[Objective]To compare retrospectively the clinical outcome in total knee arthroplasty with and without patellar resurfacing.[Method]From October 1994 to October 2004,125 patients(148 knees)with osteoarthritis were enrolled in the clinical trial of total knee arthroplasty,patients received resurfacing(84 knees)or retention patellar(64 knees).Evaluation was done from preoperatively to final followed-up postoperatively.The patients were scored using American HSS Score for knee and the Feller Score for patalla.The data were analyzed using SPSS software.[Result]In resurfacing patella group,the preoperative and final follow-up HSS scores of patients were(39.6?9.8)and(90.9?8.2)points respectively,scores of patailar were(14.4?6.4)and(25.2?4.8),scores of anterior knee pain were(4.6?3.9)and(10.6?4.1).In nonresurfacing patella group,the preoperative and final follow-up HSS scores of patients were(38.8?9.8)and(90.2?8.9)points respectively,scores of patallar were(14.2?6.2)and(25.1?4.8),scores of anterior knee pain were(4.8?3.8)and(10.3?4.1).There were significant differences in both groups preoperatively and postoperatively in anterior knee pain score,but no significant difference between the two groups in anterior knee pain score.[Conclusion]Whether the patellar replaced or not,the most importance for avoiding anterior knee pain postoperatively is the reasonable selection of patient and correct operation.

BACKGROUND: Bone morphogenetic protein 2 and transforming growth factor β are important factors in bone regeneration, increasing the expressions of bone morphogenetic protein 2 and transforming growth factor β can promote the osteogenic differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To construct the lentivirus vector carrying bone morphogenetic protein 2 and transforming growth factor β3, and to observe the expression of lentivirus vector in bone marrow mesenchymal stem cells. METHODS: The recombinant lentiviral vectors carrying transforming growth factor β3, bone morphogenetic protein 2 and green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect the passage 3 rabbit bone marrow mesenchymal stem cells in vitro cultured (transfection group). The bone marrow mesenchymal stem cells transfected with single gene lentivirals (single gene transfection group) carrying transforming growth factor β3 and bone morphogenetic protein 2 or single lentivirals were as control (control group). At 1 week after trasfection, the total RNA and protein were extracted from each group for detection. RESULTS AND CONCLUSION: The green fluorescence bone marrow mesenchymal stem cells transfected with transforming growth factor β3 and bone morphogenetic protein 2 gene for 3 days could be observed under fluorescence microscope, and the transfection efficiency was over 90%. Reverse transcription-PCR and Western blot results showed the mRNA and protein expressions of transforming growth factor β3 and bone morphogenetic protein 2 in the transfection group were higher than those in the single gene transfection group and the control group. The results indicate that lentivirus can successful y transfect transforming growth factor β3 and bone morphogenetic protein 2 into the bone marrow msenchymal stem cells and achieve its high expression, and these two genes have the synergistic effect of promoting expression.

BACKGROUND: The existed repair method for cartilage defects has shortcomings of insufficient repairing tissue numbers, poor biomechanical properties, as well as donor site complication. Thus it is deficient to repair large-sized osteochondral defects using one method.OBJECTIVE: To investigate the therapeutic effect of tissue engineering modified Mosaicplasty on repairing large-sized osteochondral defects.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at the Center Laboratory of Qingdao University Medical College from January to September 2009.MATERIALS: The hircine bone mesenchymal stem cells (BMSCs) were in vitro cultured, and resuspended with algin solution to obtain BMSCs-calcium alginate gel.METHODS: Totally 12 goats were prepared for osteochondral defects models and were divided into 3 groups.BMSCs-Mosaicplasty group, BMSCs compound with injectable alginate calcium gel was then applied to fill the "dead space" after Mosaicplasty. In the Mosaicplasty group, the defects were repaired by Mosaicplasty. There was no treatment in the control group.From 4 to 16 weeks postoperatively, the animals were sacrificed and the in gross and under electromicroscopy.MAIN OUTCOME MEASURES: ①Gross observation: the joint was exposed to observe the repair effect at weeks 4, 8,16 after operation. ②Histological examination: specimens were harvested at 16 weeks after operation and observed by haematoxylin-eosin staining, toluidine blue staining under light microscopy. ③Transmission electron microscope was used at 16 weeks after operation.RESULTS: The transplanted subchondral bone and superficial cartilage was integrated hardly with each other or with recipient sites in tissue engineering modified Mosaicplasty groups at 16 weeks after operation. The quality and appearance of the transplanted and regenerated cartilage was similar to normal hyaline cartilage. Under microscopy, the regenerated cartilage was integrated with neighbor tightly in regular arrange. ECM distributed evenly and deeply stained by alcian blue. There was no obviously repaired in the control group.CONCLUSION: Tissue engineering can ameliorate the outcome of Mosaicplasty to repair the osteochondral defects.

OBJECTIVE To explore the role and mechanism of dental pulp stem cells(DPSCs)in repairing hypoxic injury to rats pulmonary microvascular endothelial cells(PMVECs).METHODS ①PMVECs were treated with cobalt chloride at 0,10,25,50 and 100 μmol·L-1 for 72 h.CCK-8 was used to detect the cell viability,and the protein levels of hypoxia-inducible factor 1α(HIF-1α),zona occludens small-band protein 1(ZO-1),and occludin(OCLN)were detected by Western blotting.②There was a cell control group,model group,and model+DPSCs group,and the levels of reactive oxygen species(ROS)was detected by immunofluorescence staining after at 24 and 48 h of action.The levels of ZO-1 and OCLN proteins were detected by Western blotting.③ A cell control group,model group,model+DPSC group and model+DPSC cell knockdown superoxide dismutase 1(SOD1)group were set up.The mRNA level of SOD1 was detected by real-time fluorescence quantitative PCR 24 and 48 h later,while the protein levels of ZO-1 and OCLN were detected by Western blotting.RESULTS ① Com-pared with the cell control group,72 h of cobalt chloride 100 μmol·L-1 treatment of PMVECs resulted in a cell survival rate above 80%,a significant increase in the level of HIF-1α protein(P<0.05),a signifi-cant decrease in the levels of ZO-1 and OCLN proteins(P<0.01),and establishment of a model of hypoxic injury in PMVECs.② Compared with the cell control group,the ROS level was significantly higher in the model group(P<0.01).Compared with the model group,the ROS level was significantly lower in the model+DPSCs group(P<0.01),while the levels of ZO-1 and OCLN proteins were signifi-cantly higher in the model+DPSCs group(P<0.05).③ Compared with the DPSC group,ZO-1 and OCLN expressions were significantly decreased after knockdown of SOD1 in DPSCs(P<0.05,P<0.01).CONCLUSIONS DPSCs can repair hypoxic injury to PMVECs,and the anti-oxidative stress capacity of DPSCs plays an important role in hypoxic injury repair of PMVECs.

Objective To explore the effect of neostigmine in treatment of severe acute pancreatitis (SAP) combined with intra-abdominal hypertension (IAH).Methods 42 patients diagnosed as SAP combined with IAH and meeting standard were collected from Aug.2012 to Jul.2016 in our hospital.They were randomly divided into two groups:observation group and the control group.General information of the two groups was comparable.Patients of the control group received conventional treatment methods such as fasting,gastrointestinal decompression,nutritional support,antispasmodic,analgesia,intravenous infusion of omeprazole and octreotide.Patients of the observation group received intramuscular injection of neostigmine methylsulfate on the basis of the control group for seven days.The intra-abdominal pressure,recovery time of bowel sounds,the first exhaust and defecation time,remission time of abdominal distention,modified Marshall score,APACHE-Ⅱ score,SIRS score,total hospitalization time and total hospitalization expenses,MODS rate,operation rate and mortality of the two groups were recorded during the treatment.Results The intra-abdominal pressure of the observation group was (14.25±1.03) mmHg,(13.52 ±1.23) mmHg,(12.73±1.14)mmHg respectively,lower than that of the control group (15.14± 1.12) mmHg,(14.60± 1.11) mmHg,(13.84±1.08) mmHg at the 3rd,4th,and 5th day after treat ment (P＜0.05).For the observation group,the recovery time of bowel sounds was (7.24±1.35) d,the first exhaust and defecation time was (11.33±1.51) d,(8.36±1.63) d,remission time of abdominal distention was (13.62±2.26) d,lower than those of the control group ((9.56±1.17)d,(13.42±1.26)d,(10.45±1.54)d,(16.75±3.05)d) (P＜0.05).The recovery time of bowel sounds,the first exhaust and defecation time,remission time of abdominal distention of the observation group were lower than those of the control group (P＜0.05).The modified Marshall score,APACHE-]Ⅱ score,SIRS score of the two groups had no significant difference (P＞0.05).The total hospitalizationtime and total hospitalization expenses of the two groups had no significant difference (P＞0.05).MODS rate,operation rate and mortality of the two groups had no significant difference (P＞0.05).Conclusions Neostigmine can significantly reduce intra-abdominal pressure of patients suffering from SAP combined with IAH,improve the symptoms of paralytic ileus,promoting exhaust and defecation.However,it has limited effects on prognosis of patients suffering from SAP combined with IAH.

BACKGROUND:Stable intertrochanteric fractures can be treated by closed reduction and internal fixation,but there is no absolute advantage for unstable intertrochanteric fractures with osteoporosis. OBJECTIVE:To investigate the efficacy of arthroplasty in the treatment of unstable intertrochanteric fractures in the elderly with osteoporosis by comparing the indexes related to closed reduction internal fixation and arthroplasty. METHODS:Clinical data of 102 elderly patients with unstable intertrochanteric fractures of the femur treated in Affiliated Hospital of Qingdao University from January 2017 to January 2020 were retrospectively analyzed.Patients were divided into two groups according to the surgical method.In the Gamma3 group,62 cases received Gamma3 internal fixation system.In the joint replacement group,40 cases received an artificial femoral head replacement or total hip replacement.Surgical information,hospitalization,hip function,and postoperative complications were compared between the two groups. RESULTS AND CONCLUSION:(1)There were statistical differences between the Gamma3 group and the joint replacement group in weight-bearing time(P＜0.001),hospital stay(P＜0.05),intraoperative bleeding(P＜0.001),and length of surgery(P＜0.001).The mean weight-bearing time and hospital stay were shorter in the joint replacement group than in the Gamma3 group.Intraoperative bleeding and duration of surgery were better in the Gamma3 group than in the joint replacement group.(2)There was no significant difference in Harris hip score,subitem centesimal hip score,and postoperative complications 12 months after surgery in both groups(P=0.526,0.788,0.228).(3)It is indicated that arthroplasty has achieved better outcomes in the treatment of elderly unstable intertrochanteric fractures combined with osteoporosis,enabling early weight bearing and functional exercise.Careful selection of the appropriate patient and prosthesis type for arthroplasty will result in greater patient benefit.

Objective To explore the effects of the same dose of fractionated radiation and single radiation on radiation-induced heart damage in mice.Methods Twenty-one wild-type C57BL/6J mice were randomly divided into the normal group,fractionated radiation group and single radiation group.18 Gy X-ray,via fractionated(3 Gy/time,6 times)radiation or single radiation,was used to establish a radiation-induced heart damage model.The concentrations of myocardial enzyme damage markers(creatinekinase(CK),creatinekinase-MB(CK-MB),lactate dehydrogenase(LDH)and LDH1)and peripheral serum ions(K+,Ca2+,Fe2+and Cl-)were detected by an automatic biochemical analyzer at day 7 and 28 after radiation.Ultrasound was used to detect and analyze the cardiac function of mice at day 28 after radiation,including the left ventricular ejection fraction(EF),left ventricular fractional shortening(FS),left ventricular end-diastolic volume(LVEDV),left ventricular mass(LV mass)and left ventricular end-systolic volume(LVESV).The opening of the mitochondrial permeability transition pore(mPTP)and changes of mitochondrial membrane potential of myocardial cells were observed using a laser confocal microscope.The ultrastructure of myocardia was observed under a transmission electron microscope(TEM)and cardiac fibrosis was checked by Masson staining.The atherosclerosis of the aorta was examined by gross oil red staining.Real-time quantitative PCR(RT-qPCR)and Western blotting were performed to detect the expressions ofapoptosis-related genes and proteins,B cell lymphoma-2-associated X protein(BAX)and casepase-3.Results Seven days after 18Gy X-ray irradiation,the expression levels of CK,CK-MB,LDH and LDH1 in the single radiation group increased significantly or trended up,while only CK and LDH1 in the fractionated irradiation group continued to increase.Twenty-eight days after radiation,the expression levels of 4 enzymes in myocardial zymogram were increased by both radiation methods.Seven and twenty-eight days after radiation,the concentrations of serum ions K+,Fe2+,Ca2+and Cl-were significantly decreased by both radiation methods that could lead to the decrease of EF and FS,and the increase of LV mass,LVEDV and LVESV.Single radiation made more difference to EF and FS,and the difference between the two groups was statistically significant.Both methods could decrease the mPTP and mitochondrial membrane potential,especially single radiation,and there was significant difference between the two groups.The results of TEM showed that the mitochondrial cristae of myocardial cells decreased and vacuolated,and the myocardial fiber bundles became thicker after X-ray radiation.Masson staining showed that collagen fibers were deposited in the heart tissue ofmice after X-ray irradiation,particularly in the single radiation group.Gross oil red staining ofthe aorta showed that both methods could damage the aorta of mice,and the area of atherosclerotic plaques in the single radiation group was larger,which was statistically different from that of the fractionated radiation group.The results of RT-qPCR and Western blotting showed that X-ray radiation could increase the expression levels of apoptosis-related BAX and caspase-3 in cardiac tissue,especially in the single radiation group.The mRNAexpressions of BAX and caspase-3 increased more significantly in the single radiation group.Conclusion Both fractionated radiation and single radiation at the same dose can cause heart damage,so they can be used to establish a radiation-induced heart damage model of mice.Single radiation can cause more significant damage to the heart.Different modeling methods can be selected as required.

OBJECTIVE To observe whether mitochondria can be transferred from mesenchymal stem cells(MSCs)to irradiated cells by establishing a mitochondrial fluorescent reporting system.METHODS The lentiviral vector pSIN-EF1α-COX8A-DsRed2(named COX8A-DsRed2)that might guide the expres-sion of red fluorescence protein in the membrane of mitochondria was constructed.A lentivirus(named Lv-COX8A-DsRed2)was prepared in 293T cell line.Dental pulp stem cells(DPSCs)(named DPSC-COX8A-DsRed2)was infected with Lv-COX8A-DsRed2.The intracellular expression of the red fluores-cence protein in DPSC was observed under fluorescence microcopy.The mitochondrial localization of the expressed red fluorescent probe in DPSC-COX8A-DsRed2 was confirmed according to TOMM20 immunostaining and MitoTracker Green staining results,which could specifically label mitochondria.The IEC-6 cells that received 10 Gy X-ray radiation were used as an injured cell model.The co-culture system was established by supplementing DPSC-COX8A-DsRed2 into the culture plate with the irradi-ated IEC-6 labelled by CFSE for 24 h.RESULTS The imaging results of fluorescent microcopy obser-vation showed that DPSC-COX8A-DsRed2 expressed the mitochondrial fluorescent reporting system,which was co-located with TOMM20 protein and Mito Tracker Green.The imaging results of confocal fluorescence microcopy showed that the mitochondria with red fluorescent protein were transferred from DPSC-COX8A-DsRed2 to the irradiated IEC-6 cells,suggesting that the established mitochondrial fluorescent reporting system could indicate mitochondrial transfer from donor cells to injured ones.CONCLUSION DPSC-COX8A-DsRed2 stably expressing the mitochondrial fluorescent reporting system is established,which can be used to track mitochondrial transfer.

Objective To evaluate the protective effect of dental pulp stem cells(DPSCs)against high-altitude pulmonary edema(HAPE)and explore the mechanism.Methods Twenty-one Sprague-Dawley rats were randomly divided into the normal control group(Control),high altitude pulmonary edema group(HAPE)and DPSC prevention group(DPSC).Three days and one day before the hypobaric and hypoxia environment was simulated,DPSCs were administrated via the tail vein in the DPSC group,and phosphate buffered saline(PBS)of the same volume was injected in the control group and HAPE group.After that,rats in the HAPE group and DPSC group were placed in a simulated hypobaric and hypoxic cabin to simulate the environment at an altitude of 6000 m before an HAPE model was established.The rats were sacrificed 3 days later,and the histopathological changes of lung tissue were observed via hematoxylin-eosine(HE)staining.The dry/wet weight of lung tissues was recorded.The concentrations of total protein in lung homogenate were detected using the BCA method.The expression of aquaporin 1(APQ-1)in lung tissue was detected by real-time reverse transcription polymerase chain reaction(RT-PCR)and Western blotting.Colorimetry and enzyme linked immunosorbent assay(ELISA)were used to detect the concentrations of plasma vasoactive substances.The expression of inflammatory factors in serum and lung tissue was detected by ELISA and RT-PCR respectively.Results After three days of treatment in a simulated hypobaric and hypoxic environment,the lung tissue of the HAPE group showed infiltration of inflammatory cells,alveolar wall thickening,alveolar septum thickening and increases of exudation.DPSC prevention could significantly reduce the histopathological damage to the lung.Lung water content(LWC)and total protein content of lung homogenate were significantly increased in the HAPE group but obviously reduced in the DPSC prevention group,and the difference between the two groups was statistically significant(P＜0.01).The protein and mRNA expression levels of AQP-1 in lung tissue of the HAPE group were significantly decreased,but prevention via DPSCs could enhance the expression of AQP-1 in lung tissue.In the HAPE group,the concentrations of nitrous oxide(NO)and prostacyclin(PGI2)in plasma were significantly decreased,but prevention via DPSCs could significantly increase the expression of plasma vasoactive substances.The expressions of inflammatory factors interleukin-1 α(IL-1α)and tumor necrosis factor-α(TNF-α)in serum and lung tissue of the HAPE group increased significantly,but decreased in the DPSC prevention group.Conclusion DPSCs can help prevent high-altitude pulmonary edema in rats by promoting the expression of aquaporin,increasing the content of vasodilators in plasma and reducing the expression of inflammatory factors in serum and lung tissue.