Effect of cold storage time on cellular viability of umbilical cord mesenchymal stem cells / 中国组织工程研究

BACKGROUND: With the in-depth basic research and clinical research on mesenchymal stem cells, mesenchymal stem cells have wide clinical prospects. However, little is reported on the temporary storage conditions and refusion time of mesenchymal stem cells. OBJECTIVE: To explore the effect of cold storage time on the cell mass and viability of umbilical cord mesenchymal stem cells.METHODS: Frozen-thawed and unfrozen umbilical cord mesenchymal stem cells were prepared and stored at 0-4 ℃. The cell viable cell rate, cell doubling time and colony forming efficiency were detected after 0, 6, 12, and 24 hours. RESULTS AND CONCLUSION: Unfrozen cells could maintain the cell biological activity at 0-4 ℃until dead cells appeared with the presence of decreased cell viability at 12 hours after storage. Frozen-thawed cells were unable to be stored at 0-4 ℃ for a long time, and cells began to die and the cell viability was weakened at 6 hours after storage. These findings indicate that umbilical cord mesenchymal stem cells should be injected into patients within 6 or 12 hours after preparation, to ensure the best therapeutic effects. If there is a longer transport distance, it is preferred to use unfrozen mesenchymal stem cells.

[Effects of Buzhong Yiqi decoction on expression of Bad, NF-κB, caspase-9, Survivin, and mTOR in nude mice with A549/DDP transplantation tumors].

This study was aimed to explore the effects of Buzhong Yiqi decoction on the expression levels of Bad, NF-κB, caspase-9, Survivin, and mTOR in nude mice with A549/DDP transplantation tumors.Sixty BALB/C mice were randomly divided into blank control group, tumor-bearing control group, cisplatin group and Buzhong Yiqi decoction of high, medium and low doses+cisplatin groups (hereinafter referred to as the high,medium and low combined groups). A549/DDP cells (concentration of 5×106 cells/mL)were cultured and inoculated in various groups, then the tumor-forming situations were observed. Corresponding treatment was given in all groups. Fourteen days later, immunohistochemistry and Real-time PCR methods were used to detect the expression levels of Bad, NF-κB, caspase-9, Survivin, mTOR protein and mRNA in tumors.Results showed that Buzhong Yiqi decoction combined with cisplatin could reduce the volume of transplanted tumors, and there was significant difference between medium combined group and high combined group(P<0.05). As compared with the tumor-bearing control group, the expression levels of Bad, NF-κB, Survivin and mTOR were significantly reduced in medium and high combined groups(P<0.05); the protein and mRNA expression levels of caspase-9 were gradually increased in medium combined and high combined groups(P<0.05), with statistical difference with tumor-bearing control group(P<0.05). There were statistical difference in mRNA expression of Bad, NF-κB and caspase-9 between medium combined group, high combined group and cisplatin group, low-combined group, tumor-bearing control group(P<0.05), but there was no statistical difference between cisplatin group, low-combined group, and tumor-bearing control group. In addition, there was no statistical difference between medium combined group and high combined group in protein and mRNA expression levels of various factors. Experimental results showed that Buzhong Yiqi decoction combined with cisplatin can inhibit the growth of A549/DDP transplanted tumors, and the mechanism may be associated with regulating Bad, NF-κB, caspase-9, Survivin, and mTOR levels as well as promoting apoptosis.

3D visualization assisted proximal femoral nail anti-rotation surgery in the treatment of elderly intertrochanteric fracture / 局解手术学杂志

Objective To investigate the application of 3 D visualization (3 DV) assisted proximal femoral nail anti-rotation (PFNA) surgery in the treatment of elderly intertrochanteric fracture (ITF).Methods Totally 11 cases of elderly unstable ITF were treated with 3DV assisted PFNA fixation surgery.All these patients were examined with multi-row spiral CT volume scan,and the anatomical data of Dicom was reconstructed through M3 D software.Computer aided design (CAD) was performed based on 3 D reconstruction data,including anatomical data measurement,fracture reduction simulation,and proximal femoral head nail (PFNA) implant parameter design.And the CAD data were showed by S-3DV system during operation to guide the operation.Time of operation,intra-operative blood lose,post-operative drainage volume,hospital stay time and Harris hip score were recorded and compared to the non-3DV assisted PFNA procedure.Results In 3 DV-PFNA surgery,there was no significant difference between the preoperative planned average PFNA size and the implanted average PFNA size (P ＞ 0.05).Compared to the non-3DV-PFNA surgery,3 DV-PFNA surgery has less intra-operative blood lose and post-operative drainage volume,shorter time of preoperative traction reduction and operation (P ＜ 0.05).Harris hip function score of 3 DV-PFNA surgery was better than the conventional surgery 1 week after operation,and there was no significant difference in VAS score I month after operation.The 11 cases treated with 3DV-PFNA technology were healed well,and the rate of complications was lower than the conventional surgery.Conclusion 3DV technique can show accuracy CAD model of ITF,which may play an important role to improve efficiency and accuracy of PFNA surgery.

Identification and phylogenetic analysis for Achromobacter xylose Oxidas in porcine pneumonia / 中国人兽共患病学报

To identify the pathogen of an outbreak of pneumouia in pig-farm,we used biochemical identification,pathoge nicity test in mice,PCR identification by 16S rRNA,and phylogenetic tree analysis for the identification.Results showed that the pathogen was an Achromobacter xylosoxidans with certain virulence and named TLSY-1.This TLSY-1 was the most simi lar to KF879922.1,and affinity rate were 99.9％ by homology analysis.This result identified that TLSY-1 is a new pneumonia pathogens in piglets.

Baseline Naive CD4+ T-cell Level Predicting Immune Reconstitution in Treated HIV-infected Late Presenters / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Among HIV-infected patients initiating antiretroviral therapy (ART), early changes in CD4+ T-cell subsets are well described. However, HIV-infected late presenters initiating treatment present with a suboptimal CD4+ T-cell reconstitution and remain at a higher risk for AIDS and non-AIDS events. Therefore, factors associated with CD4+ T-cell reconstitution need to be determined in this population, which will allow designing effective immunotherapeutic strategies.</p><p><b>METHODS</b>Thirty-one adult patients with baseline CD4+ T-cell count <350 cells/mm3 exhibiting viral suppression after ART initiation were followed in the HIV/AIDS research center of Peking Union Medical College Hospital in Beijing, China, from October 2002 to September 2013. Changes in T-cell subsets and associated determinants were measured.</p><p><b>RESULTS</b>Median baseline CD4+ T-cell count was 70 cells/mm3. We found a biphasic reconstitution of T-cell subsets and immune activation: a rapid change during the first 6 months followed by a more gradual change over the subsequent 8 years. Baseline CD4+ T-cell count >200 cells/mm3 in comparison to CD4+ T-cell count ≤200 cells/mm3 was associated with more complete immune Reconstitution (77.8% vs. 27.3% respectively; P = 0.017) and normalized CD4/CD8 ratio. We showed that the baseline percentage of naive CD4+ T-cell was a predictive marker for complete immune reconstitution (area under receiver operating characteristic curve 0.907), and 12.4% as cutoff value had a sensitivity of 84.6% and a specificity of 88.2%.</p><p><b>CONCLUSIONS</b>Baseline naive CD4+ T-cell percentage may serve as a predictive marker for optimal immune reconstitution during long-term therapy. Such study findings suggest that increasing thymic output should represent an avenue to improve patients who are diagnosed late in the course of infection.</p>

Prevalence of thrombocytopenia among Chinese adult antiretroviral-naïve HIV-positive patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The prevalence of thrombocytopenia among Chinese antiretroviral therapy (ART)-naïve HIV-infected adults has not been well-described. The aim of this study was to investigate the prevalence and associated risk factors of thrombocytopenia among Chinese ART-naïve HIV-infected adults.</p><p><b>METHODS</b>We performed a cross-sectional study of Chinese adult ART-naïve HIV-infected patients from September 2005 through August 2014. Socio-demographic variables and laboratory results including platelets, CD4+ cell count, and viral load were obtained from medical records. Factors and outcomes associated with thrombocytopenia were assessed using logistic regression.</p><p><b>RESULTS</b>A total of 1730 adult ART-naïve HIV-infected patients was included. The mean age was 38 years. The prevalence of thrombocytopenia was 4.5%. There were significant differences in the prevalence of thrombocytopenia between patients <30 years of age (2.8%) and 30-39 years (4.0%) compared with patients greater than 50 years (7.0%) (P = 0.006 and P = 0.044, respectively). The prevalence of thrombocytopenia was also significantly different between patients with CD4+ counts of 200-349 cells/mm 3 (3.3%) and >350 cells/mm 3 (2.8%) compared with patients with CD4+ counts of 50-199 cells/mm 3 (7.1%) (P = 0.002 and P = 0.005, respectively). The prevalence of thrombocytopenia was significantly different by hepatitis C virus antibody (HCV-Ab) seropositivity (10.2% for HCV-Ab positive vs. 3.9% for HCV-Ab negative, P = 0.001). We observed differences in prevalence of thrombocytopenia by mode of transmission of HIV infection: Blood transmission (10.7%) versus men who have sex with men (3.9%) (P = 0.002) and versus heterosexual transmission (3.9%) (P = 0.001). In binary logistic regression analyses, age ≥ 50 years, HCV-Ab positivity and having a CD4+ cell count of 50-199 cells/mm 3 were significantly associated with thrombocytopenia with adjusted odds ratio of 2.482 (95% confidence interval [CI]: 1.167, 5.281, P = 0.018), 2.091 (95% CI: 1.078, 4.055, P = 0.029) and 2.259 (95% CI: 1.028, 4.962, P = 0.042), respectively.</p><p><b>CONCLUSIONS</b>Thrombocytopenia is not common among adult ART-naïve HIV-infected patients in China. Older age (age over 50 years), HCV-Ab positivity and lower CD4+ cell count are associated with an increased risk of thrombocytopenia. Therefore, early diagnosis and treatment of thrombocytopenia in these patients are necessary.</p>

[Buzhong yiqi decoction containing serum reversed resistance of A549/DDP to cisplatin and its effect on the expression of survivin: an experimental research ].

OBJECTIVE: To analyze the reversal effect of Buzhong Yiqi Decoction (BYD) on multidrug resistance of human adenocarcinoma of lung cell line A549/DDP, and to study its effect on the expression of survivin by using serum pharmacological methods in vitro. Methods Totally 24 SD rats were divided into the high, medium and low dose groups, and the blank control group by randomized controlled method. The high dose BYD containing serum (1. 134 g/mL, 2 mL), the middle dose BYD containing serum (0.576 g/mL, 2 mL), and the low dose BYD containing serum (0.284 g/mL, 2 mL) were prepared. The inhibitory effects of different dose and concentrations BYD on the proliferation of A549 and A549/DDP cells were detected by MTT assay, and the drug resistance reversal fold was calculated. The expression of Survivin in the two cell strains were detected respectively by immunohistochemical assay, Western blot, and immunofluorescence method. RESULTS: BYD containing serum showed obvious inhibitory effect on the growth of A549 and 549/DDP. The inhibition rates of 10% dose groups were higher than those of 5% dose groups. Besides, it gradually increased along with increased concentrations. Compared with 10% blank control group, the inhibition rate increased in 10% middle and low dose groups (P <0.05). After acted with 10% middle dose BYD containing serum, IC50, of A549 and A549/DDP were both reduced (P <0.05), reversal fold (RF) both increased. Its reversal ratio on A549/DDP cells was 2. 46, decreasing the resistance of A549/DDP to DDP. Compared with A549 in the same group, the expression of Survivin was detected to decrease by immunocytochemical assay, Western blot, and immunofluorescence method (P<0.05). Compared with 10% blank control group, the inhibition rate decreased in 10% middle dose group (P <0. 05). CONCLUSIONS: 10% middle dose BYD containing serum could significantly inhibit the apoptosis of A549 and A549/DDP. Besides, it could moderately reverse the multidrug resistance of A549/DDP cells to DDP possibly through reducing the intracellular expression of Survivin and enhancing the sensitivity 549/DDP to chemotherapeutics.

[Effect of Buzhong Yiqi decoction on PI3K and AKT in spleen, stomach and lung of nude mice with lung adenocarcinoma transplantation tumor].

OBJECTIVE: To explore the effect of Buzhong Yiqi decoction on PI3K/AKT signaling pathway in spleen, stomach and lung of nude mice with lung adenocarcinoma transplantation tumor. METHOD: Totally 60 nude mice were randomly divided into the blank control group, the tumor-bearing control group, the cisplatin group, the low-dose Buzhong Yiqi decoction group, the middle-dose Buzhong Yiqi decoction group and the high-dose Buzhong Yiqi decoction group. After the corresponding interventions, efforts were made to measure the transplanted tumor volume and calculate the tumor inhibiting rate. The immunohistochemical method and real time PCR were used to detect the expression of PI3K and AKT level in nude mice spleen, stomach and lung. RESULT: Buzhong Yiqi decoction of different concentrations combined with cisplatin could inhibit the growth of the transplanted tumor, with the strongest inhibitory effect in the middle-dose Buzhong Yiqi decoction group and the high-dose Buzhong Yiqi decoction group. All of the expressions of PI3K and AKT protein and gene in the spleen, stomach and lung increased, with the most significant increase in the tumor-bearing group. Along with the increase of the concentration of cisplatin and Buzhong Yiqi decoction, the expressions of PI3K and AKT gradually reduced. Compared with the tumor-bearing control group, there were statistical differences in spleen and stomach tissues (P < 0.05). Compared with the cisplatin group, the middle-dose Buzhong Yiqi decoction group and the high-dose Buzhong Yiqi decoction group showed statistical differences (P < 0.05), but without statistical difference compared with the blank control group. CONCLUSION: Among nude mice with lung adenocarcinoma transplantation tumor, the PI3K and AKT protein and gene expressions in spleen, stomach and lung tissues increased, which might indicated the effect of cisplatin and Buzhong Yiqi decoction in reducing PI3K and AKT expressions and the relations between the reduction degree and the concentrations of Buzhong Yiqi decoction. Cisplatin combined with Buzhong Yiqi decoction could decrease the PI3K and AKT protein and gene expression in spleen, stomach and lung, and make the pathway closer to normal, so as to protect the functions of spleen, stomach and lung, there may be target spots of Buzhong Yiqi decoction in PI3K/AKT signal pathway.

Hepatitis B virus/human immunodeficiency virus coinfection: interaction among human immunodeficiency virus infection, chronic hepatitis B virus infection, and host immunity / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>This review discusses progress in the studies of hepatitis B virus (HBV)/human immunodeficiency virus (HIV) coinfection and focuses on the interaction among HIV infection, chronic HBV infection, and host immunity.</p><p><b>DATA SOURCES</b>Data and studies published mainly from 2008 to 2011 were selected using PubMed.</p><p><b>STUDY SELECTION</b>Original articles and critical reviews concerning HBV/HIV coinfection and HBV and HIV pathogenesis were selected.</p><p><b>RESULTS</b>HIV may accelerate HBV progression by lowering CD4 count, weakening HBV-specific immunity, "enriching" HBV mutants, causing immune activation, etc. On the other hand, HBV may enhance HIV replication by activating HIV long terminal repeat (LTR) with X protein (HBX) and cause immune activation in synergy with HIV. Paradoxically, HBV may also inhibit HIV dissemination via dendritic cells.</p><p><b>CONCLUSIONS</b>The interaction among HIV, HBV, and host immunity remains poorly understood. Further research is warranted to elucidate the detailed molecular mechanisms and to translate these mechanisms into clinical practice.</p>