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This study explores the effects of enhancing the definitive hematopoiesis(DH)signals during the differentiation of human embryonic stem cells(hESCs)into hematopoietic stem/progenitor cells on the generation efficiency and effector function of natural killer(NK)cells generated from hESCs(also known as hESC-NK cells).The hESC(H1)were transformed into embryoid bodies(Ebs)by centrifugation,and during the induction of K562,were used to analyze the efficiency of hematopoietic differentiation,the efficiency of NK cell generation from hESC,the in vitro effector functions,and the expression of effector function related surface receptors.Compared to the control group,the DH group had a significant increase in the number of arterial hematopoietic endothelial cells(CD34+DLL4+)and a significant decrease in primitive hematopoietic related cells(CD34-CD43+)on day 8 of hematopoietic differentiation(P<0.05).On day 28 of NK cell differentiation,the DH group demonstrated a significant increase in the number of NK cells(CD45+CD56+),while a slight increase in the expression of effector function-related molecules such as IFN-γ,Granzyme B,Perforin and CD107a without statistical significance.Furthermore,the activation receptors CD16a and CD69 were significantly increased,NKP46 was significantly decreased,the inhibitory receptor NKG2A was significantly increased,while CD96 was significantly decreased on hESC-NK cells of DH groups(P<0.05).Conclusively,enhancing the signals for definitive hematopoiesis during hESC differentiation into hematopoietic stem/progenitor cells significantly improves the yield of NK cells and the expression of CD16a without affecting their in vitro effector functions.Our study provides a new approach to improving the efficiency of hESC-NK cell or iPSC-NK cell generation.
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Objective To explore the prognostic values of telomerase reverse transcriptase promoter (TERTp) mutation and 1p/19q co-deletion in newly-diagnosed O6-methylguanine-DNA methyltransferase (MGMT) promoter un-methylated/isocitrate dehydrogenase (IDH) wild-type glioblastoma multiform (GBM). Methods A total of 82 patients pathologically newly-diagnosed MGMT promoter un-methylated/IDH wild-type GBM, admitted to our hospitals from March 2016 to November 2018, were included in this study. TERTp mutations (TERTp wild-type and TERTp mutation [C228 mutation and C250 mutation]) in GBM specimens were detected by PCR sequencing, 1p/19q co-deletion in GBM specimens was detected by fluorescence in situ hybridization (FISH), and clinical data, adverse reactions and prognoses of patients with different molecular typing were compared. Results There were 33 patients in the TERTp wild type group with mean age of 48 years, and 49 patients in the TERTp mutation group with mean age of 59 years; the difference of age was significant (P<0.05); there were no statistical differences in gender distribution, Karnofsky performance status (KPS) scores, tumor sites and surgical resection degrees between the two groups (P>0.05). There were 8 patients with 1p/19q co-deletion and 74 patients without 1p/19q co-deletion; no significant differences in above clinical parameters were noted between the two groups. There were no statistically significant differences in the incidences of bone marrow suppression, digestive tract response and fatigue, disease progression rate, or survival rate between patients from TERTp wild type group and TERTp mutation group, and between patients with 1p/19q co-deletion and patients without 1p/19q co-deletion (P>0.05). No significant differences in above clinical parameters, disease progression rate, and survival rate were noted between patients with C228 mutation and C250 mutation (P>0.05). Conclusion TERTp typing and 1p/19q co-deletion status do not have prognostic value in newly-diagnosed MGMT un-methylated/IDH wild-type GBM patients; patients with TERTp mutations have older age than wild-type patients; patients with C250 mutation trend to have higher survival rate than those with C228 mutation.
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Objective To investigate the protective effect and the possible mechanism of intravenous infusion with perfluorocarbon pretreatment and posttreatment on acute lung injury in rats on LPS-induced acute lung injury in rats.Methods A total of 24 Wistar rats were randomly divided into 4 groups:control group(NS group),LPS group,Pre group and Post group.Normal saline was given to NS group as a control.Rats were treated with LPS by intratracheal instillation in LPS group,rats received PFC through femoral vein.prior to LPS instillation in Pre group and rats received PFC through femoral vein after LPS instillation in Post group.NS group were sacrificed at 6 h after being injected with NS,LPS group,Pre group and Post group were sacrificed at 6h after being given LPS.Pathological changes of king,PaO2,lung wet to dry weight ratio (W/D),expression of myeloperoxidase (MPO) and intercellular adhesion molecule-1 (ICAM-1) of lung were assessed.Results Intravenous.infusion with perfluorocarbons increased PaO2,decreased W/D,MPO and ICAM-1 significantly (P＜0.05),and this effect is more remarkable in Pre group.Conclusion Intravenous infusion of PFC significantly protects lung from acute lung injury,especially by pretreatment forms,probably by down-regulate the expression of ICAM-1.
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AIM:To study whether cordycepin (Cordy) plays a role in myocardial protection by regulating the expression of microRNA-455 ( miR-455) and reducing apoptosis induced by endoplasmic reticulum stress in the ischemia -reperfusion (IR) rats.METHODS: SD rats (250 ~300 g) were randomly divided into 3 groups: control group, the chests of the rats were only opened;IR group, the rats were given myocardial ischemia for 30 min, and then reperfusion for 120 min;IR+Cordy group:before IR, the rats were given Cordy (10 mg/kg) through femoral vein injection once a day for one week, and the last injection was given 30 min before ischemia.Automated biochemical analysis was used to detect rat serum activity of LDH and CK-MB.The myocardial endothelial cell ( EC) apoptotic rate was measured by TUNEL , and the ultrastructural changes of the myocardial EC were observed under transmission electron microscope .RT-qPCR was used to detect the expression of miR-455 and the mRNA levels of glucose-regulated protein 78 ( Grp78) and caspase-12 in the myo-cardium.RESULTS:Compared with control group, EC apoptosis in IR group was significantly increased (P<0.05). Compared with IR group , EC apoptosis in IR +Cordy group decreased significantly ( P<0.05 ) .Compared with control group, swelling of mitochondria , irregular membrane , loose wrinkles with vacuoles , disappeared matrix granules , irregular nuclear membrane , chromatin condensation , disappeared nucleoli and even small apoptosis body in the EC were found in IR group.Compared with IR group , the symptoms in IR +Cordy group were greatly improved .Compared with control group, the mRNA expression of Grp78 and caspase-12 as well as the miR-455 level in IR group was increased (P<0.05). Compared with IR group , the mRNA expression of Grp78 and caspase-12, as well as the miR-455 level in IR+Cordy group was decreased (P<0.05).CONCLUSION:Cordycepin attenuates myocardial EC apoptosis , down-regulates the expres-sion of miR-455, and inhibits endoplasmic reticulum stress in the myocardium .
摘要
Objective To establish an AGS cell line that stably expressing miR?126 and to study the effect of miR?126 on the proliferation and metastatic abilities of the AGS cell line in vitro. Methods AGS cells were infected by lentivirus with Lv?has?mir?126. After confirmation by RT?PCR ,CCK?8 and clone formation assays were used to evaluate the effect of miR?126 on AGS cell growth. Transwell migration and invasion assays were used to evaluate the effect of miR?126 on metastasis of AGS cells. Results We verified correct construction of recombinant AGS cells. RT?PCR confirmed mRNA levels of miR?126 existed significantly differences among the recombinant cell lines (P< 0.05). Proliferation assays and clone formation assays did not show a remarkable growth suppression in AGS?mir?126 cell line. However,transwell assay showed a notable acceleration in AGS?mir?126(P< 0.05). Conclusions We successfully constructed recombinant AGS cell line with stably high miR?126 expression level. MiR?126 could facilitate the metastasis of AGS cell in vitro.
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SUMMARY Trauma is a global social problem, with the number of deaths up to 5.8 million all over the world annually.Currently, severe trauma has become the first cause of death in young adults in China. Nowadays, there are many problems in the trauma rescue system, including long pre-hospital transfer pe-riod , several secondary transfers, no information exchange between pre-hospital and in-hospital care, and the poor integrated treatment, which results in the situation that the overall treatment level of severe trau -ma in China is relatively low.In order to solve these problems, we carried out the research and promotion of severe trauma rescue standard, involving completing severe trauma information database , providing lo-cal rescue medical workers with standard training , and building up the information system for the linkage and warning of severe trauma.In addition, we developed and promoted the new standard system for se -vere trauma in 15cities with 124 medical centers.Due to our research, the treatment ability of severe trauma in the pilot areas was enhanced, and the mortality and morbidity of severe trauma were reduced significantly.To sum up, we had got the expected results after implementing the project .
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Objective:To identify the characteristics and risk factors of the refractures after percuta-neous kyphoplasty ( PKP) and percutaneous vertebroplasty ( PVP) .Methods:A retrospective analysis of 148 patients who had undergone PKP or PVP between March 2006 and October 2013 inPeking University People’ s Hospital was conducted.In the study, 29 patients with 42 refractured vertebra and 119 patients without refracture were included.All the patients were observed for a time of (34.4 ±26.8) months. Clinical, imaging and procedure related factors ( gender, age, height, weight, body mass index, the level of the injured vertebra, the time interval between the procedure and the refracture, the level of the refractured vertebra, the bone cement volume injected, performed PKP or PVP,performed unilateral or bilateral, the percentage of anterior vertebral height restoration, the correction of the Cobb angle, cement diffusion, bone mineral density, presence or absence of diabetes mellitus, history of fractures of the whole body, anti-osteoporosis treatment, cement leakage) for each group were analyzed by Cox propor-tional hazards regression analysis.Results:Of all the patients,16 (55.17%, 16/29) had refractures in the adjacent vertebra, and 13 (44.83%, 13/29) had refractures in the nonadjacent vertebra.Refrac-tures within 3 months accounted for 31.03%(9/29) of all the refractures, and within 1 year accounted for 55.17%(16/29).Both older age (P=0.027, HR=1.051, 95%CI=1.006-1.098) and a his-tory of fractures of the whole body (P=0.012, HR=0.386, 95%CI=0.184-0.812) were statistical-ly significant as the independent risk factors for predicting refractures.Others were not associated with re-fractures ( P>0.05) .Conclusion:Older age and a history of fractures of the whole body are the inde-pendent risk factors of the refractures after PKP and PVP.The mechanism of the refractures after PKP and PVP is mainly the natural development of osteoporosis.
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Objective To investigate the potential therapeutic efficacy of lentivirus-mediated artemin (ARTN) gene modified bone marrow mesenchymal stem cells (MSCs) transplantation on the rat model of Parkinson' s disease (PD) and the effects on expression of brain-related proteins.Methods MSCs were isolated and cultured in vitro,transfected by recombinant lentiviral vectors carrying ARTN gene.The PD rat model established by 6-hydroxydopamine (6-OHDA) was randomly divided into 5 groups:Sham group,PD group,MSCs group,MSCs transfected with empty lentiviral vectors transplanted (LV-MSCs)group and MSCs transfected with recombinant lentiviral vectors carrying ARTN gene transplanted (LVARTN-MSCs) group.The MSCs,LV-MSCs and LV-ARTN-MSCs groups were transplanted into the left striatum of each rat model of PD and ethology tests in every group were made with intraperitoneal injection of apomorphine (APO) 2,4,6,8 weeks after transplantation.The expression of tyrosine hydroxylase (TH) protein in substantia nigra (SN) was measured by Western blotting and immunohistochemistry,and immunofluorescence showed ARTN gene modified MSCs expression in rat brain tissue.The levels of dopamine (DA),dihydroxy-phenylacetic acid and homovanillic acid in striatum of each group were detected by high performance liquid chromatography.Results After injection of APO,rotation frequency decreased in LV-ARTN-MSCs group,i.e.(179.33 ± 10.74) circles/30 min vs (235.83 ± 18.95),(203.67 ±11.50) and (206.33 ± 11.86) circles/30 min in PD,MSCs and LV-MSCs groups (q =8.828,P ＜ 0.01;q =3.802,P ＜ 0.05;q =4.219,P ＜ 0.05).The percentage of TH-positive cells in SN after cell transplantation was increased significantly in LV-ARTN-MSCs group (64.05％ ± 5.49％) when compared with PD group (34.18％ ±3.35％),MSCs group (52.59％ ±4.48％) and LV-MSCs group (50.57％ ± 4.41％),respectively (q =13.280,5.135,6.028,all P ＜0.01).At the same time,TH protein in SN after cell transplantation was also increased obviously in LV-ARTN-MSCs group.ARTN gene modified MSCs can survive for at least 6 weeks in the rat brain of PD,mainly concentrated in the transplantation side of striatum.Eight weeks later,the levels of DA in striatum after cell transplantation were elevated significantly in MSCs group (2.34 ± 0.54),LV-MSCs group (2.28 ± 0.45) and LV-ARTN-MSCs group (2.28 ± 0.45)when compared with PD group (0.87 ± 0.07) (q =5.233,P ＜ 0.05;q =5.020,P ＜ 0.01;q =20.190,P ＜ 0.01),and LV-ARTN-MSCs group showed the most significant improvement.Conclusion ARTN gene modified bone marrow MSCs transplanted into the striatum of brain may have therapeutic effects on rat models of PD.
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Objective To discuss the protective effect of Artemin (ARTN) on 6-hydroxydopamine (6-OHDA)-mediated neurotoxic injury,and the alternations of related proteins in Parkinson's disease (PD) model rats.Methods Bone marrow mesenchymal stem cells (MSCs) were isolated and cultured in vitro.After being transfected with recombinant lentiviral vectors carrying A RTN gene,MSCs stably expressed ARTN were chosen (Lv-ARTN-MSCs cells).The SH-SY5Y cells were treated with supematant of Lv-ARTN-MSCs prior to 6-OHDA treatment,and then,cell survival rate was measured by MTT assay and morphologic changes in cultured SH-SY5Y cells were observed by fluorescence microscope (Hochest33258 staining).PD rat models were established and randomly divided into four groups (n=6):PD group,MSCs group,Lv-MSCs group and Lv-ARTN-MSCs group; and sham-operated group (n=6) was also chosen.The PD,MSCs,Lv-MSCs and Lv-ARTN-MSCs groups were transplanted with 5 μL of saline,MSCs (1.0×l05 cell/5 μL),empty virus modified MSCs (1.0×105 cell/5 μL) and ARTN gene modified MSCs (1.0×105 cell/5 μL),respectively,into the left striatum; rats in the sham-operated group were injected with saline when rats in the other groups were received 6-OHDA injection (the same surgical procedures and coordinates).The expressions of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the striatum were measured by Western blotting.Results Western blotting indicated obvious ARTN protein expressions in the Lv-ARTN-MSCs groups.The supematant of Lv-ARTN-MSCs could effectively reduce the apoptosis rate induced by 6-OHDA; as compared with that of SH-SY5Y cells in the 6-OHDA group,the cell survival rate in the Lv-A R TN-MSCs group increased by 13.67％,with significant difference (P＜0.05).Eight weeks after transplantation,the levels of TH and DAT protein in the striatum were elevated significantly in MSCs group,Lv-MSCs group and Lv-ARTN-MSCs group as compared with those in the PD group (P＜0.05),and the Lv-ARTN-MSCs group showed the most significant improvement.Conclusion ARTN which is a secreted protein can protect dopaminergic neuron against 6-OHDA-induced toxicities in Parkinson's disease,and the mechanism might be related to the increased expressions of TH and DAT.
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BACKGROUND:Static compressure effect between the fracture fragments was generated by fixation itself (tension band wire and screw), but dynamic compression effects were generated during flexion. Mechanical strength and stability of patel ar fracture fixation have obvious advantages. However, there are lacks of quantitative comparative studies on static and dynamic compression effects of these fixation methods. OBJECTIVE:To observe strength changes and clinical significance of static and dynamic compression using four fixation techniques. METHODS:Standardized transverse patel ar fracture models were created with fresh cow patel as. The patel as were randomly divided into four groups:fixation was accomplished with modified tension band wiring (wire group);modified tension band with braided cable (cable group);interfragmentary screws (screw group);cannulated screw tension band with wire (cannulated screw group). Before fracture fixation, Fuji pressure-sensitive film was laid among fracture fragments to measure the pressure among fracture fragments after fixation, i.e., static and dynamic compression. Model of each group was measured as fol ows:(1) after fixation, the fixation was removed, and the Fuji pressure-sensitive film was taken out;(2) after fixation, material testing machine was used. Samples underwent a three-point bending test with a 5 000 N load, simulating dynamic compression during knee flexion. Subsequently, Fuji pressure-sensitive film was taken out. Each Fuji pressure-sensitive film was tested using prescale FPD-8010E software. Thus, average pressure among broken bone ends was obtained, and statistical analysis was performed. Static and dynamic compression among broken bone ends was compared in each group. RESULTS AND CONCLUSION:Average static compression was significantly lower in the wire group than in the cable group, screw group and cannulated screw group (P0.05). Dynamic compression was higher than static compression in the wire group (P<0.05). Results verified that compared with modified tension band wire fixation technique, cable or screw could evidently increase static compression among broken bone ends, but simultaneously weaken dynamic compression among broken bone ends.
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Objective To investigate the amplifying results of repairing degenerated distal receptor nerves with normal proximal donor nerves.Methods The study was carried out on 12 adult male SD rats that randomly derided into degeneration group and control group.The distal tibial nerve stumps as receptor nerve were denervated for 8 weeks before cross-sutured with partial freshly transected proximal common peroneal nerve stumps in the degeneration group,while in the control group,freshly transected distal tibial nerve stumps were sutured with partial freshly cut proximal common peroneal nerve stumps.The motor nerve conduction (MCV),muscle rigidity contraction force were measured,the histological examinations were carried out and the numbers of myelinated axons were calculated 3 months after the operation.Results The degeneration group showed a poorer regeneration capacity than the control group (P＜0.05).The MCV were (16.992 ± 3.737) m/s,(23.092 ± 2.788) m/s respectively;The ratio of muscle force were (39.642 ±5.865)% and (71.098± 6.778)% respectively.The regenerated myelinated nerve fiber number were 1718.2± 282.0 and 3340.0 ± 506.5 respectively in degeneration group and control group.The amplifying ratio of the degeneration group and control group were 1.581 ± 0.329 and 3.098 ± 0.642.Conclusion The amplifying results of repairing the degenerated nerves are poorer than repairing the freshly injury distal nerves with the freshly transected proximal donor nerves,the results demonstrate a progressive inability of chronically degeneration of the distal nerve to support axonai regeneration.
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Objective To develop a new method for sterilization in agars culture mediums,i.e.Agars culture mediums is sealed to be sterilized and preserved after resolution.Methods The effect of rudimental Ethylene Epoxide was investigated,and the germiculture results by different preserving methods were analyzed.Air sedimentation bacterium was sampled by different methods to analyze the bacterium capture rate.Results Ethylene Epoxide sterilized agars culture mediums bacterium capture rate was similar with regular agars.Conclusion Ethylene Epoxide sterilized agars culture mediums can be used in sampling of clinical treatment and other outburst incident,reducing the preparation time.
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Hypertension, which has variant of pathogenesis, is one of the most widely spread chronic diseases with serious complications. Antihypertensive drugs can reduce blood pressure and alleviate or reverse cardiovascular remodeling by affecting one or more processes of blood pressure adjusting system. This review focus on targets of antihypertensive drugs associated with Renin angiotensin system, Adrenergic system, Bradykinin Prostacyclin system, Endothelia factor relative system, iron channels and the progresses in the studies of gene therapy for hypertension.
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Background and purpose:The prognosis of high grade gliomas remains poor, and multidisciplinary treatment strategy has been much investigated recently. This study was to explore the efficacy of Temozolomide as first-line treatment combined with radiotherapy and followed by adjuvant chemotherapy for the treatment of newly diagnosed high grade gliomas. Methods:18 patients who had been pathologically proven to be high grade gliomas were enrolled into the study. The patients received 40 Gy/20fractions for the whole brain and followed by 20Gy/10fractions as a boost to tumor bed. All of the patients were given daily oral temozolomide 75mg/ m2 during radiotherapy. 4 weeks after radiotherapy, all of the patients received 6 cycles of Temozolomide, each cycle lasted 5 days with 28 days interval between each cycles. 150 mg/m2 of temozolomide was given for the first cycle for five days,followd by 200 mg/m2 of drug for the rest of the cycles if no significant drug related toxicities were observed. Results:Median follow-up was 12.5 months, 11 cases had either recurrence or progression, 5 of them died from the disease. The median time for disease progression-free survival was 9.8 months (95% CI, 6.1~9.8months), the median time for overall survival was 14 months (95% CI, 8.5 ~ 19.5months), 1-year overall survival rate was 55.6% ,6-month progression-free survival rate was 81.8%. there were no severe temozolomide related toxicities. Conclusion: Concurrent temozolomide with radiotherapy and followed by 6 cycles of temozolomide in the treatment of high grade gliomas had better clinical efficacy, the patients tolerated the strategy well and no severe toxicities were observed.
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AIM To investigate the effects of the novel antihypertensive drug iptakalim hydrochloride on potassium currents in artery smooth muscle cells. METHODS The effects of iptakalim hydrochloride on potassium currents in smooth muscle cells derived from rat pulmonary arteries were observed by using patch clamp technique(whole cell recording) after application of the drug in the bath. RESULTS The potassium current-voltage curves ( I-U curves) of smooth muscle cells derived from normotensive rat pulmonary arteries were up-ward shifted by iptakalim hydrochloride(0.1,1,10 and 100 ?mol?L -1 ). Within 5 minutes after application of the drug, the current amplitude could increase to[(118.6?15.9)%, P