BACKGROUND: Preliminary study has proved that the bone marrow-derived mesenchymal stem cells (MSCs) in a rat emphysema model produced by use of trypsin alone can "homing" to the lesioned lung tissues, and participate in the formation of pulmonary arteries to promote lung tissue repair. Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) play equally a powerful role in promoting angiogenesis. OBJECTIVE: To observe the influence of bFGF, VEGF and MSCs in regeneration of pulmonary capillary and pathological repair of pulmonary emphysema rats. METHODS: Except normal control group, the remaining 5 groups of rats were exposed to tobacco smoke and received a single intratracheally instillation of porcine pancreatic elastase to induce emphysema models. Following successful modeling, rats of bFGF group were intratracheally injected with 400 U bFGF and rats of VEGF group with 2 μg VEGF, once a week for three times. MSCs group was injected 1 mL suspension of 4×10~9/L MSCs into tail vein. MSCs+VEGF group was injected MSCs into tail vein and intratracheally injected VEGF (2 ug, three times) at the same time. Model control and normal control groups were intratracheally injected with equal volume of sodium chloride. Four weeks after treatment, arterial blood gas analysis was performed to observe pathological and morphological changes of lung tissues. CD34~+ expression in lung tissues was determined using immunohistochemistry method. RESULTS AND CONCLUSION: Compared with model control group, PaO_2 values dramatically increased in VEGF group (P <0.05), while other indices remained unchanged (P > 0.05); there were no obvious changes in each index in other groups (P >0.05). Gross and microscopic observations showed that, lung was smooth, pale pink, and elastic in normal control group, with uniform size of pulmonary alveoli on cross-section; pathological changes of chronic obstructive pulmonary emphysema existed in model control group, but improved in other 4 groups. Compared with model control group, mean pulmonary alveoli number and CD34~+ relative positive area dramatically increased in bFGF, VEGF, MSCs, MSCs+VEGF groups (P < 0.05), mean linear intercept and mean alveoli area were significantly reduced (P < 0.05). No significant difference was observed in each index among these 4 groups (P > 0.05). bFGF, VEGF and MSCs could improved the pathology of pulmonary emphysema models produced by tobacco smoking and intratracheally instillation of porcine pancreatic elastase. The possible mechanism of recovering the pulmonary emphysema is the proliferation of pulmonary capillary and enlargement of pulmonary artery, improved blood flow in the lung, improved ventilation/perfusion shunt, reduced pulmonary alveolus size and volume of the lung through self-compensation.

Objective To investigate the feasibility of using clustered regularly interspaced short palindromic repeats ( CRISPR)-mediated genome editing to downregulate the expression of programmed cell death protein 1 (PD-1) on primary T cells by using a lentivirus delivery system. Methods Lentivirus vec-tors pLentiCRISPR A1-A6 containing different PD-1 genomic DNA sequences as single guide RNA ( sgRNA) for Cas9 targeting were constructed individually. The lentivirus vectors were tranduced into primary CD4 T cells. Flow cytometry analysis was performed to detect the expression of PD-1 for evaluating the knockout ef-ficiency. Results The lentivirus vectors pLentiCRISPR A1-A6 carrying six different target sites were con-structed and respectively tranduced into primary CD4 T cells. The expression of PD-1 accompanied with the activation of T cells. Co-expression of CD25 and PD-1 was observed on activated T cells. All of the six sites could be targeted by Cas9, of which A2 and A6 sites were more efficient in knocking out the gene encoding PD-1 with a rate of 19% and 29%, respectively. Conclusion This study suggests that it is feasible to knock out the expression of PD-1 on primary T cells by using CRISPR.

Objective To explore the function of 5-Aza-CdR in B-cell acute lymphocytic leukemia cell line NALM-6 and its influence on the expression of microRNA (miRNA) in the cells. Methods NALM-6 was treated with different concentrations of 5-Aza-CdR. Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) test, and DNA methyltransferase (DNMT) mRNA expression level was detected by reverse transcription PCR (RT-PCR). The expression changes of miRNA were detected by miScript miRNA PCR Array chip in cells after methylation. Results NALM-6 cell growth was inhibited by different concentrations of 5-Aza-CdR processing time, reaching to the maximum inhibitory rate was (74.163 ±0.381) %. 5-Aza-CdR affected concentrations was inversely proportional with expression level of DNMT mRNA. After 1 000 μmol/L of 5-Aza-CdR was dealed with cell 72 h, the relative expression of DNMT-1 was reduced to 0.453 ±0.021, DNMT-3L was 0.003±0.001, DNMT-3B was 0.395±0.019. MiScript miRNA PCR array sieved out 3 miRNA (miR-184, miR-23a-3p, miR-34a-5p) associated with DNA methylation. Conclusions 5-Aza-CdR down regulates the expression of DNMT gene in NALM-6 cells, and inhibits the proliferation of cells. MiR-184, miR-23a-3p and miR-34a-5p are related to DNA methylation in the occurrence and development of B-cell acute lymphocytic leukemia.

Objective To evaluate the effect of two polymorphisms(rs761737 and rs2269058) of RNF8 and the interactions with cigarette smoking and alcohol consumption on sperm DNA fragment index (DFI)and primary male infertility .Methods Based on case-control design ,polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology was used to de-tect the genotype of rs761737 and rs2269058 in RNF8 between 332 primary male infertile patients (composed by 87 patients of azoospermia ,166 patients of oligoasthenozoospermia and 79 patients of normozoospermia ) and 329 controls ,and Sperm Chromatin Dispersion(SCD) assay was used to assess sperm DNA fragment index (DFI) .Results Genotype and allele frequencies distribution of rs761737 and rs2269058 between cases and controls had no statistically significant difference (P>0 .05) .Sperm DFI in infertile group (46 .2 ± 22 .3)% was significantly higher than that of control group (21 .4 ± 9 .2)% (P<0 .05) ,stratified analysis suggested that Sperm DFI in oligoasthenozoospermia group (50 .0 ± 22 .1)% was also significantly higher than that of normozoospermia group (38 .2 ± 20 .7)% .The statistic differences of Sperm DFI in individuals who carried different genotypes of rs 761737 and rs2269058 in oligoasthenozoospermia group and normozoospermia group had no statistically significant difference (P>0 .05) .There was an inter-action between RNF8 rs2269058 and Cigarettes smoking(P<0 .05 ,OR=2 .37 ,95% CI 1 .06-5 .27) .Conclusion Although RNF8 rs761737 and rs2269058 have no effects on primary male infertility and sperm DFI ,cigarettes smoking increase the risk of primary male infertility in individuals who carry RNF8 rs2269058 AC+AA genotype .Sperm DFI is an important test to assess sperm quali-ty ,it is vital to reveal the etiology of primary male infertility and provide therapy guidance to clinicians .

Objective To analyze the type and gene features of 11 unverified HAdVs in China. Methods We downloaded HAdV genome sequences of 67 strains with clear genotypes and the HAdV reference genome sequences of 7 strains from GenBank database, analyzed them together with eleven unverified HAdV gene sequences and classified the species of each sequence based on the genetic structure, sequence similarity and phylogenetic analysis. Results Phylogenetic analysis results predicted the classification of six sequences: two of them belonged to HAdV-B and two belonged to HAdV-D (the gene homology were 97.8% and 92.8% in comparison with reference gene respectively), the rest of two sequences were not conclusive. Multiple sequence alignment was used to predict the species of five HAdVs. All of them belonged to species HAdV-B, the sequence similarity compared with reference gene was 97.9%. Conclusion Genotyping of the unverified HAdV genome from a genetic point of view predicts that seven strains might belong to species HAdV-B, two strains might belong to species HAdV-D and two strains could not be classified. The seven HAdV-B strains might be suitable for the treatment of osteosarcoma as oncolytic adenovirus.

Objective To study the clinic effect and safety of MEAD chemotherapy regimen for adult patients with relapsed or refractory acute lymphocyte leukemia. Methods Between July 2006 and July 2009,twenty-two adult patients with relapsed or refractory acute lymphocyte leukemia received MEAD regimen (mitoxantrone 6 mg/d dl-3 iv drip,cytarabine 100 mg/d dl-5 iv drip,etoposide 100 mg/d dl-5 iv drip,dexmethasone 10 mg/d dl-8 iv drip). Results The complete remission (CR) rate of adult patients with relapsed or refractory acute lymphocyte leukemia was 31.8 %,the partial remission(PR) rate was 22.7 % and the overall response (OR) rate 54.5 %. The cumulitive CR rate was 50.0 %,and the PR rate 40.9 % after two times MEAD chemotherapy regimen. The main adverse effect was different level of myelosuppression,and other toxicity of vital organ was mild. Conclusion MEAD regimen is effective and can be tolerated for adult patients with relapsed or refractory acute lymphocyte leukemia,and its side effect is mild.

Objective To investigate the expression level and clinical significance of WT1 gene in acute luekemia (AL) patients.Methods WT1 gene level was detected by real time quantitative-polymerase chain reaction in acute myelogenous leukemia (AML) and in acute lymphocytic leukemia (ALL) patients.Then the expression levels of WT1 gene in different subtypes of AML were compared,and the correlation between gene expression and disease courses and prognosis were observed.Moreover,the relationship between disease courses and WT1 expression in patiens after receiving haemopoietic stem cell transplantation were analyzed.Results Among 66 cases,WT1 expression positive rate was 87.5 ％ (14/16) in AML and 76.0 ％ (38/50) in ALL.In AML,the expression level in M3 showed the lowest than that in any other subtypes (compared with M1,M2,M4,M5,P value was 0.040,0.007,0.006 and 0.01,respectively).The expression level of WT1 was closely correlated with leukemia disease courses.The expression level in complete remission (CR) group showed a significant lower expression level than that in non-remission group (P =0.018) and relapse group (P =0.003),and the re-increase of WT1 expression level could predict relapse as early as 1.5 months.Moreover,WT1 expression also showed an close relationship with prognosis of patients receiving haemopoietic stem cell transplantation.Patients whose WT1 was undetectable had a better prognosis than those with persistent expression,and increase again after becoming undetectable.Conclusion WT1 has a high expression level in AL,which can represent minimal residual disease.The expression level in M3 was lowest than that in different AML subtypes,and its expression level has a close correlation with clinical disease course and prognosis of AL.

Objective To investigate the effect of dexmedetomidine on postoperative cognitive function in elderly patients undergoing general anesthesia.Methods A total of 100 elderly patients undergoing elective surgery were enrolled.The patients were randomly divided into observation group and control group,with 50 patients in each group.The observation group was given dexmedetomidine + 0.9％ sodium chloride injection before induction anesthesia.The control group was given 0.9％ sodium chloride injection.The remaining anesthesia induction and maintenance time were same in the two groups.The cognitive function and postoperative cognitive dysfunction(POCD) were evaluated by MMSE scale.The blood pressure and heart rate before and after operation and at the end of the operation,the levels of serum tumor necrosis factor α(TNF-α) before operation,at the end of the operation and postoperative 3,7 days were measured.Results At postoperative 1 day and 3 days,the MMSE scores of the observation group were (24.6 ± 0.7) points and (27.2 ± 1.1) points,respectively,which were significantly higher than those of the control group [(22.4 ± 0.6) points,(27.2 ± 1.1) points,t =3.64,3.97,all P ＜ 0.05].And 6 cases in the observation group occurred cognitive dysfunction,which was significantly lower than 20 cases of the control group (x2 =5.38,P ＜0.05).At the end of the operation and postoperative 3 days,the serum levels of TNF-α in the observation group were (28.52 ± 3.73) ng/L,(28.82 ± 4.13) ng/L,respectively,which were significantly lower than those in the control group[(37.14 ± 3.62) ng/L,(38.27 ± 3.47) ng/L,t =5.21,4.89,all P ＜ 0.05].At the beginning of the operation,the systolic blood pressure,diastolic blood pressure,heart rate of the observation group were (124.53 ±3.84) mmHg,(79.32 ± 4.38) mmHg,(70.45 ± 5.32) times/min,respectively,which were significantly lower than those of the control group [(145.36 ± 4.72) mmHg,(93.17 ± 3.82) mmHg,(86.79 ± 4.26) times/min],the differences were statistically significant(t =5.83,4.97,4.58,all P ＜ 0.05).Conclusion In the elderly patients undergoing general anesthesia,the use of pumped dexmedetomidine has little effect on the postoperative cognitive function.The mechanism may be related to stabilizing the blood flow dynamics,decrease the high expression of TNF-α,which is worthy of clinical application.

Objective:To evaluate the safety and efficiency of a modified double-tract reconstruction procedure─proximal gastrectomy with piggyback interposed jejunal single-channel reconstruction (PJIR-STR) for early SiewertⅡ adenocarcinoma of esophagogastric junction (AEG).Method:Data of 8 SiewertⅡ AEG patients at Shanxi Tumor Hospital and undergoing PJIR-STR from May 2018 to Oct 2019 were retrospectively analyzed. The gastroesophageal reflux disease questionnaire (GerdQ) was used to score the patients at 3, 6, 12, and 18 months after surgery. The severity of postoperative reflux esophagitis was assessed by gastroscopy at 3, 6 months after surgery, using the Los Angeles Classification criteria.Result:All patients recovered well after surgery without serious complications. No obvious gastroesophageal reflux was observed in all patients at different periods (All of the GerdQ scores were <8 points.) The results of gastroscopy showed that 1 patient was diagnosed as grade B reflux esophagitis at 3, 6 months after surgery, which was responsive to conservative treatment, and the other 7 patients had no grade B or above reflux esophagitis.Conclusion:PJIR-STR is a feasible, safe reconstruction with excellent efficiency of dual anti-reflux for the SiewertⅡ AEG.

Objective To evaluate the application effect of the quality control circle(QCC) on shortening DNT in patients with cerebral ischemic stroke. Methods QCC activity group had been established and the reasons for the DNT extension in patients with cerebral ischemic stroke were analyzed so as to identify key improvement links and to formulate and to implement countermeasures. Fifteen cases with cerebral ischemic stroke in our hospital before the implementation of QCC from March to April in 2016 were selected as the control group, and 32 cases of patients with cerebral ischemic stroke after the implementation of QCC activities from May to August in 2016 were selected as the observation group. DNT in the two groups of thrombolytic cases were compared, and the more time consuming of emergency, blood test and signature communication were compared and optimized. Results The average DNT of patients with acute cerebral stroke in the observation group was significantly shorter than that in the control group, and the difference was statistically significant (P＜0. 05). There were 5 cases with more than 110min DNT in the control group and no case with more than 110min DNT in the observation group, and the difference was statistically significant (P＜0. 05). The average time consuming in emergency treatment, blood specimen test and signature communication in the observation group was significantly shorter than that in the control group, and the differences were statistically significant(P＜0. 05). Conclusion Through the implementation of QCC can effectively regulate the emergency treatment process of stroke and shorten the DNT in patients with cerebral ischemic diseases.