Overexpression of circular RNA HIPK3 prevents activation of rat microglia / 安徽医科大学学报

Objective @#To investigate the relationship between circular RNA homeodomain interacting protein kinase 3 (circHIPK3) and the activation of rat microglia (RM) cells.@*Methods @#In vitro , RM cells were cultured and randomized into normal and oxygen⁃glucose deprivation/reperfusion (OGD/R) groups , and the expression level of circHIPK3 in each group was detected by RT⁃qPCR. The circHIPK3 lentiviral vector with puromycin resistance was constructed , and the overexpression (OE) group and negative control (NC) group were set up. The optimal multiplicity of infection (MOI) for RM cells was determined based on fluorescence expression , and puromycin was used to screen RM cells stably expressing circHIPK3. The cells of OE and NC groups were treated with OGD/R , and the expression levels of ionized calcium binding adaptor molecule 1 (Iba ⁃1) and eukaryotic tumor necrosis factor receptor superfamily (CD40) were detected by Western blot. The circHIPK3 translational protein potential was analyzed by the circRNAdb database , while the potential binding microRNAs on circHIPK3 were predicted by circBank and Starbase databases.@*Results @#OGD/R down⁃regulated circHIPK3 in RM cells ( P < 0. 000 1) . The sequencing results were accurate and the lentiviral vector of circHIPK3 was constructed successfully. The optimal MOI of RM cells was 80 , puromycin at a concentration of 2 μg/ml was used to screen RM cell lines stably expressing circHIPK3. RT⁃qPCR results showed that the expression level of circHIPK3 was significantly higher in the OE group compared with the NC group (P < 0. 01) . Western blot results revealed that the expression levels of Iba and CD40 in the OE group were markedly lower than those in the NC group (P < 0. 05) . Protein translation analysis showed that circHIPK3 encoded a polypeptide of 404 amino acids with two internal ribosome entry sites (IRES) and an open reading frame (ORF) . Database analysis uncovered that circHIPK3 could target eight specific miRNAs , namely hsa⁃miR⁃3529⁃5p , hsa⁃miR⁃379⁃5p , hsa⁃miR⁃506⁃3p , hsa⁃miR⁃33 , hsa⁃miR⁃450b⁃5p , hsa⁃miR551b⁃3p , hsa⁃miR⁃193 , and hsa⁃miR⁃508 ⁃3p.@*Conclusion @#The overexpression of circHIPK3 effectively suppresses OGD/R⁃induced activation of RM cells. It has the potential to encode peptides and may act as a miRNA sponge. These findings provide a foundation for further study of circHIPK3 functions.

Effect of nasal swell body on nasal airflow and Artemisia pollen deposition / 临床耳鼻咽喉头颈外科杂志

Objective:The nasal swell body（NSB） consists of the nasal septal cartilage, nasal bone, and swollen soft tissue, all of which are visible during endoscopic and imaging examinations. Although the function of the NSB remains uncertain, there is evidence to suggest that it plays a vital role in regulating nasal airflow and filtering inhaled air. Based on anatomical and histological evidence, it is hypothesized that the NSB is indispensable in these processes. This study aims to investigate the impact of NSB on nasal aerodynamics and the deposition of allergen particles under physiological conditions. Methods:The three-dimensional （3D） nasal models were reconstructed from computed tomography （CT） scans of the paranasal sinus and nasal cavity in 30 healthy adult volunteers from Northwest China, providing basis for the construction of models without NSB following virtual NSB-removal surgery. To analyze the distribution of airflow in the nasal cavity, nasal resistance, heating and humidification efficiency, and pollen particle deposition rate at various anatomical sites, we employed the computed fluid dynamics（CFD） method for numerical simulation and quantitative analysis. In addition, we created fully transparent segmented nasal cavity models through 3D printing, which were used to conduct bionic experiments to measure nasal resistance and allergen particle deposition. Results:①The average width and length of the NSB in healthy adults in Northwest China were （12.85±1.74） mm and （28.30±1.92） mm, respectively. ②After NSB removal, there was no significant change in total nasal resistance, and cross-sectional airflow velocity remained essentially unaltered except for a decrease in topical airflow velocity in the NSB plane. ③There was no discernible difference in the nasal heating and humidification function following the removal of the NSB; ④After NSB removal, the deposition fraction（DF） of Artemisia pollen in the nasal septum decreased, and the DFs post-and pre-NSB removal were（22.79±6.61）% vs （30.70±12.27）%, respectively; the DF in the lower airway increased, and the DFs post-and pre-NSB removal were（24.12±6.59）% vs （17.00±5.57）%, respectively. Conclusion:This study is the first to explore the effects of NSB on nasal airflow, heating and humidification, and allergen particle deposition in a healthy population. After NSB removal from the healthy nasal cavities: ①nasal airflow distribution was mildly altered while nasal resistance showed no significantly changed; ②nasal heating and humidification were not significantly changed; ③the nasal septum's ability to filter out Artemisia pollen was diminished, which could lead to increased deposition of Artemisia pollen in the lower airway.

Application of innovative PBL in pathophysiology teaching / 中华医学教育探索杂志

Pathophysiology is a comprehensive subject, which is very important to cultivate the clinical comprehensive thinking of medical students. Pathophysiology involves a wide range of subjects and contents, and is one of the major and difficult courses in basic medicine. Based on many years of research and practice, we have developed main-line-problem-based learning (ML-PBL). ML-PBL is a diversified teaching mode, including the main line teaching method, clinical case discussion method, and the main line synopsis explaining method. The analysis of application results shows that ML-PBL teaching improves the student performance. Furthermore, ML-PBL is more conducive to cultivating the comprehensive clinical thinking ability, improving the independent learning ability, and fully mobilizing the learning initiative.

Application of Region 4 Stork system in the neonatal screening for very long chain acyl-CoA dehydrogenase deficiency by tandem mass spectrometry / 中华检验医学杂志

Objective:To investigate the application feasibility of Region 4 Stork (R4S) system, an international collaborative newborn screening data platform, combined with cut-off value analysis in the neonatal screening for very long chain acyl-CoA dehydrogenase deficiency (VLCADD) by tandem mass spectrometry (MS/MS).Methods:The retrospective study was performed in 2, 040 072 neonates screened by MS/MS in Neonatal Screening Center of Zhejiang Province, China from October 2013 to July 2018. Nine hundred and ten cases were determined and identified as suspected positive VLCADD neonates by traditional cut-off method of tandem mass spectrometry. The original data of these 910 screened neonates were further analyzed by R4S system. Based on clinical diagnosis and ACADL gene test results, the screening efficiency between two methods was statistically compared.Results:The data of 910 suspected VLCADD-positive cases interpreted by cut-off method were further analyzed by R4S system, and the positive interpretation was reduced to 238 cases (including 9 confirmed positive cases). A total of 16 different mutations were found in ACADL gene sequencing among the confirmed children. The screening false positive rate (FPR) declined from 0.44‰ (901/2 040 072) to 0.11‰ (229/2 040 072), the rate of positive predictive value (PPV) increased from 0.99% (9/910) to 3.78% (9/238), and the specificity increased from 99.96% (2 039 162/2 040 063) to 99.99% (2 039 834/2 040 063). There was a statistically significant difference between cut-off method alone and cut-off method combined R4S system analysis (χ2=393.5, P<0.05). Conclusions:The R4S system combined with cut-off method applied in VLCADD neonatal screening by MS/MS can effectively improve screening performance, reduce false positive rate, and has certain value in clinical application.

Screening and clinical analysis of isovaleric acidemia newborn in Zhejiang province / 浙江大学学报·医学版

OBJECTIVE@#To investigate the incidence,clinical,biochemical and genetic characteristics of isovaleric acidemia (IVA) in Zhejiang province.@*METHODS@#Between January 2009 and December 2019, a total of 3 510 004 newborns were screened for IVA using tandem mass spectrometry. Patients of IVA were confirmed by urine organic acid and @*RESULTS@#A total of 15 patients with IVA were diagnosed, with an incidence of 1/234 000. Three patients had acute neonatal IVA, and the rest were asymptomatic. The isovalerylcarnitine (C5) levels were increased in all patients. Twelve children underwent urinary organic acid analysis, of which 11 cases had elevated isovalerylglycine levels, 4 cases with 3-hydroxyisovalerate increased simultaneously. Eleven IVA patients underwent genetic testing, 9 patients were compound heterozygous variants in @*CONCLUSIONS@#The clinical manifestations of IVA are non-specific, and the gene spectrum is scattered. Newborn patients screened by tandem mass spectrometry can receive early diagnosis and treatment, so as to correct metabolic defects and pathophysiological changes.

Effects of delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots / 浙江大学学报·医学版

OBJECTIVE@#To explore effects of different delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots (DBS), so as to provide evidence for improving accurate and reliable detection by tandem mass spectrometry.@*METHODS@#A total of 1 254 616 newborn DBS samples in Newborn Screening Center of Zhejiang Province were delivered and stored at room temperature (group A, @*RESULTS@#The concentrations of amino acids and carnitines in the three groups were skewed, and the differences in amino acid and carnitine concentrations among groups were statistically significant (all @*CONCLUSIONS@#Cold-chain logistics system and storage in low temperature and low humidity can effectively reduce degradation of some amino acids and carnitines in DBS, improve the accuracy and reliability of detection, and thus ensures the quality of screening for neonatal metabolic diseases.

Sleep-Aids Derived from Natural Products

Although drugs such as barbiturates and benzodiazepines are often used for the treatment of insomnia, they are associated with various side effects such as habituations, tolerance and addiction. Alternatively, natural products with minimal unwanted effects have been preferred for the treatment of acute and/or mild insomnia, with additional benefits of overall health-promotion. Basic and clinical researches on the mechanisms of action of natural products have been carried out so far in insomnia treatments. Recent studies have been focusing on diverse chemical components available in natural products, with an interest of developing drugs that can improve sleep duration and quality. In the last 15 years, our co-workers have been actively looking for candidate substances from natural products that can relieve insomnia. This review is, therefore, intended to bring pharmacological data regarding to the effects of natural products on sleep duration and quality, mainly through the activation of GABAA receptors. It is imperative that phytochemicals will provide useful information during electroencephalography (EEG) analysis and serve as an alternative medications for insomnia patients who are reluctant to use conventional drugs.

Cocaine- and Amphetamine-Regulated Transcript (CART) Peptide Plays Critical Role in Psychostimulant-Induced Depression

Cocaine- and amphetamine-regulated transcript (CART) peptide is a widely distributed neurotransmitter expressed in the central nervous systems. Previously, several reports demonstrated that nucleus accumbal-injected CART peptide positively modulated behavioral sensitization induced by psychostimulants and regulated the mesocorticolimbic dopaminergic pathway. It is confirmed that CART peptide exerted inhibitory effect on psychostimulant-enhanced dopamine receptors signaling, Ca2+/calmodulin-dependent kinase signaling and crucial transcription factors expression. Besides modulation of dopamine receptors-related pathways, CART peptide also exhibited elaborated interactions with other neurotransmitter receptors, such as glutamate receptors and γ-aminobutyric acid receptors, which further account for attribution of CART peptide to inhibition of psychostimulant-potentiated locomotor activity. Recently, CART peptide has been shown to have anxiolytic functions on the aversive mood and uncontrolled drug-seeking behaviors following drug withdrawal. Moreover, microinjection of CART peptide has been shown to have an anti-depressant effect, which suggests its potential utility in the mood regulation and avoidance of depression-like behaviors. In this review, we discuss CART pathways in neural circuits and their interactions with neurotransmitters associated with psychostimulant-induced depression.

Preliminary application of Region 4 Stork project used in newborn screening by tandem mass spectrometry / 中华检验医学杂志

Objective To investigate the feasibility of Region 4 Stork(R4S)project used for newborn screening by tandem mass spectrometry in China.Methods This retrospective study was performed among 362 822 neonates screened by tandem mass spectrometry from May 2015 to April 2016 in Zhejiang newborn screening center.Infants were grouped by screening result category: 83 true positive cases,360 554 true negative cases and 2 185 false positive cases.Raw data was uploaded into R4S website to perform postanalytical interpretive tools, then results were analyzed with interpretation rules.The comparisons of normal population percentiles were done at five selected percentiles between Zhejiang newborn screening center and R4S project with min-max normalization.Results Compared with cutoff system by using R4S project with interpretation rules,the positive predictive value increased from 3.7%to 8.3%,the specificity increased from 99.40%to 99.75%, and the false positive rate declined from 0.6% to 0.2%. The two cases of true positive hyperprolinemia were reported negative, and one case of β-ketothiolase deficiency was misdiagnosis.Totally 311 638 cases in true negative group were resolved by postanalytical interpretive tools,and the remaining 48 916 cases were excluded with interpretation rules.False positive cases were reduced to 897 cases.Results of percentiles comparison showed that levels of some markers were significantly different between zhejiang newborn screening center and R 4S project.Conclusions R4S project effectively improved the newborn screening performance, whereas leaded to a small number of misdiagnosis and missed diagnosis.Besides,many true negative cases should be excluded with interpretation rules.Optimization should be achieved based on local normal population.(Chin J Lab Med,2018,41:300-304)

The correlation between MSCT enhanced findings and pathologic risk in the patients with primary small intestinal stromal tumor / 实用放射学杂志

Objective To analyze the multi-slice computed tomography (MSCT) enhanced findings in the patients with primary small intestinal stromal tumor(SIST),and to probe the relationship between the imaging findings and the pathologic risk in order to improve the diagnostic accuracy.Methods Thirty patients with primary SIST confirmed by surgical pathology were enrolled in this study.Characterization and compassion of the clinical manifestations and MSCT enhanced findings were carried out between the pathologic low-and high-risk groups.Furthermore,the relationship was analyzed between the enhanced findings and the pathologic risk.Results Among all 30 patients with primary SIST,the lesion was located at duodenum in 5 patients (16.7％),at jejunum in 16 (53.3％),and at ileum in 9 (30％).14 patients were classified in the low risk group with the lesion with the average length of (3.8±0.9) cm,and other 16 in the high-risk group with lesion with the average length of (7.0 ± 1.4) cm.There were no statistical differences between the low-and high-risk groups in CT value in plain and venous phase,and in added value in arterial,venous and delayed phases.However,the significantly differences were observed in CT value in arterial and delayed phases between two groups (P＜0.05).Conclusion MSCT may effectively evaluate the pathologic risk of primary SIST.There are significant differences of the enhanced findings between the low and high-risk groups,which can provide important apreoperative classification for the therapy.

Effects of physical and chemical factors in environment for dried blood spot preparation of neonatal screening assay / 临床检验杂志

Objective To investigate the effects of physical and chemical factors in the environment for dried blood sample (DBS) preparation of neonatal screening assay.Methods A total of 60 normal and 120 positive DBS were prepared under control and 10 different conditions.Another 30 normal and 80 positive DBS were prepared under control and 7 different concentration gradients of formaldehyde.The levels of phenylalanine (Phe),glucose-6-phosphate dehydrogenease (G6PD),thyroid stimulating hormone (TSH) and 17α-hydoxyprogesterone (17α-OHP) were tested by time-resolved fluorescence immunoassay or fluorescence assay.Statistical analysis was performed using SPSS 22.0 software.Results Compared with the control group,the results of Phe were not significantly different (P ＞ 0.05) when the samples were dried under the formaldehyde sensitive threshold (4.62 to 6.95 ppm for 18 hours).G6PD levels were significantly lowered when the samples were dried under all the conditions except for fast cold drying (2 to 8 ℃ overnight and formaldehyde condition,0.30 to 0.38 ppm for 4 hours or 0.21 to 0.24 ppm for 18 hours).TSH and 17α-OHP levels were lowered obviously when the samples were dried under the conditions of humidity,UV and formaldehyde condition (TSH:0.32 to 0.52 ppm for 4 hours,0.38 to 0.45 ppm for 18 hours,17α-OHP:4.37 to 4.62 ppm for 4 hours,0.38 to 0.45 ppm for 18 hours).The results of Phe,G6PD,TSH and 17α-OHP were not statistically different with the control group when the samples were dried under the fast cold drying and 2 to 8 ℃ overnight.Conclusion The physical and chemical factors in the environment of DBS preparation should be related to the accuracy of neonatal disease screening closely.The necessary control factors including formaldehyde,ethanol,glacial acetic acid,ultraviolet irradiation,heat,humidity and decoration pollution may exhibit significant effects on the preparation of DBS.Fast cold drying and overnight at 2 to 8 ℃ could be available for DBS preparation.

"Practice and thinking on the construction of the""new ecology of the integration of medical service and aging care""in Wuxi" / 中华医院管理杂志

An introduction to the concept ofintegration of medical service and aging care,the strategic goals and policies in support of the new ecology of integration of medical service and aging care.The authors described specific practices in the development of laws and regulations,provision of various services and update means to build such an ecology in Wuxi,along with proposals to speed up the innovation and development.

Predominant D1 Receptors Involvement in the Over-expression of CART Peptides after Repeated Cocaine Administration

The aim of this study was to investigate the involvement of dopaminergic receptors (DR) in behavioral sensitization, as measured by locomotor activity, and the over-expression of cocaine- and amphetamine-regulated transcript (CART) peptides after repeated administration of cocaine in mice. Repeated administrations of cocaine induced behavioral sensitization and CART over-expression in mice. The levels of striatal CART mRNA were significantly increased on the 3rd day. CART peptides were over-expressed on the 5th day in the striata of behaviorally sensitized mice. A higher proportion of CART+ cells in the cocaine-treated mice were present in the nucleus accumbens (NAc) shell than in the dorsolateral (DL) part of caudate putamen (CP). The concomitant administration of both D1R and D2R antagonists, SCH 23390 (D1R selective) and raclopride (D2R selective), blocked cocaine induced-behavioral sensitization, CART over-expression, and cyclic adenosine 5'-monophosphate (cAMP)/protein kinase A (PKA)/phospho-cAMP response element-binding protein (pCREB) signal pathways. SCH 23390 more predominantly inhibited the locomotor activity, CART over-expression, pCREB and PKA activity than raclopride. Cocaine induced-behavioral sensitization was also attenuated in the both D1R and D2R knockout (KO) mice, respectively. CART over-expression and activated cAMP/PKA/pCREB signal pathways were inhibited in the D1R-KO mice, but not in the D2R-KO mice. It is suggested that behavioral sensitization, CART over-expression and activated cAMP/PKA/pCREB signal pathways induced by repeated administration of cocaine could be more predominantly mediated by D1R.

Effects of over-expression of ANXA10 gene on proliferation and apoptosis of hepatocellular carcinoma cell line HepG2 / 华中科技大学学报（医学）（英德文版）

The effects of over-expression of ANXA10 gene on proliferation and apoptosis of hepato-cellular carcinoma cell line HepG2 were elucidated. The human ANXA10 gene was subcloned into the lentiviral vector, PGC-FU, to generate the lentiviral expression vector, PGC-FU-ANXA10. The corrected ANXA10 was confirmed by endoenzyme digestion, and sequencing. Recombinant lentiviruses were produced by 293T cells following the co-transfection of PGC-FU-ANXA10 with the packaging plasmids pHelper1.0 and pHelper2.0. The resulting recombinant lentiviruses carrying ANXA10 were then used to infect human embryonic kidney epithelial cells, and lentiviral particles were produced. The ANXA10 expression in 293T cells was detected by using fluorescent microscope and Western blotting. HepG2 cells were infected, and divided into PGC-Fu-ANXA10 group, PGC-Fu group and HepG2 cell group. The changes of ANXA10 mRNA and protein expression were detected by using RT-PCR and Western blotting respectively. Flow cytometry and MTT assay were performed to examine the changes in cell apoptosis and proliferation respectively. The recombinant PGC-FU-ANXA10 vector was successfully constructed, the ANXA10 protein was detected by using Western blotting, and virus titer was 2×10(8) TU/mL. The recombinant lentiviruses were effectively infected into HepG2 cells in vitro and the infection efficiency was 70%. At 72 h after infection, the ANXA10 mRNA and protein expression levels in PGC-Fu-ANXA10 group were significantly higher than in PGC-Fu group and HepG2 cell group (P<0.05); the in vitro growth inhibition rate of HepG2 cells in PGC-Fu-ANXA10 group was 24.65%, significantly higher than that in PGC-Fu group and HepG2 cell group (P<0.05), but there was no significant difference between PGC-Fu group and HepG2 cell group; the apoptosis rate in PGC-Fu-ANXA10 group, PGC-Fu group and HepG2 cell group was (51.92±1.41)%, (19.00±1.12)% and (3.59±0.89)% respectively. The apoptosis rate in PGC-Fu-ANXA10 group was significantly higher than in PGC-Fu group and HepG2 cell group (P<0.05). The recombinant lentiviruses PGC-FU-ANXA10 were constructed successfully and infected into HepG2 cells. The overexpression of ANXA10 gene can significantly inhibit proliferation and promote apoptosis of HepG2 cells in vitro.

Diffusion tensor imaging of the prostate cancer / 中华放射学杂志

Objective To explore the diagnostic value of DTI for prostate cancer.Methods From October 2009 to December 2010,44 patients suspected of prostate cancer received MRI and DTI.The data of MRI and DTI were analyzed retrospectively.By histopathology,prostate cancer was proved in 16 patients,and benign prostatic hyperplasia ( BPH ) was proved in 28 patients.Differences in ADC and FA values between prostate cancer and BPH were compared by independent samples t test.Diagnostic accuracy of FA value and ADC value for prostate cancer was analyzed by using ROC curve,and the diagnostic threshold of FA value and ADC value for prostate cancer was determined.Results The mean FA value of the tumor regions and BPH were 0.308 +0.084 and 0.203 ±0.029,respectively.The mean ADC value of the tumor regions and BPH were (0.883 +0.192) × 10 -3 mm2/s and ( 1.408 ±0.130) × 10-3 mm2/s,respectively.There were statistically significant differences in ADC and FA values between tumor regions and BPH (t values were 4.833 and 10.779 respectively,P＜0.01).The ADC value area under curve of ROC was 0.996 (95％ CI was 0.984 to 1.007) ; the FA value area under curve of ROC was 0.904(95％ CI was 0.812 to 0.996) ; Combined the FA and ADC value area under curve of ROC is 0.996(95％ CI was 0.984to 1.007) ; Using the ADC value of 0.725 × 10 3 mm2/s as the ROC cut off point,the diagnostic sensitivity and specificity were 100.0％ and 96.0％,respectively; Using the FA value of 0.311as the ROC cut off point,the diagnostic sensitivity and specificity was 100.0％ and 68.7％,respectively.Conclusion DTI imaging can provide valuable information for prostate cancer diagnosis and differential diagnosis,and improve the diagnosis ability of prostate cancer.

Role of stress in depression insomnia and sleep characteristics of commonly used animal stress models / 药学学报

Depression and insomnia are intimately related. Depressed patients usually manifest sleep discontinuity and early awakening, reduced or no slow wave sleep (SWS) and shortened latency of rapid eye movement (REM) sleep. These sleep abnormalities are very similar to those caused by over activated hypothalamic-pituitary-adrenal (HPA) axis with stress. Therefore, the animal models developed by post-traumatic stress disorder or chronic unpredictable mild stress could be used to evaluate drugs which have effects of both anti-depression and improvement of sleep quality, and to provide a more reliable platform for further studis on the mechanisms of depression and accompanied insomnia. This review mainly focuses on the typical features of sleep disturbance of depression, possible pathophysiological mechanisms, establishment of animal stress models and analysis of their abnormal sleep characteristics.

Effects of over-expression of ANXA10 gene on proliferation and apoptosis of hepatocellular carcinoma cell line HepG2 / 华中科技大学学报（医学）（英德文版）

The effects of over-expression of ANXA10 gene on proliferation and apoptosis of hepato-cellular carcinoma cell line HepG2 were elucidated. The human ANXA10 gene was subcloned into the lentiviral vector, PGC-FU, to generate the lentiviral expression vector, PGC-FU-ANXA10. The corrected ANXA10 was confirmed by endoenzyme digestion, and sequencing. Recombinant lentiviruses were produced by 293T cells following the co-transfection of PGC-FU-ANXA10 with the packaging plasmids pHelper1.0 and pHelper2.0. The resulting recombinant lentiviruses carrying ANXA10 were then used to infect human embryonic kidney epithelial cells, and lentiviral particles were produced. The ANXA10 expression in 293T cells was detected by using fluorescent microscope and Western blotting. HepG2 cells were infected, and divided into PGC-Fu-ANXA10 group, PGC-Fu group and HepG2 cell group. The changes of ANXA10 mRNA and protein expression were detected by using RT-PCR and Western blotting respectively. Flow cytometry and MTT assay were performed to examine the changes in cell apoptosis and proliferation respectively. The recombinant PGC-FU-ANXA10 vector was successfully constructed, the ANXA10 protein was detected by using Western blotting, and virus titer was 2×10(8) TU/mL. The recombinant lentiviruses were effectively infected into HepG2 cells in vitro and the infection efficiency was 70%. At 72 h after infection, the ANXA10 mRNA and protein expression levels in PGC-Fu-ANXA10 group were significantly higher than in PGC-Fu group and HepG2 cell group (P<0.05); the in vitro growth inhibition rate of HepG2 cells in PGC-Fu-ANXA10 group was 24.65%, significantly higher than that in PGC-Fu group and HepG2 cell group (P<0.05), but there was no significant difference between PGC-Fu group and HepG2 cell group; the apoptosis rate in PGC-Fu-ANXA10 group, PGC-Fu group and HepG2 cell group was (51.92±1.41)%, (19.00±1.12)% and (3.59±0.89)% respectively. The apoptosis rate in PGC-Fu-ANXA10 group was significantly higher than in PGC-Fu group and HepG2 cell group (P<0.05). The recombinant lentiviruses PGC-FU-ANXA10 were constructed successfully and infected into HepG2 cells. The overexpression of ANXA10 gene can significantly inhibit proliferation and promote apoptosis of HepG2 cells in vitro.

Study on changes of neurological function and neuron apoptosis after intravenous administration of bone marrow stromal stem cells for treating permanent focal cerebral ischemia in rats / 中国病理生理杂志

AIM:To explore the survivorship and the mechanism of the intravenous administration of bone marrow stromal stem cells(BMSCs) for treating permanent focal cerebral ischemia in rats.METHODS:After purified,proliferated,and marked with BrdU,the BMSCs were injected intravenously into rats 1 d after focal cerebral ischemia.Modified neurological severity score(mNSS) was evaluated before and following 1,7,14 and 28 d after middle cerebral artery occlusion(MCAO).Rats were executed at 1,7,14 and 28 d after MCAO.Brain sections were stained with hematoxylin and eosin(HE) for determining the infarct volume.Slides were stained by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling(TUNEL) and immunostaining for cleaved caspase-3 method for apoptosis detection and mechanism exploration in situ.RESULTS:mNSS in BMSCs-transplanted group at 14th day and 28th day of MCAO was significantly lower than that in control group(P