Objective:To explore the expression patterns of inhibitor of differentiation(ID)family in patients with chronic myeloid leukemia(CML)and analyze their clinical implications.Methods:Quantitative PCR and quantitative methylation-specific PCR were conducted to de-tect the transcript levels of ID2/ID3/ID4 and the methylation levels of ID4 in the bone marrow mononuclear cells of non-hematological ma-lignancies(acting as controls)and patients with CML treated at The Affiliated People's Hospital of Jiangsu University from January 2010 to December 2017.The clinical implications of ID family alterations were further analyzed.Results:ID2 and ID3 expression was significantly up regulated(P<0.001 and P<0.05,respectively),whereas ID4 expression was markedly down regulated in patients with CML(P<0.01).The re-ceiver operating characteristic curve demonstrated that the ID2 transcript level is a potential biomarker for distinguishing CML from controls(AUC=0.895,P<0.001).The frequency of ID4 promoter methylation in patients with CML was drastically higher than that in the controls(P=0.001).Moreover,ID4 methylation was negatively correlated with ID4 expression in patients with CML(r=-0.424,P=0.002).Clinically,CML with high ID2 expression occurred more frequently in males(P=0.040).Patients with low ID4 expression or high ID4 methylation showed a markedly higher frequency of an accelerated phase/blast crisis(P=0.003 and P<0.001,respectively).In addition,patients with CML in an accelerated phase/blast crisis exhibited markedly lower ID4 expression and higher ID4 methylation levels than those in the chronic phase(both P<0.001).Furthermore,univariate and multiple Logistic regression analyses revealed that the ID4 methylation level was an inde-pendent risk factor for CML progression(P=0.007).Conclusions:The ID family was differentially expressed in patients with CML;specifically,ID2 and ID3 expression was significantly increased,whereas ID4 expression was markedly decreased and correlated with ID4 promoter hy-permethylation.Hence,ID4 expression and methylation are confirmed to be associated with CML progression,and ID4 methylation could be an independent risk factor for CML progression.

Autologous ozonized blood transfusion(AOBT) is a therapy of re-transfusion of 100-200 mL of autologous blood after shaking and agitation with appropriate amount of oxygen-ozone in vitro. The oxidation of blood through the strong oxidation of ozone can enhance the non-specific immune response of the body, regulate the internal environment and promote health. This therapy has been increasingly applied in clinical practice, while no unified standard for the operation process in terms of ozone concentration, treatment frequency and treatment course had been established. This operation process of AOBT is primarily explored in order to standardize the operation process and ensure its safety and efficacy.

Objective:To construct 2019 novel coronavirus (2019-nCoV) 614D and 614G pseudovirus by HIV lentivirus packaging system and explore their biological specificity.Methods:The recombinant expression plasmids pCDNA3.1-614D and pCDNA3.1-614G were transiently cotransfected with psPAX2 and pLenti CMV Puro LUC into 293T cells respectively. After 72 hours, the supernatant was collected and ultracentrifuged with 20% sucrose cushion. The titer, morphology, protein expression and neutralizing activity of pseudovirus were determined.Results:S protein specific fluorescence was detected by indirect immunofluorescence test, Western blot analysis showed S protein was expressed, and the spike of pseudovirus was observed under transmission electron microscope. The titers of pseudovirus 614D and 614G were 1.12×10 4 and 2.52×10 4 TCID 50/ml, respectively. The pseudovirus 614D and 614G could be neutralized by S rabbit polyclonal antibody, indicating that the pseudovirus has high specificity. Conclusions:In this study, 2019-nCoV 614D and 614G pseudovirus was successfully constructed, which laid the foundation for the establishment of in vitro neutralizing antibody detection platform based on pseudovirus.

Objective To explore the erectile function preservational mechanism of Non-transecting urethroplasty(NTU) for posterior urethral stricture.Methods From May 2012 to September 2016,62 patients with posterior urethral stricture,who were treated with NTU,were enrolled in this study.The mean age was 37.5 years old,ranging 18-48 years old.The causes were pelvic fracture urethral injury in 53 cases and iatrogenic injury in 9 cases.Preoperative urethrography and urethroscopy revealed the strictures located in posterior urethra,which was at the distal of verumontanum.The mean length of stricture was 2.1 cm,ranging 0.5-2.5 cm.The average period between trauma and surgery was 6.4 months,ranging 3 months-2 years.All patients had no previous history of urethroplasty.Their sexual hormones were in normal level.Among those patients,the IIEF-5 scores were more than 12 and number of events during NPT test were more than twice.Finally,43 cases were underwent NTU and 19 cases accepted inferior pubectomy (IP)+ NTU.All patients had a general anesthesia.The bulbar urethra was mobilized dorsally from the tunica albuginea of the corpora cavernosa and then extended proximally up toward the perineal membrane.Scar tissue surrounding the urethra was excised and inferior pubectomy (IP) was performed as a supplemental technique to keep the suturing position without tension.The ventral hemi-circumference was then sutured with interrupted 4-0 polyglycolic sutures with tension-free anastomosis.The 18-Fr indwelling catheter was inserted.Result Average follow-up was 20.2 months,ranged from 12 to 36 months.In NTU group,NPT test revealed no significant difference in number of events (2.7 ± 0.7 vs.3.0 ± 1.0,P ＞ 0.05),duration of best episode [(16.4 ± 3.5) min vs.(16.4 ± 3.8) min,P ＞ 0.05)] or tip rigidity [(31.2 ± 4.7) ％ vs.(30.8 ± 3.5) ％,P ＞ 0.05)] between pre-and post-operation,respectively.The IIEF-5 score (19.7 ± 1.9 vs.20.4±2.1,P＜0.05)and Qmax[(8.7 ±4.0)ml/s vs.(25.5 ±4.7)ml/s,P＜0.05)] increased significant pre-and post-operation,respectively.In IP + NTU group,Qmax [(8.4 ± 4.4) ml/s vs.(23.1 ± 3.5)ml/s,P ＜ 0.05)] increased significant pre and post operation.The NPT test revealed slight decrease in number of events(2.3 ± 0.6 vs.1.6 ± 1.0,P ＜ 0.05),duration of best episode [(15.6 ± 2.4) min vs.(14.5±2.4)min,P＜0.05)] or tip rigidity [(29.8±3.0)％ vs.(25.6 ±7.1)％,P＜0.05)] between pre-and post-operation,respectively.However,the IIEF-5 scores (17.3 ± 1.6 vs.16.5 ± 2.1,P ＜ 0.05) didn't show significant difference pre-and post-operation.Stricture recurrence occurred in 3 patients,the success rate was 95.2％ (59/62) during 12 months following.Conclusion NTU is not only a safe and promising procedure for posterior urethral stricture less than 2.5cm,but also a new minimally invasive approach to preserve erectile function.

Objective:To investigate the functional status of dendritic cells(DCs)in oral leukoplakia(OLK)tissues.Methods:The expression of DC-specific markers CD1 a,CD209,CD1 23 and CD83 in 20 cases of OLK with abnormal dysplasia,1 0 with simple dys-plasia and 1 0 of normal oral mucosa tissues was examined by immunohistochemistry.Results:CD1 a and CD209 positive DCs were found in all cases.More CD1 a positive Langehans cells(LCs)in lamina propria were found in OLK with abnormal dysplasia than in normal o-ral mucosa and OLK with simple dysplasia(P <0.01 ).A great mount of CD209 positive stromal DCs were recruited in OLK.There was no CD83 positive and CD1 23 positive cell in normal oral mucosa,however,CD83 positive mature DCs and CD1 23 positive plasmacytoid DCs(PDCs)were observed in OLK(P <0.01 ).Conclusion:OLK is characterized by the recruitment of different subsets of DCs,the different DC subsets may play an important role in the development of OLK.

Objective To explore the role of portal venous pressure changes in the liver dysfunction caused by hepatic congestion after extended liver resection.Methods The experimental study was adopted.According to the random number table,90 Sprague-Dawley rats were divided into 3 groups,30 in each group:30 rats in the non-congestion group received 70％ of liver resection (median lobe + left lobe),30 rats in the congestion group received 70％ of liver resection (median lobe + left lobe) with whole caudal lobe congestion by ligation of veins and 30 rats in the congestion + splenectomy group received 70％ of liver resection (median lobe + left lobe) with whole caudal lobe congestion by ligation of veins and splenectomy.(1) Twenty rats in each group were used to make postoperative survival analysis.Ten rats in each group were used for related experiments.The portal venous pressures (PVPs) of 5 rats in each group were detected at postoperative 12 hours and 24 hours,and then blood and liver specimens were collected.(2) PVP changes were detected at postoperative 12 hours and 24 hours.(3) Clinical and biochemical test:level of total bilirubin (TBil) was tested at postoperative 12 hours and 24 hours.(4) Pathological examination:liver pathological damage was detected by HE staining.(5) The expression of CD68 macrophagocyte was detected by immunohistochemical staining.(6) The relative expressions of Cleaved Casepase-3 and hypoxia inducible factor-1α (HIF-1α) proteins at postoperative 24 hours were detected by Westein blot.(7) The relative expressions of mRNA of vascular regulation related genes (ET-1/eNOS) and inflammatory factors (TNF-α and IL-6) were detected by real-time polymerase chain reaction (RT-PCR).(8)The hyaluronic acid (HA) was measured by enzyme-linked immuno-sorbent assay (ELISA).Measurement data with normal distribution were represented as （x） ± s.Comparison among 3 groups was done using the ANOVA,and pairwise comparison was done by the LSD test.The postoperative 5-day survival curve was drawn by the KaplanMeier method,and the survival was compared using the Log-rank test.Results (1) Survival analysis:5-day survival rate in the non-congestion group,congestion group and congestion + splenectomy group were respectively 75％,10％ and 55％,with a statistically significant difference among the 3 groups (x2=18.21,P ＜0.05).(2)Changes of PVPs and TBil:levels of PVP and TBil in the non-congestion group,congestion group and congestion + splenectomy group were respectively (15.77 ±0.67)cmH2O,(18.33 ±0.28) cmH2O,(14.87 ± 0.58) cmH2O,(1.48 ±0.10)μmol/L,(1.76±0.15) μ mol/L,(1.62 ±0.11) μmol/L at postoperative 12 hours and (13.49 ± 0.45) cmH2 O,(16.96 ± 0.82) cmH2 O,(15.69 ± 0.85) cmH2 O,(1.47 ± 0.11) μmol/L,(1.94 ± 0.07) μmol/L,(1.67 ± 0.11) μmol/L at postoperative 24 hours,showing statistically significant differences among 3 groups (F =56.53,29.01,6.81,27.85,P ＜ 0.05).(3) Results of pathological examination:compared with noncongestion group,there were a lot of vacuolar cells with degeneration appearing in non-congestion liver tissues,severe liver cell swelling and hepatic sinus congestion in the congestion group at postoperative 24 hours.Compared with congestion group,vacuolar degeneration appearing in non-congestion liver tissues have some improvement in the congestion + splenectomy group.(4) Immunohistochemical staining:compared with non-congestion group and congestion + splenectomy group,the positive CD68 marked macrophages in the congestion group were increased at postoperative 24 hours.(5) Western blot assay:the relative expressions of Cleaved Casepase-3 and HIF-1α proteins in the non-congestion group,congestion group and congestion + splenectomy group were 0.63 ± 0.05,1.17 ± O.18,0.95 ± 0.17 and 0.63 ± 0.14,1.48 ± 0.08,1.13 ± 0.17,respectively,showing statistically significant differences among 3 groups (F =17.42,50.58,P ＜ 0.05).(6) Results of RT-PCR:the relative expression of mRNA of ET-1/eNOS in the non-congestion group,congestion group and congestion + splenectomy group was respectively 1.01 ± 0.63,2.09 ± 0.27,0.82 ± 0.12 at postoperative 12 hours and 0.73 ± 0.17,2.16 ± 0.94,0.80 ± 0.24 at postoperative 24 hours,showing statistically significant differences among 3 groups (F =62.91,10.65,P ＜0.05).The relative expression of mRNA of TNF-α in the non-congestion group,congestion group and congestion + splenectomy group was respectively 0.99 ± 0.08,127.80 ± 13.15,7.34 ± 1.56 at postoperative 12 hours and 0.99 ± 0.06,116.62 ± 13.32,58.62 ± 12.12 at postoperative 24 hours,showing statistically significant differences among 3 groups (F =436.77,154.54,P ＜ 0.05).The relative expression of mRNA of IL-6 in the non-congestion group,congestion group and congestion + splenectomy group was respectively 0.98 ±0.06,1.87 ±0.34,1.54 ±0.15 at postoperative 12 hours and 0.99 ±0.05,2.02 ±0.27,1.51 ±0.11at postoperative 24 hours,with statistically significant differences among 3 groups (F =22.08,46.71,P ＜ 0.05).(7) Results of ELISA:the level of HA in the non-congestion group,congestion group and congestion + splenectomy group was respectively (149 ± 9) ng/L,(200 ± 19) ng/L,(174 ± 9) ng/L at postoperative 12 hours and (136 ± 16) ng/L,(202 ± 13) ng/L,(91 ± 11) ng/L at postoperative 24 hours,with statistically significant differences among 3 groups (F =19.23,34.68,P＜0.05).Conclusions On the basis of extended liver resection,a wide range of liver congestion through increasing PVP causes hepatic microcirculation disorders,hypoxia,inflammation,vacuoles degeneration cells,increased cells apoptosis,aggravated damage of liver function and increased mortality of rats.Splenectomy could reduce PVP and then improve the liver tissues damage caused by liver congestion,meanwhile,increase the survival rate of rats.

OBJECTIVETo understand the toxic effects of manganese exposure on health by measurement of manganese concentration and occupational health inspection among workers in an enterprise exposed to manganese, and to provide a basis for occupational disease control.

METHODSThe air concentrations of manganese in 12 workplaces where workers often stayed were measured by typical sampling; occupational health inspection was performed among a total of 538 workers in the enterprise by cluster sampling.

RESULTSThe average concentrations of manganese in the workplace from 2011 to 2013 were 0.179 mg/m(3), 0.122 mg/m(3), and 0.082 5 mg/m(3), respectively, indicating a significant decrease in manganese level within the three years by rank-sum test (P < 0.05). The results of health inspection showed that 165 (30.7%) out of 538 workers had abnormal indices, and the incidence of autonomic nervous system abnormalities reached 32.7% (176/538). There was no correlation between manganese concentration in the workplace and the incidence of abnormal indices in health inspection, while the manganese concentration in the workplace was positively correlated with the incidence of autonomic nervous system symptoms (r = 0.718, P < 0.05).

CONCLUSIONThe manganese concentration in the workplace is positively correlated with the incidence of autonomic nervous system symptoms. Early exposure to a low level of manganese can induce functional changes and potential damages in the nervous system.

Humans ; Incidence ; Manganese ; analysis ; Nervous System ; drug effects ; physiopathology ; Occupational Diseases ; chemically induced ; epidemiology ; Occupational Exposure ; analysis ; Occupational Health ; standards ; Workplace

ObjectiveTo investigate the immunological improvement effects of different doses of ursodeoxycholic acid (UDCA) on patients with different stages of primary biliary cirrhosis (PBC) and enhance the understanding of the roles of the immune system in the disease, and to provide evidence for the standardized clinical treatment of PBC. MethodsOne hundred and eighty patients with PBC who were admitted to our hospital from March 2012 to Janurary 2014 were enrolled and equally divided into three classes according to the stage of PBC: early stage, cirrhotic stage, and poor biochemical response stage. Patients in each class were equally divided into three groups according to the dose of UDCA: 8-10, 13-15, and 20-25 mg·kg-1·d-1. The general information, clinical symptoms, biochemical indices, and changes in T lymphocyte subsets and cytokines in peripheral blood after the treatment with different doses of UDCA were analyzed. Comparison of continuous data was performed by t test, and comparison of categorical data was performed by χ2 test. ResultsIn patients with early-stage PBC who were treated with 13-15 mg·kg-1·d-1 UDCA, the percentage of CD3+CD4+ T cells, CD4+/CD8+ ratio, and expression of interferon-gamma were significantly reduced after treatment (54.8%±11.6% vs 34.7%±7.7 %, t=6.5, P＜0.05; 2.3±1.0 vs 1.6±0.6, t=2.7, P＜0.05; 33.0±12.3 vs 23.7±7.2 ng/L, t=2.9, P＜0.05), while the secretion of interleukin-4 was significantly increased after treatment (29.0±4.6 vs 38.5±7.1 ng/L, t=5.0, P＜0.05). ConclusionThe UDCA with a dose of 13-15 mg·kg-1·d-1 can substantially improve the immune status in patients with early-stage PBC. The application of UDCA should be standardized in order to achieve the desired response.

Objective To explore the effect of airway humidification on lung injury as a result of mechanical ventilation with different tidal volume(VT). Methods Twenty-four male Japanese white rabbits were randomly divided into four groups:low VT with airway humidification group,high VT with airway humidification group,low VT and high VT group without humidification,with 6 rabbits in each group. Mechanical ventilation was started after intubation and lasted for 6 hours. Low VT denoted 8 mL/kg,while high VT was 16 mL/kg,fraction of inspired oxygen (FiO2)denoted 0.40,positive end-expiratory pressure(PEEP)was 0. Temperature at Y piece of circuit in airway humidification groups was monitored and controlled at 40℃. Arterial blood gas analysis,including pH value,arterial partial pressure of oxygen(PaO2),arterial partial pressure of carbon dioxide(PaCO2),lung mechanics indexes, including peak airway pressure(Ppeak)and airway resistance(Raw),and lung compliance was measured at 0,2,4, 6 hours of mechanical ventilation. The levels of tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)in plasma and bronchoalveolar lavage fluid(BALF)were determined by enzyme linked immunosorbent assay(ELISA). The animals were sacrificed at the end of mechanical ventilation. The wet to dry(W/D)ratio of lung tissues was calculated. Histopathologic changes in the lung tissueies were observed with microscope,and lung injury score was calculated. Scanning and transmission electron microscopies were used to examine the integrity of the airway cilia and the tracheal epithelium. Results Compared with low VT group,pH value in high VT group was significantly increased,PaCO2 was significantly lowered,and no difference in PaO2 was found. Ppeak,Raw,and lung compliance were significantly increased during mechanical ventilation. There were no significant differences in blood gas analysis and lung mechanics indexes between low VT with airway humidification group and low VT group. Compared with high VT group,PaCO2 in high VT with airway humidification group was significantly decreased,Ppeak raised obviously,and no difference in pH value,PaO2,Raw and pulmonary compliance was found. Compared with low VT with airway humidification group,no difference in blood gas analysis(PaCO2,mmHg,1 mmHg=0.133 kPa)was found,but Ppeak(cmH2O,1 cmH2O=0.098 kPa),Raw(cmH2O),and lung compliance(mL/cmH2O)were increased significantly in high VT with airway humidification group(PaCO2 at 2 hours:27.96±4.64 vs. 36.08±2.11,4 hours:28.62±2.93 vs. 34.55±5.50, 6 hours:29.33±2.14 vs. 35.01±5.53;Ppeak at 0 hour:14.34±1.97 vs. 8.84±1.32,2 hours:17.33±0.52 vs. 11.17±2.14,4 hours:17.83±0.98 vs. 12.67±2.06,6 hours:18.67±1.22 vs. 13.50±2.16;Raw at 0 hour:37.36±5.14 vs. 27.05±2.93,2 hours:43.94±6.58 vs. 31.95±3.56,4 hours:48.04±6.07 vs. 35.24±3.50, 6 hours:50.33±6.34 vs. 36.66±3.64;pulmonary compliance at 6 hours:2.28±0.18 vs. 1.86±0.37,all P<0.05). The lung W/D ratio in high VT group was significantly higher than that of the low VT group(6.17±2.14 vs. 3.50±1.52, P<0.05). W/D in high VT with airway humidification group was higher than that of low VT with airway humidification group but without statistically significant difference(5.17±2.14 vs. 3.00±1.10,P>0.05). Microscopic observation showed that cilia were partially detached,adhered and sparse in low VT group,while cilia in high VT group showed serious detachment and lodging. Remaining cilia were sparse,with lodging,and cellular structure was damaged. Lung tissue pathological injury score in the high VT group was significantly higher than that of low VT group(6.17±2.14 vs. 3.50±1.52,P<0.05). Cilia density and cellularity were normal in low VT with airway humidification group,and no difference in lung tissue pathological injury score was found compared with low VT group(3.00±1.10 vs. 3.50±1.52, P>0.05). Cilia were severely detached,adhered and lodging,and cellularity were not obvious in high VT with airway humidification group,and lung tissue pathological injury score was elevated significantly than that of the low VT with airway humidification group but without statistically significant difference(5.17±2.14 vs. 3.00±1.10,P>0.05). TNF-α and IL-8 concentrations showed no change in plasma and BALF in all groups during ventilation,and no significant difference was found among the groups. Conclusions Airway humidification can alleviate pathological lung injury,damage of cilia and cellular structure in trachea caused by mechanical ventilation with low and high VT. High VT with humidification can result in serious pulmonary edema.

Objective To evaluate the clinical results of indirect reduction and fixation with the self-manufactured external fixator as a viable alternative in the surgical treatment of intraarticular calcaneal fractures.Methods From May 2006 to May 2009,a total of 30 patients undergone surgical treatment of intraarticular calcaneal fractures were analyzed,including 20 males and 10 females with an average age of 36 years (range,15-53).According to Sanders classification based on the computed tomography scan of intraarticular calcaneal fractures,16 patients were classified as type-Ⅲ,and 14 type-Ⅳ in this series.All fractures were treated first with the external fixator as indirect reduction and fixation device on the whole,which can enlarge the interspace of the subtalar joint significantly.Then,posterior articular facet of calcaneus was exposed and reduced through a small lateral incision.The calcaneal's length,breadth,thalamus height,maximum vertical displacement of the post-articular surface,and B(o)hler angle were measured preoperatively,3 days and 6 months after operation in X-ray film.Reduction results were evaluated by CT scan according to the standard of Buckley.Results The average follow-up time of all patients was 29 months (range,4-45).Lateral and axial roentgenograms showed satisfactory restoration of the calcaneal's anatomical structure.There were significant differences between preoperative values and those 3 days or 6 months postoperatively.There were no significant differences between values 3 days postoperatively and those 6 months postoperatively.The reduction results of posterior articular facet were evaluated by CT scan.Twenty-seven patients obtained anatomical reduction,3 patients obtained uneven articular facet within 2 mm.Conclusion This selfmanufactured external fixator is a vialbe alternative in the treatment of intraarticular calcaneal fractures,which has advantages of minimal invasion,practicality and less complications.