Objective Electric stimulation functional observation statistics (FES) were collected for elderly patients with cerebral apoplexy in search of any curative effect. Methods A total of 40 patients with cerebral apoplexy were divided randomly into a FES group of 20 cases and a control group of 20 cases. Both groups received the same routine rehabilitation training and basic drugs. The FES group was also treated using FES therapeutic apparatus for 30 min daily over 3 weeks (15 sessions). The control group received no electrical stimulation. Upper limb motor function, lower limb motor function, balanceand Barthel's index were evaluated. Results In the FES group, upper limb motor function was significantly improved after treatment, and significantly better compared with the control group. Lower limb motor function showed the same significant differences. Balance was also significantly better after treatment and significantly better than in the control group. Similar significant improvements were seen in terms of Barthel's index. Conclusions FES can improve the functional capacity of elderly patients with cerebral apoplexy hemiplegia and improve their ability in daily activities.

Objective:To investigate the effects of pulmonary rehabilitation on anxiety/depression and subjective/objective sleep quality of elderly patients with stable chronic obstructive pulmonary disease(COPD).Methods:From February 2018 to February 2019, 120 elderly patients with stable COPD were selected and randomly divided into the experimental group (pulmonary rehabilitation exercise combined with conventional COPD treatment) and the control group (simple COPD conventional treatment). Sixty cases in each group were intervened for 8 weeks. Before and after treatment, Hamilton anxiety scale (HAMA) was used to evaluate anxiety, Hamilton depression scale(HAMD)was used to evaluate depression, Pittsburgh sleep quality index(PSQI)and sleep log were used to evaluate subjective sleep quality, and objective sleep quality was monitored by multi-channel sleep monitor.SPSS 21.0 software was used to analyze and process the data. Chi square test, independent sample t test and paired t test were used for statistical analysis. Results:After 8 weeks of intervention, the HAMA and HAMD scores of the experimental group were lower than those of the control group (HAMA: (7.57±3.19) vs (10.15±4.89), t=-3.428, P=0.001; HAMD: (8.22±4.73) vs (10.60±6.49), t=-2.300, P=0.023). COPD patients with anxiety decreased (χ 2=7.566, P=0.006). After treatment, the subjective sleep latency of the experimental group was shorter than that of the control group ((42.00±9.88)min vs (47.25±10.27)min, t=-2.854, P=0.005). The subjective sleep efficiency was higher than that of the control group ((76.00±4.50)% vs (74.00±5.20)%, t=2.272, P=0.025), and the objective sleep latency was shorter than that of the control group ((28.02±5.59)min vs (32.95±6.21)min, t=-4.575, P<0.05). Conclusion:Pulmonary rehabilitation exercise can improve the anxiety and depression of elderly patients with stable COPD, and improve the subjective and objective sleep quality.

Objective To search MRI features of patients with HIV-negative cryptococcus meningoencephalitis (CM)and evaluate the value in judging the prognosis of CM.Methods The findings of cranial MRI and prognosis were retrospectively analyzed in 19 cases with India-ink capsule staining-proved HIV-negative CM in our department.The prognosis was evaluated by Glasgow Outcome Scale:score 1=death,score 2=persistent vegetative state,score 3=severe disability,score 4=moderate disability,score 5=good recovery. The patients with score 1-3 were classified as the group of poor prognosis,while those with score 4,5 as good prognosis.Results Abnormal manifestations on cranial MRI were observed in 84.2%(1 6/1 9)cases,meninges enhanced in 84.2% (1 6/1 9),brain edema in 31.6%(6/1 9),brain parenchyma lesion in 47.4% (9/1 9 ),vasculitis in 5.2% (1/1 9 ).The mean duration of follow-up was 3.5 years,The prognosis of 5/9(44.4%)cases with brain parenchyma lesion confirmed by cranial MR was poor.Conclusion The MRI manifestation of HIV-negative CM is diverse,and brain parenchyma lesion confirmed by cranial MRI may associated with poor prognosis.

The purpose of this study was to study the effect of three different transfection reagents (Lipofectamine™ LTX & PLUS™, Lipofectamine 2000 and Nano-PAMAM-D) and three different testicular injection methods (rete testicular injection, seminiferous tubules injection and testicular interstitial injection) on the efficiency of production transgenic mice. After the mixtures of plasmid DNA (pEFP-C1) and transfection reagent were injected with different testicular injection methods, the sperm density, vitality, positive sperm rates and PCR positive transgenic mice rate were examined 30 days after injection. The results showed that the damage degree from slight to serious of three transfection reagents was Lipofectamine™ LTX & PLUS™, Lipofectamine 2000, and PAMAM-D. The sperm positive rates with green fluorescence of these three groups were 35.65%±0.69%, 12.86%±0.35% and 10.04%±0.20%, respectively. The PCR positive rates of transgenic newborn mice were 29.17%, 13.70% and 5.88%, respectively. Among the groups of different testicular injection methods, the damage degree from slight to serious was rete testicular injection, seminiferous tubules injection, and testicular interstitial injection, whereas the sperm positive rates with green fluorescence were 35.13%, 15.13%, and 0%, respectively. The PCR positive rates of transgenic newborn mice among different testicular injection groups were 33.3%, 12.5%, and 0.0%. The combination of rete testicular injection and Lipofectamine™ LTX & PLUS™ had the lowest toxicity and highest transgenic efficiency in the production of transgenic mice.
Animals ; Humans ; Indicators and Reagents ; chemistry ; Injections ; methods ; Lipids ; chemistry ; Male ; Mice ; Mice, Transgenic ; Plasmids ; Polymerase Chain Reaction ; Seminiferous Tubules ; Spermatozoa ; Testis ; Transfection

Objective To explore the relationship between pyroptosis and treatment in non-small cell lung cancer patients treated with tyrosine kinase inhibitors targeted therapy. Methods Stable transfection strains with common EGFR mutations found in clinical practice were constructed through lentiviral transfection. LDH and Western blot experiments were conducted to determine the degree and mechanism of pyroptosis after osimertinib treatment. Animal experiments verified the effect of pyroptosis on treatment efficacy. ELISA was used to explore the potential connection between pyroptosis and tumor immunotherapy. Results After osimertinib treatment on stable lines, the EGFR-L858R mutation had obvious pyroptosis at the morphology and protein levels. Western blot experiment confirmed that pyroptosis was mediated by GSDME (P < 0.0001). Experiments through the overexpression of GSDME and corresponding animal studies discovered that the degree of pyroptosis affected the treatment outcome. Blood analysis revealed that the level of IL-1β secreted by EGFR-L858R and EGFR-L858R-GSDME-OE mice after treatment was higher than that of the control group (P < 0.0001), and it may regulate tumor immunity to a certain extent. Conclusion Osimertinib can induce pyroptosis in EGFR-L858R mutant strains mediated by GSDME, and the level of pyroptosis in cell lines is positively correlated with therapeutic effect to a certain extent.

Objective To evaluate whether the reverse phase protein array (RPPA) method can be used for detecting TORCH infection in pregnant women .Methods The RPPA method was established for detecting TORCH infection .The positive coinci‐dence rates of TORCH infection detected by the RPPA method and ELISA method in 2000 fresh serum samples from pregnant women were compared for evaluating the feasibility of RPPA in TORCH detection .Results The positive coincidence rates of estab‐lished RPPA and ELISA for detecting TORCH infection was 100 .0% ,91 .1% ,97 .2% ,91 .3% and 93 .0% respectively ,indicating that the detection results of various indexes by RPPA and ELISA had better consistency (P>0 .05) ,but the positive detection rates of RPPA for Rubellavirus ,CMV and HSV‐1 ,2 were higher than those of correspondent ELISA method .Conclusion RPPA method for detecting TORCH infection has the advantages of simpleness ,rapidness ,high sensitivity and strong specificity ,is an effective method of auxiliary diagnosis for bearing and rearing better children in clinical ,and is worthy of being promoted and used in the fu‐ture .

This study aims to construct the recombinant lentivirus vector containing specific small interfering RNA (siRNA) targeting rat CREB binding protein(CBP)gene and to identify its function of inhibiting the expressions of acetylated histone in primarily cultured hippocampal neurons. Firstly, we constructed four kinds of recombinant lentivirus siCBP. And then we used them to infect the primarily cultured hippocampal neurons, and performed real-time PCR, western blot respectively to detect the expressions of CBP. Afterwards, the most effective lentivirus siCBP was used to infect the primarily cultured hippocampal neurons, and then the HAT activity and protein expressions of acetylated histone Ac-H3, Ac-H4 of the neurons were examined. By using PCR, endonuclease cutting and gene sequencing, we confirmed that the target genes were correctly cloned in lentivirus vector. Besides, CBP mRNA and protein expressions in neurons were found to be with varying degrees of decreases after infections of the four kinds of lentivirus siCBP. Furthermore, the representative and most effective lentivirus GR806 could effectively inhibit the HAT activity and the protein expressions of Ac-H3, Ac-H4 in neurons. It provides the experimental basis for the subsequent application of siCBP to clarify the effects and corresponding molecular mechanism of the CBP-dependent histone acetylation on learning and memory function in hippocampus.
Acetylation ; Animals ; CREB-Binding Protein ; metabolism ; Genetic Vectors ; Hippocampus ; cytology ; Histones ; metabolism ; Lentivirus ; Memory ; Neurons ; metabolism ; Primary Cell Culture ; RNA, Messenger ; RNA, Small Interfering ; Rats ; Real-Time Polymerase Chain Reaction

OBJECTIVETo investigate the expression of acetyl coenzyme A synthetase short-chain family member 2 (ACSS2) in patients with colorectal cancer (CRC) and its biological role.

METHODSA total of 74 CRC tissue samples and 40 normal colorectal tissues were tested by immunohistochemical staining to detect the expression of ACSS2 (cell staining intensity score: 0 points: without staining, 1 points: weak staining, 2 points: intensity staining, 3 point: strong staining; the percentage of positive cells in tumor or negative score: 0 points: negative, 1 point: <25% positive cells, 2 points: 25%-50% positive cells, 3 points: 50%-75% positive cells, 4 points: >75% positive cells. The product of above two scores was the final score.). Association of ACSS2 expression with clinicopathological characteristics was analyzed. Small interfering RNA (siRNA, including A and B group) was used to knock down the expression of ACSS2 in colorectal cell lines (Lovo, HCT116) and their proliferation, migration and epithelial-mesenchymal transition (E-cadherin and Snail as markers) after knocking down were observed.

RESULTSThe expression of ACSS2 was significant higher in CRC tissue than that in normal colorectal tissue (tumor average score 6.284, normal tissue average score 3.625, P<0.01) and the percentage of positive cell was higher than that in normal tissue (tumor 69.9%, normal tissue 45.1%, P=0.000). The use of ACSS2 siRNA successfully knocked down the expression of ACSS2 in Lovo and HCT116 cells. A mild suppression of cell proliferation was observed 5 days after planked (A450 value: Lovo-NC 1.758±0.041, Lovo-ACSS2-siA 1.485±10.026, Lovo-ACSS2-siB 1.371±0.049; HCT116-NC 2.609±0.038, HCT116-ACSS2-siA 2.260±0.042, HCT116-ACSS2-siB 2.295±0.029). While a remarkable ability decline of cell migration was found (Lovo-NC 225±5/field, Lovo-ACSS2-siA 40±5/field, Lovo-ACSS2-siB 79±3/field; HCT116-NC 198±7/field, HCT116-ACSS2-siA 96±7/field, HCT116-ACSS2-siB 77±9/field, P<0.05). Real-time quantitative PCR detection showed that in Lovo cells, expression of E-cadherin up-regulated and expression of Snail down-regulated, while in HCT116 cells, E-cadherin up-regulated slightly [E-cadherin: Lovo NC 1.000±0.211, Lovo-siA 3.403±0.207, Lovo-siB 2.658±0.420 (P<0.05); HCT116 NC 1.000±0.121, HCT116-siA 1.349±0.197, HCT116-siB 1.528±0.147(P>0.05); Snail: Lovo NC 1.000±0.085, Lovo-siA 0.468±0.030, Lovo-siB 0.499±0.088 (P<0.05); HCT116 NC 1.000±0.118, HCT116-siA 0.265±0.020, HCT116-siB 0.194±0.017 (P<0.05)].

CONCLUSIONCRC tissues have high expression of ACSS2, which may be associated with cell migration and epithelial-mesenchymal transition.

Liver failure is a kind of acute and severe liver disease.Bone marrow-derived mesenchymal stem cells(BM-MSCs)have the function of differentiating into hepatocytes.Promotion of the regeneration of hepatocytes re-generation,inhibition of apoptosis,necrosis and inflammation of hepatocytes,may facilitate repairing damaged liver tissue and improving liver function.BM-MSCs have become a new choice with great application potential in the treatment of liver failure.

Arsenic compounds are widely used for the therapeutic intervention of multiple diseases.Ancient pharmacologists discovered the medicinal utility of these highly toxic substances,and modern phar-macologists have further recognized the specific active ingredients in human diseases.In particular,Arsenic trioxide(ATO),as a main component,has therapeutic effects on various tumors(including leukemia,hepatocellular carcinoma,lung cancer,etc.).However,its toxicity limits its efficacy,and con-trolling the toxicity has been an important issue.Interestingly,recent evidence has pointed out the pivotal roles of arsenic compounds in phase separation and membraneless organelles formation,which may determine their toxicity and therapeutic efficacy.Here,we summarize the arsenic compounds-regulating phase separation and membraneless organelles formation.We further hypothesize their potential involvement in the therapy and toxicity of arsenic compounds,highlighting potential mecha-nisms underlying the clinical application of arsenic compounds.