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Objective:To evaluate the effects of different densities of rat cardiac fibroblasts (RCF) subjected to hypothermic hypoxia-reoxygenation on cardiomyocyte injury and intercellular coupling.Methods:RCF was cultured in vitro and divided into 3 groups ( n=12 each) using a random number table method: RCF density 0.5×10 5 cells/ml group (T 0.5 group), RCF density 1.0×10 5 cells/ml group (T 1.0 group), and RCF density 2.0×10 5 cells/ml group (T 2.0 group). The three groups were placed in an anoxic device, into which 95% N 2 + 5% CO 2 was continuously blown at the speed of 5 L/min for 15 min, and then placed in a 4 ℃ refrigerator for 1 h for low temperature treatment.After completion of culture, cells were placed in a incubator containing 95% air + 5% CO 2 at 37 ℃ for 4 h of reoxygenation.After the end of culture, RCF in three groups were indirectly co-cultured with cardiomyocytes of the same density (1.0×10 5 cells/ml) in a Transwell chamber for 16 h, cardiomyocytes were seeded in the lower chamber of Transwell, and RCF were seeded in the upper chamber of Transwell.After the end of co-culture, cardiomyocytes were collected for determination of the cell viability (by CCK8 method), apoptosis rate (by flow cytometry), expression of connexin 43 (Cx43) mRNA (by real-time fluorescence quantitative polymerase chain reaction), and expression of Cx43 and phosphorylated Cx43 (p-Cx43) (by Western blot). Results:Compared with T 0.5 group, the cell viability, apoptosis rate and expression of Cx43, p-Cx43 and Cx43 mRNA were significantly decreased in T 1.0 and T 2.0 groups ( P<0.01). Compared with T 1.0 group, the cell viability, apoptosis rate and expression of Cx43 and p-Cx43 were significantly decreased ( P<0.01), and no significant change was found in expression of Cx43 mRNA in cardiomyocytes in T 2.0 group ( P>0.05). Conclusions:RCF subjected to hypothermic hypoxia-reoxygenation induces cardiomyocyte injury in a density-dependent manner in a certain range, and the mechanism may be related to down-regulation of the expression of Cx43 and reduction of the activity of Cx43.
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Objective:To evaluate the effect of rat cardiac fibroblasts (RCF) on the expression of connexin43 (Cx43) in H9c2 cells during hypothermic hypoxia/reoxygenation.Methods:H9c2 cells cultured in vitro were divided into 4 groups ( n=12 each) using the random number table method: control group (group C), hypothermic hypoxia/reoxygenation group (group HHR), RCF co-culture group (group Co) and RCF co-culture plus hypothermic hypoxia/reoxygenation group (group Co+ HHR). Group C was incubated at 37℃ in 5% CO 2 + 95% air for 5 h. Group HHR was incubated at 4 ℃ in 5% CO 2 + 95% N 2 for 1 h and then at 37 ℃ in 5% CO 2 + 95% air for 4 h. In group Co and group Co+ HHR, H9c2 cells 0.3×10 5 cells/well were inoculated in the lower chamber and RCF 0.6×10 5 cells/well in the the upper chamber of a transwell ? culture dish.Group Co was incubated at 37 ℃ in 5% CO 2 + 95% air for 5 h. Group Co+ HHR was incubated at 4℃ in 95% N 2 + 5% CO 2 for 1 h, and then incubated at 37 ℃ in 5% CO 2 + 95% air for 4 h. The mortality rate of H9c2 cells was measured by trypan blue staining, the expression of Cx43 and extracellular signal-regulated protein kinases 1/2 (ERK1/2) by immunofluorescence, and the expression of Cx43, phosphorylated Cx43, ERK1/2 and phosphorylated ERK1/2 by Western blot. Results:Compared with group C, the mortality rate of H9c2 cells was significantly increased, the expression and phosphorylation of Cx43 were decreased, and the expression and phosphorylation of ERK1/2 were increased in group HHR ( P<0.05), and no significant change was found in the mortality rate of H9c2 cells or expression and phosphorylation of Cx43 and ERK1/2 in group Co ( P>0.05). Compared with group Co, the mortality rate of H9c2 cells was significantly increased, and the expression and phosphorylation of Cx43 and ERK1/2 were decreased in group Co+ HHR ( P<0.05). Compared with group HHR, the mortality rate of H9c2 cells was significantly increased, and the expression and phosphorylation of Cx43 and ERK1/2 were decreased in group Co+ HHR ( P<0.05). Conclusions:RCFs can decrease the expression and activity of Cx43 in H9c2 cells during hypothermic hypoxia/reoxygenation, and the mechanism may be related to the down-regulation of ERK1/2 expression and inhibition of ERK1/2 activity.
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Macrophages play an important role in maintaining homeostasis of the body, and they are also one of the most abundant immune cells in the tumor microenvironment (TME). These macrophages are often called tumor-associated macrophages (TAMs), which play an important role in the development of tumor and are an important target for tumor therapy. Studies have shown that tumor growth and metastasis can be inhibited by regulating the function of macrophages, but the therapeutic efficacy was often hampered by the poor performance of the drugs such as lack of targeting, poor solubility, low bioavailability, and severe side effects. After introduction of the background of macrophage and tumor therapy, this review focuses on the research progress of nano-drug delivery systems in the modulation of the function of macrophages to enhance tumor immunotherapy. Nano-drug delivery systems are diverse in structures and functions, and can regulate macrophage functions through a variety of mechanisms. Four important aspects of macrophage modulation, which included TAMs depletion, repolarization of TAMs, promoted phagocytosis of TAMs, and combinational modulation of TAMs were summarized. Each strategy together with typical examples was reviewed and future directions in this field were also prospected.
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Objective:To evaluate the effects of exosomes derived from cardiac fibroblasts pretreated with sevoflurane on ventricular electrical conduction in isolated rat hearts subjected to hypothermic ischemia-reperfusion (I/R) using the multi-electrode array mapping technique.Methods:Primary cardiac fibroblasts were extracted by differential adhesion in SPF Sprague-Dawley rats of either sex.Cardiac fibroblasts of passage 2-4 were treated with 2.5% sevoflurane for 1 h, and then cultured for 24-48 h to extract exosome.SPF healthy male Sprague-Dawley rats, aged 2-3 months, weighing 280-320 g, were divided into 3 groups ( n=8 each) using a random number table method: control group (group C), I/R group and sevoflurane-pretreated cardiac fibroblast-derived exosome+ IR group (group S+ IR). Hearts were perfused for 110-min equilibration in group C. After 20 min of equilibration, the perfusion was suspended for 60 min (global ischemia) followed by 30 min of reperfusion in IR and S+ IR groups.Exosomes 1 ml (200 μg) derived from cardiac fibroblasts pretreated with sevoflurane were injected through the tail vein at 48 h before surgery in group S+ IR, and the equal volume of normal saline was injected instead in C and IR groups.The cardiac conduction velocity (CV), conduction absolute inhomogeneity (P 5-95) and inhomogeneity index (P 5-95/P 50) were obtained at 20 min of equilibration (T 0) and 15 and 30 min of reperfusion (T 1, 2) using the microelectrode array attaching to the left ventricular surface of the isolated heart. Results:Compared with group C, CV was significantly decreased and P 5-95 and P 5-95/P 50 were increased at T 1 ( P<0.05), and no significant change was found at T 2 in group S+ IR ( P>0.05), and CV was significantly decreased and P 5-95 and P 5-95/P 50 were increased at T 1, 2 in group IR ( P<0.05). Compared with group IR, CV was significantly increased and P 5-95 and P 5-95/P 50 were decreased at T 1, 2 in group S+ IR ( P<0.05). Conclusions:Exosomes derived from cardiac fibroblasts pretreated with sevoflurane can improve ventricular electrical conduction in isolated rat hearts subjected to hypothermic I/R.
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Objective:To evaluate the effects of different density rat fibroblasts on the expression of conjunctin 43 (Cx43) in cardiomyocytes and cell viability.Methods:Cardiomyocytes and fibroblasts were co-cultured using Transwell, cardiomyocytes were inoculated into the lower chamber of Transwell and fibroblasts into the upper chamber of Transwell.The cells were divided into 3 groups ( n=12 each) by a random number table method: fibroblast density 0.5×10 5 cells/ml group (group C 0.5), fibroblast density 1×10 5 cells/ml group (group C 1), and fibroblast density 2×10 5 cells/ml group (group C 2), with the density of cardiomyocytes 1×10 5 cells/ml in three groups.Cardiomyocytes and fibroblasts were co-cultured for 20 h in three groups.Cardiomyocytes were collected after co-culture for determination of cell viability (by CCK8 method), apoptosis rate (by flow cytometry), and expression of Cx43 mRNA (by quantitative real-time polymerase chain reaction) and expression of Cx43 and phosphorylated Cx43 (p-Cx43) (by Western blot). Results:There was no significant difference in the apoptosis rate of cardiomyocytes among the three groups ( P>0.05). Compared with group C 0.5, the expression of Cx43 protein and mRNA and p-Cx43 was significantly up-regulated in group C 1, the cardiomyocyte viability was significantly increased, and the expression of Cx43 protein and mRNA and p-Cx43 was up-regulated in group C 2 ( P<0.05). Compared with group C 1, the cardiomyocyte viability was significantly increased, and the expression of Cx43 protein and mRNA and p-Cx43 was up-regulated in group C 2 ( P<0.05). Conclusion:Rat fibroblasts up-regulate the expression of Cx43 and enhance the activity of Cx43 in cardiomyocytes and enhance cell viability in a density-dependent manner in a certain range.
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OBJECTIVE: To investigate the value of HE4,CA125 and ovarian malignant tumor risk prediction models(ROMA)in the diagnosis of ovarian epithelial malignant tumors.METHODS: The clinical and pathological data of 247 patients with ovarian tumors(EOC 139 cases,BOT 18 cases,and benign ovarian tumor 90 cases)and 39 patients with uterine fibroids in Liaoning Tumor Hospital from September 2016 to August 2017 were retrospectively analyzed. The levels of serum CA125 and HE4 were measured before operation. The ROMA values were calculated and the relationship between CA125,HE4,ROMA values and clinical pathological parameters were analyzed. The diagnostic evaluation index was calculated,the receiver operating characteristic(ROC)curve was drawn,and the AUC value was also calculated.RESULTS: The positive rate of HE4 in the ovarian epithelial malignant tumor group was significantly higher than that in other groups before and after menopause,the difference being statistically significant(P0.05).There was no significant statistical difference in the positive rate of ROMA before or after menopause(P>0.05).The sensitivity of CA125 was higher than that of HE4 and ROMA. Specificity of HE4 was higher than that of CA125 and ROMA.Correct diagnosis index of ROMA was higher than that of HE4 and CA125.CONCLUSION: For the diagnosis of ovarian malignant epithelial tumors,the combined detection of serum HE4 and CA125 and ROMA model is superior to the individual detection of HE4 and CA125.
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Objective to compare and contrast the diagnosis results on 2D and 3D Ultrasonogrpahy against MRI. Method A 2D Ultrasonography was applied during a conventional prenatal sonography checking with a 3D Sonography assessment subsequently conducted to follow up on 49 fetus suspected of brain malformation.Furthermore, a MRI scan was taken within 24 hours after the 3D Sonography checking in our hospital as a final test. Data collections from all three assessments were completed, and an analysis of the comparisons of these three methods were done. Results Among these 49 fetus with confirmed or suspected brain malformation, there were two cases of misdiagnoses of Dandy-Walker Malformation assessed by 2D sonography, with a misdiagnosis rate of 40％, (P ＜ 0.05) indicating a statistically significant result; misdiagnosis rate of Fetal Ventriculomegaly and Isolated Broadening Posterior Fossa were calculated as 26.7％ and 33.3％ respectively (P ＜0.05), a statistically significant result. Overall, there were two cases with Cerebellum Malformation, from in which one case was identified by MRI, and the other one was misdiagnosed, with a misdiagnosis rate of 50.0％.In total, there were 2 cases of Holoprosencephaly, in which one was identified by Prenatal MRI, and the other was misdiagnosed (P ＜ 0.05), a statistically significant result. Conclusions All three assessments of 2D ultrasonography, 3D ultrasonography and MRI have their own advantages and disadvantages. In short, 2D Sonography is suggested to be applied for screening out cases with brain malformation, together with 3D Sonography as a complementary assessment. MRI can also be an effective and significant complement for sonography in completing and readdressing the final ultrasonic results.
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On account of the problem that traditional C-arm X-ray apparatus cannot provide precise route guidance for minimally invasive surgeries, we designed and developed a laser positioning and navigation system based on C-arm X-ray apparatus, which can achieve precise positioning function and reduce the exposure of doctors and patients to radiation in minimally invasive surgeries under the linear guidance of a laser beam. Furthermore, this system can enhance the refinement level of surgical operation in minimally invasive surgeries.
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Humans , Cone-Beam Computed Tomography , Minimally Invasive Surgical Procedures , Radiography , Surgery, Computer-Assisted , Tomography, X-Ray Computed , X-RaysABSTRACT
This paper summarized the nursing experience of one infant with right lower limb ischemia caused by amniotic band syndrome. The nursing key points included:careful observation of the infant condition,exposing the wound,raising the affected limb,wound disinfection,and removing necrotic tissues at the surface skin,applying topical drugs for promoting tissue growth and antibiotics,selecting appropriate dressing to provide suitable environment for tissue regeneration;debridement of inguinal skin,dressing change,assessing pain and providing corresponding care during debridement;infection prevention. After 29 days of treatment and careful nursing care,the skin damage was repaired on the right lower limb,swelling subsided,the necrotic tissues at right inguinal region were completely cleared,granulation tissue was formed,and the infant was discharged smoothly.
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To evaluate the clinical efficacy and safety of Wenxin Granule combined with metoprolol for treatment of ventricular premature. We searched CNKI, Wanfang Database, VIP, Jiangsu Engineering Technology Literature Information Center, CCRCT, Medline, and Sciencedirect from 1995 to 2014. Meta analysis was performed using Rev Man 5.2 software. Twenty trials were involved, including 1 568 cases. The results showed that there was significant difference in the clinical symptom effectiveness of Wenxin Granule combined with metoprolol and metoprolol alone in the treatment of ventricular premature [RR=1.26, 95% CI (1.17, 1.36), P<0.00001], as with the clinical effectiveness [RR=1.22, 95% CI (1.16, 1.29), P<0.000 01] and the incidence of adverse drug reactions [RR=0.70, 95% CI (0.50, 0.98), P=0.04]. A subgroup analysis was done in the view of the severe adverse reaction. The results showed that there was significant difference in the incidence of atrioventricular block between the two groups [RR=0.20, 95% CI (0.04, 0.89), P=0.04], while no significant difference was found in the incidence of bradycardia [RR=0.54, 95% CI (0.26, 1.09), P=0.08]. The results of sensitivity analysis of final indicators were consistent with the fundamental analysis except the incidence of adverse drug reactions. Comparing with metoprolol alone, Wenxin Granule combined with metoprolol for treatment of ventricular premature can effectively improve the symptoms of patients, and significantly diminish the ventricular premature, while there is no significant difference in the incidence of total adverse drug reactions and bradycardia between the two groups. Also the difference in the incidence of atrioventricular block is with marginal significance. So the safety between the two groups may need further study after more samples be tested.
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Objective To review the experiences of diagnosis and treatment of pneumatosis cystoides intestinalis (PCI), and study the clinical characteristics of the disease in order to improve the diagnosis and treatment. Methods Clinical data from 20 patients with endoscopically confirmed PCI were retrospectively analyzed. They were admitted to the Chinese PLA General Hospital from June 1995 to June 2015. Results Among the patients 16 of them were male, and the other four were female. The main clinical manifestations were abdominal distention, diarrhea, abdominal pain and mucous bloody stool. The diagnosis relied mainly on colonoscopy and pathological examination. Laparoscopy assisted colorectal cancer resection was performed in 1 patient, laparostomy and repair of sigmoid colon perforation in 1, endoscopic treatment in 5 cases, drug administration and hyperbaric oxygen therapy in 2, drug treatment alone in 7, and no treatment in 4. Conclusions The final diagnosis depends on endoscopic findings. No treatment is recommended to patients with no symptoms. The management of patients with PCI includes antibiotics, oxygen therapy, endoscopic therapy, surgery, and appropriate therapy related to the underlying cause of PCI. The prognosis is good.
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<p><b>OBJECTIVE</b>To analyze the infection of human parvovirus B19, human bocavirus (HBoV) and human parvovirus 4 (PARV4) in blood samples among patients with liver disease in Nanjing by molecular detection.</p><p><b>METHODS</b>Nested PCR assays were designed and validated to detect B19, HBoV and PARV4, respectively. The assays were used to screen three parvoviruses in blood samples from 95 patients with different liver disease in Nanjing. The parvovirus infection was analyzed statistically.</p><p><b>RESULTS</b>The detection limits were 10 copies of genomic DNA equivalents per reaction for each assays and the good specificity were observed. The frequency of B19 and HBoV were 2/95 (2.1%) and 9/95 (9.5%) in blood samples respectively. No PARV4 was detected. HBoV was detected in 3/5 patients with drug-induced hepatitis.</p><p><b>CONCLUSION</b>Both B19 and HBoV infection were detected in blood from patients with liver disease.</p>
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Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Coinfection , Virology , Human bocavirus , Liver Diseases , Virology , Parvovirus , Parvovirus B19, Human , Polymerase Chain Reaction , Viremia , VirologyABSTRACT
<p><b>BACKGROUND</b>Cyclosporine is effective in treating nephrotic syndrome (NS) with idiopathic membranous nephropathy (IMN) in adults. But high relapse rate remains a major concern. The way to manipulate cyclosporine is inconclusive. The aim of this study was to introduce the way how to titrate the cyclosporine to maintain complete remission without relapse.</p><p><b>METHODS</b>Patients with biopsy-proven IMN with NS treated with cyclosporine for at least 1 month from 1996 to 2011 at Peking Union Medical College Hospital were reviewed.</p><p><b>RESULTS</b>Mean age of the 51 eligible patients was 52 years, with 39 men. Mean proteinuria was (7.47 ± 3.14) g/d, serum albumin (24.50 ± 6.29) g/L, and serum creatinine (82.62 ± 21.18) mmol/L. Cyclosporine was commenced at a mean dose of (3.46 ± 0.63) mg×kg(-1)×d(-1). Oral prednisone (0.40 ± 0.29) mg×kg(-1)×d(-1) was given concomitantly in 38 patients. Cyclosporine was administered for a median of 16 months (range 1 - 93 months) and stopped in non-responders by month six. By month 3 (n = 47), the number in complete remission (CR) and partial remission (PR) was 3 and 24, which shifted to 12 and 17 by month 6 (n = 41). Male gender, heavy proteinuria, low serum albumin level, and high serum creatinine level were significant determinants in poor response by month six (P < 0.05 in all variables compared with responders). There was a significant reversible serum creatinine increase within 25% during month 3 to 12 (P < 0.05 in all variables compared with baseline value). Eleven patients maintained cyclosporine for more than 24 months with a cyclosporine dose of (1.04 ± 1.06) mg×kg(-1)×d(-1). Nine patients were in CR. Renal function, systolic and diastolic blood pressure remained stable. Renal impairment (> 30% rise of serum creatinine), secondary infection, hypertension, gingival hyperplasia and liver impairment occurred in 6, 4, 10, 4, and 1 patients, respectively.</p><p><b>CONCLUSIONS</b>The observation time for cyclosporine to effectively induce CR of NS in IMN adults should be at least six months. Long-term and low-dose of cyclosporine therapy is safe and effective to maintain CR in those responders.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Cyclosporine , Therapeutic Uses , Glomerulonephritis, Membranous , Drug Therapy , Immunosuppressive Agents , Therapeutic Uses , Nephrotic Syndrome , Drug Therapy , Retrospective Studies , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of pricking blood at Neiyingxiang (EX-HN 9) on the intraocular pressure of patients of primary open angle glaucoma (POAG).</p><p><b>METHODS</b>Ninty-six cases (166 eyes) were randomly divided into an observation group (44 cases) and a control group (52 cases). The observation group was treated with pricking blood at Neiyingxiang (EX-HN 9), and the control group was treated with 0.5% Timolol Maleate drops in eyes. The changes of patients' intraocular pressure in both groups were observed before and after treatment.</p><p><b>RESULTS</b>The intraocular pressure were (29.81 +/- 3.49) mmHg and (23.18 +/- 3.61) mmHg before and after treatment in the observation group, and (29.94 +/- 3.64) mmHg and (23.88 +/- 3.96) mmHg in the control group respectively, there were significant differences after treatment in the two groups (both P<0.01), while there was no significant difference in decreasing the intraocular pressure between the two groups (P>0.05).</p><p><b>CONCLUSION</b>Pricking blood at Neiyingxiang (EX-HN 9) can effectively decrease the intraocular pressure and the effect is equivalent to Timolol Maleate drops.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Acupuncture Points , Bloodletting , Glaucoma, Open-Angle , Therapeutics , Intraocular PressureABSTRACT
<p><b>OBJECTIVE</b>To evaluate the risk factors of post-renal biopsy bleeding (PBB).</p><p><b>METHODS</b>We retrospectively analyzed the clinical data of 1 262 patients who received ultrasound-guided biopsy of native kidney at our hospital from January 2005 to December 2006.</p><p><b>RESULTS</b>The overall incidence of PBB was 30.3% (383/1,262), among which the incidence of hematoma was 29.4% (371/1,262) (the percentages of less and more than 5cm were 73.9% and 26.1%, respectively) while that of gross hematuria was only 1.3% (17/1,262). The incidences of minor, intermediate, and major bleeding complications were 21.4% (270/1,262), 8.4% (106/1,262), and 0.6% (7/1,262), respectively. In seven patients with major bleeding complications, six had renal disease secondary to rheumatic disease (lupus nephritis, n = 5; scleroderma crisis, n = 1), while the other one had IgA nephropathy (Lee's classification V). The risk of PBB was relatively higher in women and younger patients.</p><p><b>CONCLUSIONS</b>Patients with chronic connective tissue diseases are vulnerable to severe PBB complications. A close monitoring of these patients is necessary.</p>