Objective To investigate the clinical efficacy of CRRT in the treatment of severe pancreatitis.Methods 120 patients with severe pancreatitis were divided into two groups according to the treatment.All the patients were given basic treatment in all groups.The Pa arterial oxygen pressure (PaO2),oxygenation index (PaO2/FiO2),and C-reactive protein (CRP) in the two groups after 72 hours of treatment were observed.The levels of blood lactate (Lac),APACHEII,tumor necrosis factor α(TNF-α),interleukin-6 (IL-6) and interleukin-1β (IL-1β) were measured before and after treatment 72 hours.A comparison of mortality and hospitalization time was observed in 28 days of treatment.Results After treatment,Lac [the control group (3.32±0.85)mmol/L,the observation group (2.55±0.65)mmol/L],APACHEII score [the control group (13.30±2.80)points,the observation group (12.01±2.60)points],CRP [the control group (24.30±2.80)mg/L,the observation group (12.33±1.60)mg/L] were significantly lower than before treatment [Lac:the control group (4.85±1.05)mmol/L,the observation group (4.90±1.02)mmol/L;APACHEII score:the control group (16.62±2.95)points,the observation group (16.90±3.01)points;CRP:the control group (40.32±3.10)mg/L,the observation group (40.40±3.51)mg/L;the control group:tLac=1.67,P=0.004,tAPACHEII=6.32,P=0.000,tCRP=29.71,P=0.000;the observation group:tLac=15.05,P=0.005,tAPACHEII=9.52,P=0.000,tCRP=56.36,P=0.000].PaO2 [the control group (75.30±4.80)mmHg,the observation group (84.31±4.60)mmHg], PaO2/FiO2 [the control group (225.30±14.83)mmHg,the observation group (256.31±14.65)mmHg] were significantly higher than before treatment [PaO2:the control group (60.32±4.15)mmHg,the observation group (60.40±4.01)mmHg;PaO2/FiO2:the control group (130.39±11.15)mmHg,the observation group (130.90±11.01)mmHg;the control group:tPaO2=18.29,P=0.000,tPaO2/FiO2=39.62,P=0.000;the observation group:tPaO2=30.35,P=0.000,tPaO2/FiO2=53.01,P=0.000].Those in the observation group were significantly improved than the control group (tLac=5.574,P=0.00,tAPACHEII score=2.615,P=0.005,tPaO2=3.646,P=0.0002,tPaO2/FiO2=11.523,P=0.00,tCRP=28.751,P=0.000).After treatment,the TNF-α,IL-6 and IL-1β levels in the two groups [the control group:IL-1β (70.32±6.85)ng/mL,IL-6 (103.30±8.80)ng/mL,TNF-α (89.30±8.80) ng/mL;the observation group:IL-1β(48.55±6.62)ng/mL,IL-6(92.01±8.60)ng/mL,TNF-α(57.31±7.60)ng/mL] were significantly improved than before treatment [the control group:IL-1β(82.85±7.05)ng/mL,IL-6(173.62±9.95)ng/mL,TNF-α (105.32±9.15)ng/mL;the observation group:IL-1β(83.90±7.32)ng/mL,IL-6 (175.90±10.01)ng/mL,TNF-α (106.40±9.01)ng/mL;the control group:tIL-1β=9.66,P=0.000,tIL-6=41.01,P=0.000,tTNF-α=9.77,P=0.000;the observation group:tIL-1β=27.74,P=0.000,tIL-6=49.23,P=0.000,tTNF-α=32.26,P=0.000].And those of the observation group improved more significantly than the control group (tIL-1β=17.702,P=0.00,tIL-6=7.107,P=0.00,tTNF-α=21.311,P=0.000).The mortality rate and hospitalization time of the observation group were 18.33% and (10.97±2.92)days,which were significantly lower than those of the control group [36.67%,(13.63±3.26)days;χ2=5.058,P=0.025;t=4.708,P=0.000).Conclusion The use of CRRT in the treatment of severe pancreatitis can improve the vital signs,reduce the inflammation index,improve the serum levels of inflammatory factors and lactic acid,reduce the mortality and hospital stay.

BACKGROUND: Bone marrow mesenchymal stem cells(MSCs) have been investigated initially. Because both fat tissue and bone marrow are tissues originated from mesoderm, so whether MSCs could be obtained from fat tissue as well, which also have multi-lineage differentiation potential?OBJECTIVE:To investigate the basic biological characteristics of adipose MSC and its differentiation into osteoblast under given culture condition for the exploration of its feasibility as seed cell in bone tissue engineering.DESIGN: A randomized and controlled study based on cells.SETTING: Department of Orthopaedics of an Affiliated Hospital of a university.MATERIALS: The study was conducted in the Department of Orthopaedics of the Third Hospital of Peking University. Adipose MSC extracted from the fat tissue of Lewis rats was used as subject.METHODS: Adipose MSCs were obtained from inguinal fat pads of Lewis rat after digestion, which were induced into adipocytes and osteoblasts with adipose and osteogenesis induced culture mediums. The differentiations were examined with cytochemical staining, immuncytochemical staining and Western blotting.MAIN OUTCOME MEASURES: Morphological and biological characteristic of adipose MSCs, and the specific mark of osteoblast after induction.RESULTS: Adipose MSCs were obtained from rat adipose tissue culture,which appeared fibroblast-likely in the culture in vitro, and could stably proliferate and passage in vitro. Primary adipose MSCs could differentiate into adipocytes spontaneously, and passaged cells could form fat drop under the reaction of insulin and dexamethasone, and then differentiate towards adipocytes after the expression of peroxidase proliferation activated receptor ?(PPAR-?) enhanced. There was significant difference between induction group and control group in alkaline phosphatase(ALP) activity detection under the induction of dexamethasone, ascorbic acid, and ?-sodium glycerophosphate(P ＜ 0.01) . Calcium node appeared in yon Kossa staining. Result of osteopontin (OPN) immunocytochemcial staining was positive,and OPN expression was detected by Western blotting after induction.CONCLUSION: MSCs with multi-lineage potential can be obtained from rat adipose tissue, and differentiate into adipocytes and osteoblasts after inductions. Therefore, adipose MSCs can possibly be served as one of optimal seed cells for bone tissue engineering.

BACKGROUND: Oxygen-glucose depdvation (OGD) in cultured neurons simulates stroke to a certain degree and plays an important role in studying processing and pathophysiological mechanism of ischemic neuronal injury.OBJECTIVE: To produce experimental OGD models in cultured neurons.DESIGN, TIME AND SETTING: A grouping controlled study was performed at the Center Laboratory of Third Hospital, Peking University from January 2007 to March 2008.MATERIALS: Fetal Wistar rats with gestational age of 17-19 days were collected in this study.METHODS: Primary cultures of cortical neurons that were derived from fetal Wistar rats with gestational age of 17-19 days were performed to remove pollutional non-neuronal cells. OGD was produced by incubation with non-glucose balanced salt solution and 2% Oxyrasa in 7-day cultured cortical neuron cultures. Cell cultures were kept in a humidified 37 ℃ incubator. In the control group, cell culture medium was replaced with balanced salt solution containing 20 mmol/L glucose. In the sham operation group,balanced salt solution containing 20 mmol/L glucose and Oxyrasawere used to replace the medium.MAIN OUTCOME MEASURES: Oxygen concentration in the culture medium was measured with blood gas analysis; neuronal death in the experimental group was observed under phase contrast microscope; lactate dehydrogenase activity was detected with lactate dehydrogenasa assay; effect of oxygen-glucose deprivation on neuronal viability was observed with trypan blue staining.RESULTS: Measurement of oxygen concentration showed that hypoxia could be quickly achieved shortly after the addition of Oxyrase; lactate dehydrogenase assay revealed that after treatments of neuron cultures with Oxyrase and non-glucose balanced salt solution, lactate dehydrogenase release increased significantly with the treatment time; trypan blue staining and phase contrast microscope showed that cell viability decreased after treatments of Oxyrase and non-glucose balanced salt solution, and most neurons died 6 hours after OGD.CONCLUSION: These results show that Oxyrase, together with non-glucose balanced salt solution, can be conveniently used to produce OGD condition in cultured neuronal cells which is greatly useful in the study of simulating cerebral ischemia in vitro.

Aim To investigate the neuroprotective effect of verbascoside,one of the phenylethanoids isolated from the stems of Cistanche salsa,on MPP+-induced injury in SHSY5Y cells.Methods SHSY5Y cells were exposed to various doses of verbascoside for 12 h,and then treated with 200 ?mol?L-1 MPP+ for 24 h.The cell viability was observed with MTT assay;reactive oxygen species,the mitochondrial membrane potential and the percentage of apoptosis were measured by flowcytometer;the activation of caspase-3 was measured with the caspase-3 activity assay kit;the expression of Bcl-2 was measured with Western blot.Results Following treatment with MPP+ for 24 h,MPP+ induced a significant decrease of cell viability;apoptosis percentage were 38.9%;accumulation of intracellular ROS,increase of caspase-3 activity and the decreases in mitochondrial membrane potential were detected.However,pretreatment with verbascoside (0.1,1 or 10 mg?L-1) for 12 h exhibited cytoprotective effects in a dose-dependent manner.Verbascoside obviously enhanced cell viability,and significantly reduced the number of cells labeled with Annexin-V.The percentage of apoptosis neurons was significantly decreased to 29.5%,15.3% and 8.6% respectively.Flowcytometer showed the verbascoside attenuated the accumulation of ROS and the MPP+-induced collapse of mitochondrial membrane potential in SHSY5Y cells.And significant decreases were detected in caspase-3 activity compared with the MPP+-treated cells at the same time point.Moreover,pretreatment with verbascoside promoted the expression of Bcl-2.Conclusions verbascoside had the neuroprotective capacity to antagonize MPP+-induced apoptosis in SHSY5Y cells,and might be useful in treating Parkinson disease.

AIM: To investigate the neuroprotective effect of tubuloside B,one of the phenylethanoids isolated from the stems of Cistanche salsa,on H2O2-induced injury in PC12 cells.METHODS: PC12 cells were exposed to various doses of tubuloside B for 12 h,then treated with H2O2 at concentration of 100 ?mol/L for 24 h.The cell viability was observed with MTT assay.Reactive oxygen species and the mitochondrial membrane potential were measured with laser scanning confocal microscopy(LSCM).The DNA content and percentage of apoptosis were assayed by DNA agarose gel electrophoresis and flow cytometry.The activation of caspase-3 was detected with the caspase-3 activity assay kit.RESULTS: Following treatment with H2O2 for 24 h,H2O2 induced a significant decrease in cell viability;DNA ladder was observed and apoptosis percentage was as high as 48.0%.Accumulation of intracellular ROS,increase in caspase-3 activity and the decrease in mitochondrial membrane potential as indicated with the decrease of red/green ratios(from 5.97 to 0.41) were detected.However,pretreatment with tubuloside B(1,10 or 100 mg?L-1) for 12 h exhibited cytoprotective effects in a dose-dependent manner.Tubuloside B obviously enhanced the cell viability,reduced formation of the DNA ladder,and significantly reduced the number of cells labeled with Annexin-V.The percentage of apoptosis/necrosis neurons was significantly decreased to 30.9%,18.3% and 6.2%,respectively.LSCM showed that the tubuloside B attenuated the accumulation of ROS and the H2O2-induced collapse of mitochondrial membrane potential in PC12 cells.The significant decrease in caspase-3 activity was detected,compared to the H2O2-treated cells at the same time point.CONCLUSION: Tubuloside B has the neuroprotective capacity to antagonize H2O2-induced apoptosis and injury in PC12 cells,indicating it may be useful for treating some neurodegenerative diseases.

Objective The aim is to observe the role and mechanism of estradiol ( E2 ) on the proliferation of rat hippocampal neural stem cells ( NSCs ) .Methods Twenty hippocampi from embryonic 17-day ( E17 ) SD rats were dissociated and plated into culture flasks with NSCs specific medium containing different concentrations of estradiol .The proliferation and the vitality of NSCs were detected by immunofluorescence against BrdU and MTT assay .The expression of estrogen receptors ( ERαand ERβ) was measured by immunofluorescence staining combined with Nestin double labeling . Results BrdU and MTT assay results showed that the cell number increased when the concentration of estradiol increased from 10 -10 to 10 -8 mol/L.The number of cell proliferation and the viability of cells were best at the concentration of 10 -8 mol/L compared to the other groups .However, when the estradiol concentration was increased from 10-8 to 10 -6 mol/L, the cell proliferative capacity declined gradually .Double immunofluorescence labeling showed that the two types of estrogen receptors ( ERαand ERβ) were expressed in the cultured hippocampal NSCs .Conclusion Estradiol promotes the proliferation of hippocampal NSCs in a certain concentration range , and ERαand ERβmay be involved in the estradiol-induced proliferation .

Objective To evaluate the effects of fluid restriction in combination with small dose of norepinephrine on cerebral oxygen metabolism in elderly patients undergoing gastrointestinal surgery.Methods Forty elderly patients of both sexes,aged 65-80 yr,with body mass index of 18-24 kg/m2,of ASA physical status Ⅰ or Ⅱ (NYHA Ⅰ or Ⅱ),with left ventricular ejection fraction≥50％,undergoing elective gastrointestinal surgery,were randomly divided into 2 groups (n =20 each) using a random number table:routine fluid administration group (group S) and restricted fluid administration + small dose of norepinephrine group (group RN).In group S,lactated Ringer's solution was given routinely,ephedrine 5 mg (per time) was injected intravenously,and MAP was maintained ≥ 65 mmHg during operation.In group RN,lactated Ringer's solution was infused intravenously at 5 ml · kg-1 · h-1 starting from 30 min before anesthesia,norepinephrine was infused intravenously at 0.01-0.03 μg · kg-1 · min-1 after induction of anesthesia,and MAP was maintained ≥ 65 mmHg.Intraoperative blood loss was replaced with the equal volume of 6％ hydroxyethyl starch 130/0.4 sodium chloride injection in both groups.At 5 min before skin incision,1 and 2 h after skin incision and postanesthesia care unit discharge time,arterial and jugular bulb venous blood samples were obtained for blood gas analysis,and arterial oxygen content,jugular bulb venous oxygen content,arteriovenous oxygen content difference,cerebral oxygen extraction rate,and the ratio of cerebral blood flow to cerebral oxygen metabolic rate were calculated.Results There were no significant differences between the two groups in arterial oxygen content,jugular bulb venous oxygen content,arteriovenous oxygen content difference,cerebral oxygen extraction rate,and the ratio of cerebral blood flow to cerebral oxygen metabolic rate.Conclusion Fluid restriction combined with small dose of norepinephrine produces no effects on cerebral oxygen metabolism in elderly patients undergoing gastrointestinal surgery.

Objective:To investigate the roles of tacrolimus pretreatment on hepatic ischemia reperfusion injury (IRI) and its possible mechanism in a rat autologous orthotopic liver transplantation (AOLT) model.Methods:For 24 specific pathogen free 8-10 week male Sprague Dawley rats (220-250g) were randomly and equally divided into three groups. The abdomen of sham-operated group was only opened and closed; the treatment with tacrolimus was administered via dorsal penile vein before the experiment in tacrolimus-pretreated group; the AOLT group and tacrolimus-pretreated group were set to construct the AOLT IRI rat models. The levels of ALT, AST, tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β) in serum were tested after the reperfusion. The change of liver structure was evaluated by H&E staining. The quantitative real-time PCR (RT-qPCR) and Western blot assay were used to test the mRNA and protein level of hypoxia-inducible factor-1α (HIF-1α) and heme oxygenase-1 (HO-1).Results:The levels of serum ALT (1 332.0±52.8) U/L and AST (2 472.0±257.8) U/L in the AOLT group were higher than the levels in the sham-operated group (65.0±17.4)U/L, (222.3±45.2) U/L and tacrolimus-pretreated group (789.9±54.0) U/L, (533.4±31.6) U/L. The differences were significant ( P<0.05). And in the tacrolimus-pretreated group there were less lesions in the liver than in the AOLT group. The serum level of TNF-α and IL-1β of the AOLT group were increased than the sham-operated group and tacrolimus-pretreated group, and the differences were significant ( P<0.05). Compared with the AOLT group, the expressions of HIF-1α and HO-1 were increased significantly after the tacrolimus pretreatment ( P<0.05). Conclusion:Tacrolimus pretreatment could reduce rats hepatic cold IRI by inducing the expressions of HIF-1α and HO-1, and inhibiting the production of inflammatory cytokines.

Objective To evaluate the clinical effect of arthroscopic treatment of femoroacetabular impingement syndrome(FAI) for patients over 50 years old,and explore the occurrence regularity and treating rules for such a disease.Methods The clinical data of 71 patients(78 hips with FAI) over 50 years old who underwent arthroscopic treatment for FAI in our department between May 2012 and May 2017 were studied retrospectively.Physical examination,X-ray and CT 3D scans were made preoperatively for explicit diagnosis.The follow-up period ranged from 6 to 66 months,with an average of 31.78 ± 18.07 months.Every patient had a joint space greater than 2 mm,and a grade Ⅰ or Ⅱ hip osteoarthrosis according to the Trnnis scale.Under the hip arthroscopy the synovial hyperplasia was cleaned,the damaged labrum and cartilage were repaired,and the femoroacetabular hyperplasia and the impingement factors were removed to restore the normal shape of femoroacetabular.The joint clearance,changes ofthe alpha angle when at Dunn position and centre edge(CE) angle at normotopia on the X-ray,the intraoperative injuries of cartilages and glenoid labrum and surgical satisfaction and complications were measured and recorded.The modified Harris hip score(mHHS) and visual analogue scale(VAS) were used to evaluate the hip function recovery and pain relief of patients.Results The average age of the patients was 55.15 ± 5.02 years old,ranging from 50 to 69.Among the 23 males and 48 females,there were 33 left hips with FAI and 45 right hips with FAI.The average preoperative joint clearance was 4.81 ± 0.87 mm,and all incisions were healed by first intention after the treatment.The average α angle of the patients decreased from 50.11 ± 4.75 to 42.72 ± 4.7 degrees after the treatment,with the α angle of 7 patients(8.97％) bigger than 55 degree,and that of 40 patients(51.28％) smaller than 50 degree.The average CE angle decreased from 36.54 ± 9.14 degrees to 35.19 ± 8.55 degrees after the treatment,with that of 27 patients(34.62％) bigger than 40 degrees.Before the treatment,the main clinical manifestations were hip pain and swelling,including 36 cases(46.15％) with hip joint lock,70 (89.74％) with groin tenderness.Moreover,75 cases(96.15％) were positive in hip adduction internal rotation test(FADIR) and 64 cases(83.33％) were positive in the hip abduction and external rotation impingement test.However,the pain was relieved or disappeared after the treatment.The average VAS score decreased significantly from preoperative 4.42 ± 1.42 points to 1.31 ± 1.28 at the last follow-up,while the average mHHS score increased significantly from preoperative(52.4 ± 19.38) points to(81.72 ± 10.82) during the last follow-up,and the difference was statistically significant (P＜0.01).Significant improvement was observed in the mHHs and VAS scores of 89％ patients(P＜0.01).1 patient(1.28％) underwent hip replacement during the follow-up period.No serious complications occurred.Conclusion In most cases,arthroscopic treatment of FAI in old patients(over 50 years old),who were with osteoarthrosis and hip labrum injury,can significantly improve the joint function and relieve pain.It is a treatment with safety.

Objective: To investigate the effect of DAMP (damaged associated molecular pattern) on the inhibition of RAS-mutant A549 lung adenocarcinoma cells by CIK cells and its mechanism. Methods: Human peripheral blood mononuclear cells were isolated in vitro and CIK cells were cultured. A549 cells were treated with cisplatin (DDP) and doxorubicin (ADM) alone or in combination, and the morphology of A549 cells was observed under a microscope. The supernatant of A549 cells was co-cultured with CIK cells. Flow cytometry was used to detect the CIK cell immunophenotype after co-culture. MTT assay was used to detect the inhibition of A549 lung cancer cell proliferation induced by A549 cell supernatant. The concentration of chemotherapeutic drugs kills A549 cell supernatant CRT, ATP, HMGB1 content. Results: Low-level chemotherapeutic drugs showed more immunogenic death characteristics after killingA549 cells. The ratio of CD8+ and CD56+ in CIK cells was significantly higher than that in control CIK cells (P<0.05). The inhibition rate of CIK cells induced byA549 cells after injury onA549 lung adenocarcinoma cells was significantly higher than that of the same dose chemotherapy group [DDP group (31.34±1.51)% vs (5.97±1.74)%, ADM group (45.46±1.78)% vs (6.22±1.34)%, DDP+ ADM group (45.78±1.14)% vs (11.94±3.11)%, all P<0.05], and low-mass chemotherapeutic agents killed C549 induced by A549 cell supernatant on A549 The inhibition rate of the cells was higherthan that of the supernatant induced by the higher concentration of chemotherapeutic drugs (all P<0.05). The level of CRT,ATP, and HMGB1 in immunogenicity-related molecules in the supernatant ofA549 cells was significantly increased by low-concentration chemotherapy drugs (all P<0.05). In the low-concentration group, the supernatant-induced inhibition of the proliferation of A549 lung adenocarcinoma cells increased with the increase of CRT, ATP, and HMGB1 levels. Conclusion: The combination of lower concentration of DDP and ADM alone or in combination could more easily induce the immunogenic death of A549 cells and release higher levels of DAMP molecules, which could promote the inhibitory effect of CIK on lung cancerA549 cells.