Objective To investigate the impact of iron overload on apoptosis in primary mouse liver cells via a synchronous separation technology.Methods Hepatocyte(HC),liver sinusoidal endothelial cell(LSEC),and Kupffer cell(KC)were isolated and purified with collagenase,percoll density gradient centrifugation,and CD146 magnetic beads.Cell types were identified using flow cytometry and immunofluorescence staining.Cells of different types were cultured in vitro,and an iron overload model was established by treating the mice with 0,25,50 and 100 μmol/L ferric ammonium citrate(FAC)for 24 h.The iron content was quantified using Prussian blue staining,while cell viability and mitochondrial membrane potential were assessed by flow cytometry.Results The synchronous separation technology of primary liver cells exhibited stable efficiency.The yield of HC was(4.0±0.5)×107 cells per mouse,exhibiting an effective survival rate of(76.33±0.67)％.The yield of LSEC was(5.0±1.0)×106 cells per mouse,with a survival rate of(93.63±0.25)％and a purity level of(93.40±0.46)％.The yield of KC was(1.5±0.5)×106 cells per mouse while a high survival rate of(98.33±0.12)％and a purity level of(88.30±2.02)％were maintained.The obtained cells were large in number,with good vitality and high purity,which could meet the requirements of subsequent experiments.Treatment with FAC significantly elevated iron contents in different types of cells when compared with the control group.Upon stimulation of FAC,the survival rate of HC decreased from(73.97±5.54)％to(54.10±1.68)％,the mean fluorescence intensity of JC-1 aggregates decreased from 326.33±30.37 to 155.00±6.56,JC-1 monomer increased from 1700.00±1 44.04 to 3713.33±81.82.The survival rate of LSEC decreased from(90.60±1.74)％to(78.03±2.15)％,the mean fluorescence intensity of JC-1 aggregates decreased from 502.33±5.51 to 372.33±4.04,and JC-1 monomer increased from 750.00±67.51 to 1340.00±36.39.The survival rate of KC decreased from(94.23±1.44)％to(88.37±1.56)％,the mean fluorescence intensity of JC-1 aggregates decreased from 652.67±25.66 to 478.00±12.49,and JC-1 monomer increased from 1984.33±80.65 to 3062.33±245.20.Conclusion A robust and reliable simultaneous isolation technique of primary mouse HC,LSEC,and KC has been established.Moreover,our finding demonstrates that iron overload significantly enhances apoptosis levels in HC,LSEC and KC.

Parkinson's disease(PD)and multiple system atrophy(MSA)are two common Parkinsonian syndromes with overlapping clinical manifestations, and clinical differential diagnosis is difficult.Lower urinary tract symptoms are one of the common non-motor symptoms of the two diseases.The incidence of lower urinary tract symptoms in MSA is higher, the onset is earlier, and the micturition period is more prominent.The urinary dysfunction in patients with PD is mainly caused by the central mechanism, leading to overactive bladder.MSA has more extensive lesions with both central and peripheral involvement, leading to overactive bladder and severe voiding dysfunction.Urodynamics can be used to evaluate bladder and urethral function.MSA has more prominent weak detrusor activity, residual urine volume, and early changes of urethral sphincter.The treatment of lower urinary tract symptoms in patients with PD is mainly based on anticholinergic drugs to improve overactive bladder, while in MSA patients with increased residual urine volume, intermittent catheterization is the main method to improve lower urinary tract symptoms.This article reviewed the epidemiology, pathological mechanism, urodynamics and treatment of lower urinary tract symptoms of the two diseases, so as to assist in their differential diagnosis and treatment.

Objective:To investigate the incidence of various non-motor symptoms (NMS) in early stage of Parkinson′s disease (PD) patients and the differences between the body-first and brain-first subtypes.Methods:A total of 121 patients with PD (Hoehn-Yahr stage 1-2) were recruited from PD Clinic, Department of Neurology, Beijing Hospital from January 2012 to January 2015. The general information and clinical features of the patients were collected. The minimal diagnostic criteria of parasomnias described in the International Classification of Sleep Disorders-Revised were used to diagnose rapid eye movement sleep behavior disorder (RBD).According to the sequence of RBD and motor symptoms, the patients were divided into 2 groups: body-first subtype and brain-first subtype. NMS was evaluated by the Non-Motor Symptom Questionnaire (NMSQuest). The clinical features and the incidence of various NMS were compared between the 2 groups. The Unified Parkinson′s Disease Rating Scale (UPDRS) was used to evaluate the severity of the disease, and its third part (UPDRS-Ⅲ) was used to evaluate the motor function of the patients. Hamilton Rating Scale for Depression (HAMD) and Hamilton Rating Scale for Anxiety (HAMA) were used to evaluate the depression and anxiety status of the patients. The sleep status of patients was assessed by Parkinson′s Disease Sleep Scale (PDSS). The quality of life of the patients was assessed by 39-item Parkinson′s Disease Questionnaire (PDQ-39).Results:Of all the patients, 49.59% (60/121) had the body-first subtype and 50.41% (61/121) had the brain-first subtype of PD. There was no significant difference in UPDRS-Ⅲ score between the 2 groups. The average number of NMS in all PD patients was 10.97±4.88. Body-first subtype patients had higher NMS incidence than brain-first subtype in difficulty in swallowing [46.7% (28/60) vs 23.0% (14/61), χ 2=7.507, P=0.006], nausea and vomiting [16.7% (10/60) vs 3.3% (2/61), χ 2=6.069, P=0.014], constipation [85.0% (51/60) vs 55.7% (34/61), χ 2=12.393, P<0.001], fecal incontinence [8.3% (5/60) vs 0 (0/61), χ 2=5.302, P=0.021], difficulty in remembering recent events [58.3% (35/60) vs 32.8% (20/61), χ 2=7.962, P=0.005], loss of interest [43.3% (26/60) vs 24.6% (15/61), χ 2=4.743, P=0.029], inattention [45.0% (27/60) vs 19.7% (12/61), χ 2=8.884, P=0.003], depression [55.0% (33/60) vs 34.4% (21/61), χ 2=5.181, P=0.023], intense vivid dreams [73.3% (44/60) vs 39.3% (24/61), χ 2=14.196, P<0.001] and restless legs [53.3% (32/60) vs 27.9% (17/61), χ 2=8.140, P=0.004]. The differences were significant. Body-first subtype and NMSQuest ( r=-0.489, P<0.001), UPDRS ( r=-0.189, P=0.038), HAMD ( r=-0.231, P=0.011), HAMA ( r=-0.298, P=0.001) and PDQ-39 scores ( r=-0.276, P=0.002) were negatively correlated. Body-first subtype and PDSS score was positively correlated. NMSQuest (Δ R2=0.265, P<0.001) was the main determinant of PDQ-39 score. Conclusions:PD patients are accompanied by various NMS, which is a major factor affecting the quality of life. Compared with brain-first subtype, body-first subtype might have more NMS burden and higher incidence rate in most NMS in early PD patients.

Objective:To study the relationship between endoplasmic reticulum stress (ERS) and apoptotic protein and myocardial pathological changes in rats after endostar combined with low-dose X-ray irradiation.Methods:Forty SD rats were evenly divided into four groups: control group (intraperitoneal injection of equal volume physiological saline, once per day, 14 d), endostar group (intraperitoneal injection of endostar 6 mg/kg, once per day, 14 d), irradiation group (15 Gy divided into 3 times X-ray irradiation) and combination group (intraperitoneal injection of endostar after irradiation at the same dose and time as the endostar group). At 1 and 6 months after treatment, myocardial tissues of rats were prepared for HE staining and Masson staining to observe the myocardial histological changes. TUNEL assay was used to detect myocardial cell apoptosis, and ImageJ software was utilized to calculate myocardial collagen volume fraction (CVF). The expression levels of ERS and apoptotic protein glucose-regulated protein 78 (GRP78), protein kinase-like endoplasmic reticulum kinases (PERK), CCAAT/enhancer binding protein homologous protein (CHOP) and cysteine-containing aspartate-specific protease-12 (Caspase-12) were detected by Western blot. One-way ANOVA was conducted using GraphPad Prism 8.0.1 software, and comparison between two groups was conducted using t-test. Results:At 6 months after treatment, the myocardial interstitium in the irradiation and combination groups was widened, showing strip-like or reticular fibrosis changes, and the myocardial interstitium had diffuse collagen fiber deposition. Compared with the control group, CVF was increased significantly (both P<0.01). At 1 and 6 months after treatment, the apoptotic index of myocardial cells in the combination group was significantly higher than that in the control group ( P<0.05, <0.001). At 1 and 6 months after treatment, the expression levels of GRP78 protein in the irradiation and combination groups were increased (all P<0.01), and the expression levels of PERK and CHOP proteins in the combination group were increased compared to those in the control group (both P<0.05). At 6 months after treatment, the expression levels of PERK and CHOP proteins in the irradiation group were increased compared to those in the control group (both P<0.05). Compared with the control group, Caspase-12 expression levels at 1 and 6 months after treatment were increased in the endostar, irradiation and combination groups (all P<0.05). Conclusions:The expression levels of ERS and apoptotic proteins are related to cardiac injury caused by irradiation in rats. After low-dose X-ray combined with endostar treatment, ERS is aggravated and myocardial apoptosis is increased.

Objective:To investigate the implication of micro RNA-21(miR-21) in Endostar combined with X-ray irradiation of cardiac fibroblasts (CF).Methods:Rat CFs were used in this experiment and been divided into the blank control group, 10 Gy X-ray irradiation group, Endostar group, 10 Gy X-ray+ Endostar group, 10 Gy X-ray+ Endostar+ NC mimic group (negative control 1), 10 Gy X-ray+ Endostar+ miR-21 mimic group, 10 Gy X-ray+ Endostar+ NC inhibitor group (negative control 2) and 10 Gy X-ray+ Endostar+ miR-21 inhibitor group. The proliferation of CF was determined by Methyl thiazolyl tetrazolium (MTT) assay. The expression level of Collagen Ⅰ protein was analyzed by Western blot. The expression levels of Collagen Ⅰ and miR-21 mRNA were assayed by real-time quantitative polymerase chain reaction (q-PCR).Results:In the 10 Gy X-ray+ Endostar+ miR-21 mimic group, the CF proliferation, Collagen Ⅰ and miR-21 mRNA were increased significantly compared with those in the blank control group, 10 Gy X-ray+ Endostar group, and negative control group 1 (all P<0.05). In the 10 Gy X-ray+ Endostar+ miR-21 inhibitor group, the CF proliferation and expression levels of Collagen Ⅰ mRNA were decreased significantly compared with those in the blank control group, 10 Gy X-ray+ Endostar group and negative control group 2(all P<0.05). Conclusions:The CF proliferation and Collagen Ⅰ expression are increased when the expression level of miR-21 gene is simulated. When inhibiting the expression of miR-21 gene, the CF proliferation and Collagen Ⅰ expression are reduced.

Objective:To explore the establishment of radiation-induced heart damage (RIDH) SD rat models caused by irradiation of 15Gy/3f and the changes in early detection indicators, and evaluate the effect of irradiation combined with recombinant human endostatin (Endostar).Methods:75 adult male SD rats were randomly divided into the blank control group (C group), Endostar group (E group), 25Gy irradiation group (MHD 25 group), 15Gy irradiation group (MHD 15 group) and 15Gy irradiation combined with Endostar group (MHD 15+ E group), respectively. Blood sample was taken to measure the CK, CK-MB, LDH and CRP at 24h, 48h and 15d after corresponding interventions. After cardiac echocardiography at 1, 3 and 6 months, 5 rats in each group were randomly sacrificed and myocardial tissues were collected for HE and Masson staining. Two-way ANOVA was employed for statistical analysis. Results:Compared with group C, myocardial fibrosis were observed in the MHD 15 group at 6 months ( P<0.05), which occurred later than that in the MHD 25 group. Ejection fraction (EF) and fractional shortening (FS) were significantly decreased after 3 months in each irradiation group (all P<0.05), whereas the degree of decrease was similar among all groups (all P>0.05). The expression levels of myocardial enzymes and inflammatory cytokines did not significantly differ among different groups (all P>0.05). Conclusions:In the early stage, exposure to 15Gy/3f irradiation can cause cardiac function damage in SD rat hearts, such as the reduction of EF and FS, and even lead to myocardial fibrosis in the late stage, which is delayed and less severe than high-dose irradiation. Irradiation combined with Endostar has no significant effect on radiation myocardial injury in rats.

Objective:To investigate the clinical features and associated chronic pain in corticobasal syndrome (CBS).Methods:Clinical data of 8 patients diagnosed as probable CBS or possible CBS admitted to Beijing Hospital during January 2010 to June 2020 were retrospectively analyzed. The clinical information included sex, age, course of disease, chief complaint, neurological examination, blood biochemistry, tumor marker, infection and other laboratory tests; the neuropsychological evaluation included Mini-Mental State Examination (MMSE) scale and Hamilton Depression Scale (HAMD); the imaging studies included cranial magnetic resonance imaging (MRI) and/or 18F-Fluorodeoxyglucose positron emission tomography ( 18F-FDG PET). Results:The main clinical manifestations were asymmetrical movement disorders, including rigidity, tremor, myoclonus and abnormalities in posture and gait. Patients showed poor response to levodopa treatment. Among 8 patients, 7 had apraxia, 5 patients had alien hand, and 5 patients had various degrees of cognitive dysfunction. The cranial MRI demonstrated mild cerebral atrophy which was slightly more severe in the contralateral side of the initially affected limb in 7 of the 8 patients. The 18F-FDG PET scan revealed asymmetric decreased metabolism in the frontal, parietal, temporal, and occipital lobe, as well as in basal ganglia, which was more severe in the contralateral side of the initially affected limb in 5 of the 8 patients. Six of the 8 patients were associated with pain, including dystonic pain in 3 patients, neuropathic pain in 1 patient, musculoskeletal pain in 1 patient, and unexplained pain in 1 patient. Pain was the onset symptom in 1 patient and pain was relieved by taking levodopa in another patient. Conclusions:CBS is characterized by asymmetric dyskinesia and cognitive impairment, and often associated with apraxia, cortical sensory deficits, and alien limb. The MRI and PET are helpful for CBS diagnosis. Pain may be one of the common non-motor symptoms in CBS.

Objective: To survey the prevalence and distribution of sleep disorders in patients with Parkinson disease (PD) and to analyze the influencing factors. Methods: The prevalence and distribution of sleep disorders were surveyed with Parkinson Disease Sleep Scale (PDSS) among 206 PD patients. The association of sleep disorders with age, course of disease, cognitive function, motor function, depression, and the equivalent dose of levodopa (LED) was analyzed. Results: The overall PDSS score in 206 patients was (116.9±21.4). The three most frequent items of sleep disorders were the overall sleep quality(181/206, 87.9%), difficulty in maintaining sleep(160/206, 77.7%)and nocturnal enuresis(151/206, 73.3%); the three least frequent items were early awaking(87/206, 42.2%), urinary incontinence(56/206, 27.2%)and hallucination(44/206, 21.4%). The three items with the lowest average scores were nocturnal enuresis(6.9±3.1), difficulty in maintaining of sleep(7.1±2.7)and overall sleep quality(7.1±2.0); three items with the highest average scores were audiovisual illusion(9.3±1.8), incontinence caused by motion disability(9.0±2.1) and early awaking with upper and lower limb pain(8.7±2.1). PD patients were divided into group 1 [Hoehn-Yahr(H&Y) stage 1.0-1.5], group 2 (H&Y stage 2.0-2.5) and group 3 (H&Y stage 3.0-4.0). One-way analysis of variance or non-parametric test showed that there were significant differences in the course of disease(F=21.91, P=0.00), total score and the subscale scores of Unified Parkinson Disease Rating Scale(UPDRS, UPDRS Ⅰ-Ⅳ) (F=90.67, χ²=12.86, F=31.20, F=60.17, χ²=24.01, all P<0.01), the Hamilton Rating Scale for Depression(HAMD) scores (χ²=11.06, P=0.00), LED (F=14.93, P=0.00) and Minimum Mental State Examination(MMSE) scores (χ²=9.81, P=0.01) among three groups. There was no significant difference in age and PDSS scores among three groups. The results showed that there were significant differences in terms of restless state and nocturnal dysphoria (F=5.12, P=0.01; F=3.27, P=0.04) between group 1 and group 3. The linear regression analysis showed that the HAMD and the LED scores had the greatest influence on PDSS score (R²=0.142, 0.196). Conclusion PD patients have a variety of sleep symptoms. The treatment of large doses of dopamine and depression contribute to the occurrence of PD sleep disorders.

Objective To explore the clinical effect of continuous venous-venous hemofiltration (CVVH) combined with hemoperfusion (HP) in treatment of patients with hypertriglyceridemia pancreatitis (HTGP). Methods The clinical data of 33 patients with moderate and severe HTGP who were treated by CVVH combined with HP were retrospectively analyzed from March 2012 to March 2017 in Wuhan general hospital of the people's liberation army. The differences of vital signs and the serum levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL), amylase (AMS), interleukin-6 (IL-6), blood calcium (Ca2+) and white blood cell count (WBC), haemoglobin (Hb), platelet count (PLT) before and 24 hours,72 hours and 1 week after therapy were compared, the changes of recovery time to target serum TG level, frequency of blood purification therapy, time for disease situation becoming stable, days staying in hospital and mortality were observed. Results The levels of LDL were not high in patients with HTGP, the levels of TG and TC were decreased significantly after using CVVH plus HP, and after treatment for 24 hours statistical differences appeared compared with those before treatment [TG (mmol/L):7.14±1.04 vs. 11.90±2.03, TC (mmol/L): 7.47±1.04 vs. 10.20±1.26], the decline persisting to 1 week after treatment;the drop rates of TG and TC were the largest after the first combined treatment, and the TG drop rate was more obvious than that of TC [(51.92±14.18)% vs. (30.09±10.01)%, P < 0.05], an average of (2.58±1.45) days and (2.38±0.98) times of combined blood purification could restore the TG to its safe level (TG < 5.65 mmol/L), the time of disease situation tending to be stable was (7.46±3.05) days and the time of staying in hospital was (20.00±2.12) days. Systemic inflammatory response syndrome (SIRS) related vital signs and inflammatory response indicators were also improved obviously after the combined therapy (all P < 0.05), after treatment for 72 hours, various vital signs and Ca2+reached to their normal reference ranges, after treatment for 24 hours IL-6 began to decline significantly compared with that before treatment (ng/L: 120.85±16.45 vs. 151.05±18.19), and AMS and WBC returned to their normal reference ranges after treatment for 1 week. Conclusion CVVH combined with HP can quickly and effectively eliminate TG in blood in patients with HTGP and in the mean time it may ameliorate and block the early progression of SIRS, resulting in good therapeutic effect on alleviating the disease development and improving its prognosis.

Objective To study the effects of hemodialysis (HD) combined with hemoperfusion (HP) on sleep quality in maintenance hemodialysis (MHD) patients. Methods Sixty MHD patients admitted to Department of Blood Purification of Wuhan General Hospital of PLA from January to December 2016, 30 cases were treated with HD, and the other 30 cases were treated by HD+HP, the course of treatment was 12 weeks in both groups. The changes of serum β2-microglobulin (β2-MG) and parathyroid hormone (iPTH) were observed before treatment and 12 weeks after treatment; the sleep quality of all patients in the two groups were evaluated by Pittsburgh Sleep Quality Index (PSQI) Scale, and the correlations between the sleep quality of MHD patients andβ2-MG level, iPTH level were analyzed by Pearson linear correlation analysis. Results All the 60 patients completed the treatment. The serum β2-MG, iPTH levels and PSQI score after treatment were decreased obviously in HD+HP group compared with those before treatment, and the degrees of decrease in HD+HP group were more significant than those in the HD group [β2-MG (mg/L): 12.34±2.12 vs. 20.27±3.15, iPTH (ng/L): 224.54±100.28 vs. 398.42±155.37, PSQI score:8.56±0.86 vs. 12.45±0.88, all P < 0.05]. Pearson linear correlation analysis showed that the PSQI score was significantly positively correlated with serum β2-MG, iPTH level (r respectively was 0.416 and 0.462, both P < 0.01). Conclusion HD+HP therapy can significantly improve the sleep quality of MHD patients, and the mechanism may be related to the elimination of serum iPTH and β2-MG from the body of MHD patients.