study was performed from August,2011 to December,2012.Twenty eyes of 20 patients with fungal keratitis were collected in Affiliated Hospital of Qingdao University.PKP was performed in affected eyes,and the fellow health eyes served as controls.Tear of 50 μl was obtained in the controls on 1 day before operation and 1 day,7,14,21 and 28 days after operation to detect tear (1,3)-β-D-glucan levels.Results Tear (1,3)-β-D-glucan levels were (14.67±3.84)mg/L,(1 861.66±196.17) mg/L,(927.71±155.82)mg/L,(392.30±71.22)mg/L,(179.60±40.47) mg/L,(40.20± 12.46) mg/L and (15.12± 1.80) mg/L in the control group,preoperative 1 day,postoperative 1 day,7,14,21,28 days,respectively,showing a significant difference among various time points (F=883.45,P=0.00).Tear (1,3)-β3-D-glucan levels were gradually reduced with the lapse of the postoperative time,with significant differences between adjacent timepoints (t' =13.84,t =16.67,t' =11.02,t' =13.97,t' =-8.45,all at P=0.00).Tear (1,3)-β-D-glucan levels in postoperative 28 days came near that of normal control group,without significant difference between them (P =0.64).Fungal keratitis recurred in 2 eyes on the fifth and sixth day after operation,with the tear (1,3)-β-D-glucan levels of 2 350.24 mg/L and 1 992.82 mg/L,respectively.Conclusions The concentration of (1,3)-β-D-glucan in the tears increases in the eyes with fungal keratitis and drops to normal range at 28 days after PKP,indicating that the antifungal eyedrops should be applied until 4 weeks after PKP,and this is an optimal timing of using corticosteroid eyedrops to resist reject reaction.

The accurate location of acupoints is the prerequisite for the efficacy of acupuncture-moxibustion. Bone-length measurement, bone-proportional measurement, and proportional unit of the body measurement are commonly used in clinic for acupoints location. Based on the systematic review of the ancient literatures, this article discussed the meaning, evolution and relations of bone-length measurement, bone-proportional measurement, and proportional unit of the body measurement. It’s concluded that the bone-length measurement should be a benchmark in ancient anthropometry and an important basis for acupoints location. Bone proportional measurement, used in different genders, ages and body sizes, was based on the relatively stable proportional relations of various body parts, though some of the standards were adjusted in accordance with the correlation between meridians and acupoints. Location of points by proportional unit of the body simplified the application of bone-length measurement and bone proportional measurement, based on the ratio between short and long bones or the ratio of same body parts in anthropometry. However, proportional unit of the body measurement should be applied for the specific body parts. Bone-length measurement, bone-proportional measurement, and proportional unit of the body measurement are correspondingly the benchmark measurement, relative measurement, and simplified measurement.

Inflammation plays an important role in the pathophysiological mechanism of acute ischemic stroke.CD200 expressed in neurons interacts with CD200 receptor (CD200R) on microglia cells.It can inhibit microglia activation and alleviate the inflammation after cerebral ischemic injury.This article reviews the roles of CD200 and CD200R in the activation of microglia after cerebral ischemia.

Objective To compare the cerebral protective effect of propofol and sevoflurane combined with sufentanil anesthesia in the patients undergoing valvular surgery under cardiopulmonary bypass (CPB).Methods Sixty American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients, aged 60-70 yr, scheduled for elective valvular surgery under CPB, were randomly divided into either propofol-based anesthesia group (group PA) or sevoflurane-based anesthesia group (group SA) , with 30 patients in each group.From induction of anesthesia to the end of surgery, group P received targetcontrolled infusion of propofol 0.5-2.0 μg/ml, and group S continuously inhaled 0.5％-2.5％ sevoflurane.Bispectral index value was maintained at 45-55.Immediately after induction (T0), at the end of surgery (T1) , and at 6, 12 and 24 h after surgery (T2-4) , the superior vena cava was retrogradely cannulated for blood sampling, and the concentrations of plasma S-100β protein and neuron-specific enzyme were determined using enzyme-linked immunosorbent assay.Results Compared with group SA, the plasma S-100β concentrations at T1,2 and neuron-specific enzyme concentrations at T1-3 were significantly decreased in group PA.Conclusion The cerebral protective effect of propofol combined with sufentanil anesthesia is superior to that of sevoflurane combined with sufentanil anesthesia in the patients undergoing valvular surgery under CPB.

Objective To observe the effects of different anesthetic solutions on postoperative cognitive function and serum S100β protein levels.Methods A prospective randomized controlled trial was conducted. Ninety patients necessary to perform off-pump coronary artery bypass grafting (op-CABG) in Tianjin Chest Hospital from November 2013 to July 2014 were enrolled. They were divided into three groups by random number table: P1, P2 and P3 groups, 30 cases in each group. The anesthesia was maintained with propofol by target-controlled infusion (TCI) in all the patients in the three groups, and the respective dosages were < 2.0μg/mL, 2.0 - 3.0μg/mL and > 3.0 - 4.0μg/mL. The operation time, anesthesia time, dosages of propofol and the incidence of postoperative cognitive dysfunction (POCD) were compared among the three groups. At the following times: before induction of anesthesia (T0), completion of anesthesia induction (T1), after tracheal intubation (T2), skin incision (T3), 1 hour after operation (T4), 2 hours after operation (T5) and the end of operation (T6), the narcotrend index (NTI) and hemodynamic levels were observed; the serum S100β protein levels were measured at the following times: before induction of anesthesia (Ta), 2 hours after operation (Tb), the end of operation (Tc), postoperative 6 hours (Td) and postoperative 24 hours (Te).Results There were no significant differences in operation times and anesthesia times among three groups (allP > 0.05); dosages of propofol in P2 and P3 groups were obviously higher than those of P1 group (mg: 1 746.3±43.9, 2 332.7±42.8 vs. 968.5±35.6, bothP < 0.05), and the incidences of POCD in P2 and P3 groups were lower than that in P1 group (10.00%, 6.67% vs. 33.33%, bothP < 0.05). With the extension of anesthesia time, the level of NTI was gradually declined in each group, in P3 group, it was occasionally increased at T6, and beginning from time point T1 afterwards, the NTI levels were lower than those of P1 and P2 groups at all the time points (allP < 0.05); the mean arterial pressure (MAP) in the three groups had a tendency of firstly going down and then increasing, and the inflection point being at T2, in P1 group, the elevation of MAP level persisted to T4, and it began to decline at T5, while in P2 and P3 groups, the levels started to decline at T4; the heart rate (HR) in three groups showed a tendency of firstly going up and then declining, and the inflection point being at T3; until T6, in P3 group, MAP and HR were all lower than those of P1 and P2 groups, the differences being statistically significant [MAP (mmHg, 1 mmHg = 0.133 kPa): 74.9±8.3 vs. 85.3±11.2, 84.2±10.1;HR (bpm): 74.1±4.2 vs. 80.9±8.1, 78.7±7.9, allP < 0.05]. The serum S100β protein levels of three groups at Tb began to be obviously higher than those at Ta, and reached the peak points at Tc, then the levels started to decline until Td, and the levels at Te was approximately close to those of Ta, but the serum S100β protein levels in P2 and P3 groups were lower than that in P1 group, the differences being statistically significant (mg/L: 1.05±0.22, 1.04±0.21 vs. 1.33±0.22, bothP < 0.05).Conclusion Application of propofol by TCI 2.0 - 3.0μg/mL for maintenance of anesthesia can achieve the satisfactory depth of anesthesia, and it not only can reduce the effects on hemodynamics, but also can decrease the serum S100β protein level and the incidence of POCD.

Objective To investigate the effects of grape seed proanthocyanidin(GSPE) on mitochondrial injury in hippocampus and learning-memory impairment after obstructive sleep apnea hypoxia in rats.Methods Male SD rats(n=80) were randomly divided into control group,model group,low dose of GSPE treatment group and high dose of GSPE treatment group.Rats in control group were exposed in air,the model group were suffered from intermittent hypoxia conditions (50 ml/L,8-hour-intermittent hypoxia everyday,and the duration of experiment 2 and 6 weeks,respectively).Mitochondrion pathology in hippocampal region was observed using electron microscope;malondialdehyde (MDA) contents and superoxide dismutase activity were detected by colorimetry and apoptotic cells was measured by TUNEL method.The cognitive function of rats in each group was assessed with the Morris water maze (MWM).Results After hypoxia,mitochondrion was significantly injured.The MDA contents were increased(79.86 ± 2.52,88.26 ± 2.86) and SOD level decreased (70.67 ± 6.70,64.26 ± 7.86).The number of neural apoptotic cells was significantly enhanced (9.68 ± 0.79,15.9 ± 2.92).MWM test showed that the escaping latency was prolonged and the frequency of crossing the platform was decreased (P ＜ 0.05).Compared with that in the model group,low dose of GSPE decreased MDA contents (76.38 ± 1.96,82.16 ±2.02),increased SOD level(76.20 ± 6.86,70.58 ± 6.86),and decreased apoptotic cells (6.60 ± 0.69,9.54 ±1.36).MWM test showed that the escaping latency was shortened and the frequency of crossing the platform was increased in GSPE treatment groups(P ＜ 0.05).Compared with low dose of GSPE,high dose of GSPE decreased MDA contents increased SOD level and decreased apoptotic cells.MWM test showed that the escaping latency was shortened and the frequency of crossing the platform was increased (P＜ 0.05).Conclusion GSPE can attenuate mitochondrial injury and improve learning-memory function after obstructive sleep apnea hypoxia.

Objective To explore the clinical significance of a series of cytokines and peripheral blood immunocyte subsets before and after immunosuppressive therapy in patients with immune thrombocytopenia (ITP).Methods The percentages of immunocyte subsets in the peripheral blood of 20 patients with ITP and 20 healthy controls were detected by flow cytometry,including CD3+,CD4+,CD8+,CD4+/CD8+,CD1~.ELISA was applied to detect the level of serum TNFα,IL-2,IL-6,IL-4,IL-10,IL-11,IL-17,IL-27,transforming growth factor β (TGFβ),thrombopoietin (TPO) of 20 patients with ITP and 20 healthy controls.Results The percentage of CD3+ T lymphocyte,CD4+ T lymphocyte and the ratio of CD4+ / CD8+ T lymphocyte in patients with ITP were lower than those in healthy controls [(62.66 ± 6.58) ％ vs (69.93 ± 4.81) ％,(29.46 ± 5.02) ％ vs (39.08 ± 3.50) ％,0.97 ± 0.35 vs 1.56 ± 0.26,all P ＜ 0.05].After immunosuppressive therapy,the percentage of CD3+ T lymphocyte,CD4+ T lymphocyte and the ratio of CD4+/CD8+ T lymphocyte [(71.49 ±5.16)％,(39.25 ±3.21)％ and 1.56 ±0.28] recovered to the same levels in healthy controls.The percentage of CD8+ T lymphocyte and CD19+ B lymphocyte in patients with ITP were higher than those in the healthy controls [(30.28 ±4.63)％ vs (25.90±3.06)％,(18.92 ± 4.27)％ vs (13.17 ± 3.64)％,all P ＜ 0.05].After treatment of immunosuppressive therapy,the percentage of CD8+ T lymphocyte and CD19+ B lymphocyte [(25.16 ± 3.45) ％ and (11.98 ± 3.68) ％] recovered to the similar levels in healthy controls.The serum levels of IL-4,IL-6,IL-11,IL-17 and TPO in patients with ITP were significantly higher than those in healthy controls.While TGFβ level was significantly decreased.There was no significant difference of IL-27 between ITP patients and healthy controls.After the treatment of immunosuppressive therapy,IL-4,IL-6,IL-11,IL-17,TPO and TGFβ were down-regulated while IL-27 was up-regulated.There was no significant difference of IFNγ,TNFα,IL-2 and IL-10 among ITP patients before or after immunosuppressive therapy and healthy controls.Conclusions The present study suggests that the aberrant immunocyte subsets and cytokines are involved in the pathogenesis of ITP.Hyper-function of Th2 and Th17,dysfunction of Treg cells,up-regulation of IL-27,IL-11,TPO and other factors may contribute to the pathogenesis of ITP.

Objective To evaluate the clinical effect of Tanreqing in the treatment of radiation pneumonia. Methods 75 patients with radiation pneumonia were randomly divided into treatment group and control group according to the registration of this study.38 cases in the treatment group treated by Tanreqing,antibiotics and glucocorticoid,while 37 cases in the control group was given antibiotics and glucocorticoid.Then,the therapeutic efficacy was compared.Results From the chest routine scan result and the clinical symptom release,the excellence rate of the treatment group was 68.4%,which of the control group was 43.2%,there was statistically significant difference between the two groups (χ2 =4.823,P =0.028 ).Conclusion Treating radiation pneumonia with Tanreqing has good effect,and it is worthy of clinical application.