BACKGROUND:Calcium sulfate has good biocompatibility and biodegradability, which is a safe and effective bone graft substitute.
OBJECTIVE:To investigate the osteogenesis ability of calcium sulfate combined with bone marrow mesenchymal stem cells.
METHODS:After L4/5 posterior lumbar discectomy, 36 rabbits were randomized into three groups:rabbits in autologous bone group were implanted with autologous iliac bone via the intervertebral space;animals in al ogenic bone group were implanted with decalcified bovine bone;rabbits in tissue-engineered bone group were implanted with calcium sulfate combined with bone marrow mesenchymal stem cells. Bone formation and molding were observed by gross observation, anteroposterior and lateral X-ray, histology and biomechanics at 4, 8 and 16 weeks. Cal us specimens were employed for histological observation of interbody fusion. Biomechanical analysis of spinal fusion site was conducted at 16 weeks.
RESULTS AND CONCLUSION:Sixteen weeks later, interbody fusion was complete in the autologous bone group, the trabecular bone bridged continuously and a large amount of woven bone was merged into pieces;in the al ogenic bone group, incomplete bony fusion was found between the intervertebral space, most of cartilage tissues differentiated into bone, but fibrous tissue was also ful of the central part;in the tissue-engineered bone group, interbody fusion was complete, and a large amount of woven bone was fused into pieces, while the artificial bone was absorbed and ossified with smal residual. Failure strength and stiffness in the autologous bone and tissue-engineered bone groups were superior to those in the al ogenic bone group. These findings indicate that the calcium sulfate/bone marrow mesenchymal stem cells tissue-engineered bone has excellent osteogenic and osteoinductive capacity that can exert a good function of promoting spinal interbody fusion.

Acute promyelocytic leukemia(APL)is identified as the M3 subtype of acute myelocytic leukemia(AML). One of its clinical features is severe coagulation dysfunction.Most patients could develop disseminated intravascular coagulation(DIC)resulting vital viscera hemorrhage, which is one of the significant causes of early death in APL patients.Early diagnosis and inductive therapy, dynamic monitoring patients′ coagulation index and correcting the abnormal coagulation could prevent the occurrence or progress of DIC, and then improving the early-term survival rate of APL patients.At present, APL is mainly treated from the treatment of primary disease, anticoagulant antifibrinolytic therapy, and alternative therapy.

Objective To evaluate effects of camptothecin on the autophagy of HaCaT cells.Methods Some cultured HaCaT cells were divided into several groups to be treated with camptothecin at concentrations of 5,10,25,50,100 and 200 nmol/L,and 0.1％ dimethyl sulfoxide (DMSO) (control group),respectively.Cell counting kit-8 (CCK-8) assay was conducted to estimate the proliferative activity of HaCaT cells after 24-and 48-hour treatment,flow cytometry to evaluate cell apoptosis after 24-hour treatment,and Western blot analysis to measure the expression of autophagy-related proteins microtubuleassociated protein 1 light chain 3 (LC3) and p62.Some HaCaT cells were divided into 2 groups to be treated with 10 nmol/L camptothecin and 0.1％ DMSO for 24 hours,respectively.Then,indirect immunofluorescence assay (IFA) was performed to determine the LC3 expression.Results Camptothecin at low concentrations of 5 and 10 nmol/L had no significant effects on the proliferation and apoptosis of HaCaT cells.Compared with the control group,the cellular proliferative rates were significantly inhibited by (31.23 ± 1.00)％,(54.21 ± 8.10)％ and (66.75 ± 10.70)％ in the 50-,100-and 200-nmol/L camptothecin groups after 24-hour treatment respectively,and by (25.81 ± 5.99)％,(44.35 ± 5.32)％,(65.81 ± 8.28)％ and (73.23 ± 9.59)％ in the 25-,50-,100-and 200-nmol/L camptothecin groups after 48-hour treatment respectively (all P ＜ 0.001).After 24-hour treatment,the apoptosis rates were significantly higher in the 50-,100-and 200-nnol/L camptothecin groups (14.46％ ± 2.38％,19.15％ ± 1.59％,29.88％ ± 1.37％,respectively) than in the control group (3.80％ ± 0.13％,all P ＜ 0.001).After 24-hour treatment with 5 and 10 nmol/L camptothecin,the protein expression of LC3 Ⅱ was significantly up-regulated,while p62 protein expression was significantly down-regulated:IFA showed that the percentage of autophagosome-positive cells was significantly higher in the 10-nmol/L camptothecin group than in the control group after 24-hour treatment (36.67％ ± 4.55％ vs.6.23％ ± 0.92％,t =6.546,P =0.003).Conclusions Camptothecin at low concentrations of 5 and 10 nmol/L can induce autophagy of HaCaT cells,but has no obvious effects on cell proliferation and apoptosis.Camptothecin at concentrations of 50,100 and 200 nmol/L can inhibit cell proliferation,promote cell apoptosis,and decrease autophagy levels.

We describe an interactive viewer for the All-Species Living Tree (LVTree). The viewer incorporates treeing and lineage information from the ARB-SILVA website. It allows collapsing the tree branches at different taxonomic ranks and expanding the collapsed branches as well, keeping the overall topology of the tree unchanged. It also enables the user to observe the consequence of trial lineage modifications by re-collapsing the tree. The system reports taxon statistics at all ranks automatically after each collapsing and re-collapsing. These features greatly facilitate the compar-ison of the 16S rRNA sequence phylogeny with prokaryotic taxonomy in a taxon by taxon manner. In view of the fact that the present prokaryotic systematics is largely based on 16S rRNA sequence analysis, the current viewer may help reveal discrepancies between phylogeny and taxonomy. As an application, we show that in the latest release of LVTree, based on 11,939 rRNA sequences, as few as 24 lineage modifications are enough to bring all but two phyla (Proteobacteria and Firmicutes) to monophyletic clusters.

Objective: To investigate the characteristics of acoustic impedance test and its diagnostic role for occupational noise induced deafness, in order to provide an objective basis for the differential diagnosis of occupational noise induced deafness. Methods: A retrospective study was conducted to investigate the cases on the diagnosis of occupational noise-induced deafness in Guangdong province hospital for occupational disease prevention and treatment from January 2016 to January 2017. A total of 198 cases (396 ears) were divided into occupation disease group and non occupation disease group based on the diagnostic criteria of occupational noise deafness in 2014 edition, acoustic conductivity test results of two groups were compared including tympanograms types, external auditory canal volume, tympanic pressure, static compliance and slope. Results: In the occupational disease group, 204 ears were found to have 187 ears (91.67%) of type A, which were significantly higher than those in the non occupational disease group 143/192 (74.48%) , the difference was statistically significant (χ²=21.038, P<0.01). Detection of Ad or As type, occupation disease group in other type were 16/204 (7.84%) , 3/204 (1.47%) , were lower than Ad or As type of occupation disease group (15.63%) , other type (9.38%) , the differences were statistically significant[ (χ²=5.834, P<0.05) , (χ²=12.306, P<0.01) ]. Occupation disease group canal volume average (1.68±0.39) ml higher than that of non occupation disease group (1.57±0.47) ml, the difference was statistically significant (t=2.756, P<0.01) ; occupation disease group mean static compliance (1.06±0.82) ml higher than that of non occupation disease group (0.89±0.64) ml. The difference was statistically singificant (t=2.59, P<0.01) . Conclusion We observed that acoustic impedance test had obvious auxiliary function in the differential diagnosis of occupational noise induced deafness, More than 90% of the confirmed cases showed an A-form tympanograms, it is one of the objective examination methods which can be used in the differential diagnosis of pseudo deafness.

Objective To investigate the value of Chlamydia trachomatis antigen in patients with tubal infertility and its effect on the prevention of infection. Methods 80 patients with tubal infertility who were admitted in our hospital from January 2016 to October 2017 were selected as the study group and 80 patients with tubal patency by hysterosalpingography in our hospital were selected as the control group. The study group underwent Chlamydia trachomatis antigen detection. The relevant indicators were observed and clinical diagnostic effect and the effect of infection prevention of patients was recorded in detail. Results The diagnostic results showed that there were 77 cases diagnosed in the study group, and the diagnostic coincidence rate was 96. 2％, of which the diagnostic coincidence rates of tubal blockage, genital tract infection, tubal effusion, acute salpingitis were 100. 0％, 96. 1％, 96. 6％, 93. 8％. The positive rate of Chlamydia trachomatis antigen in the study group was 82. 5％ (66/80). The positive rate of Chlamydia trachomatis antigen in the control group was 15. 0％ (12/80). The positive rate of the study group was significantly higher than that of the control group (P＜0. 05). The test results showed that the incidence of abortion, preterm birth, dead fetus, stillbirth, ectopic pregnancy and other symptoms was significantly higher in the positive patients' study group. In the aspect of infection prevention, the number of patients infected at each time period in the study group gradually reduced with the growth of time and the increase of number of infected people. Conclusion The detection of Chlamydia trachomatis antigen in patients with tubal infertility can achieve high coincidence rate of clinical diagnosis, earlier detection of tubal disease in patients and timely treatment measures, reduce the incidence of postoperative infection in patients, which should be further applied in clinical.

Objective:To provide clinical evidence-based prognosis for patients with postpartum HEELP syndrome complicated with acute renal failure. Methods:The clinical question was proposed. Some database was searched to assess the evidence. Results:A total of 4 studies were included,however,the quality of the studies was low. The results of the studies was suitable for our patients. The death of patients with postpartum HEELP syndrome complicated with acute renal failure was attributed to multiple organ failure. The risk factors for poor prognosis was primipara, delivery after twin pregnancy, absence of typical symptoms, rapid clinical progression, high blood pressure, epigastric pain. Early assessment and diagnosis for maternal and infant, positive in intervention and be caution for the DIC and multiple organ failure, using plasmapheresis can improve the prognosis. Conclusion:Patients with postpartum HEELP syndrome complicated with acute renal failure have an acute process and poor prognosis, especially for patients without typical symptoms.

Objective: Study of the results and the degree on occupational noise-induced deafness in-to the different high frequency hearing threshold weighted value, in order to provide theoretical basis for the re-vision of diagnostic criteria on occupational noise-induced deafness. Methods: A retrospective study was con-ducted to investigate the cases on the diagnosis of occupational noise-induced deafness in Guangdong province hospital for occupational disease prevention and treatment from January 2016 to January 2017. Based on the re-sults of the 3 hearing test for each test interval greater than 3 days in the hospital, the best threshold of each frequency was obtained, and based on the diagnostic criteria of occupational noise deafness in 2007 edition, Chi square test, t test and variance analysis were used to measure SPSS21.0 data, their differences are tested among the means of speech frequency and the high frequency weighted value into different age group, noise ex-posure group, and diagnostic classification between different dimensions. Results: 1. There were totally 168 cases in accordance with the study plan, male 154 cases, female 14 cases, the average age was 41.18 ±6.07 years old. 2. The diagnosis rate was increased into the weighted value of different high frequency than the mean value of pure speech frequency, the weighted 4 kHz frequency increased by 13.69% (χ²=9.880, P=0.002) , 6 kHz increased by 15.47% (χ²=9.985, P=0.002) and 4 kHz+6 kHz increased by15.47% (χ²=9.985, P=0.002) , the difference was statistically significant. The diagnostic rate of different high threshold had no obvious differ-ence between the genders. 3. The age groups were divided into less than or equal to 40years old group (A group) and 40-50 years old group (group B) , there were higher the diagnostic rate between high frequency weighted 4 kHz (A group χ²=3.380, P=0.050; B group χ²=4.054, P=0.032) , weighted 6 kHz (A group χ²=6.362, P=0.012; B group χ²=4.054, P=0.032) , high frequency weighted 4 kHz+6 kHz (A group χ²=6.362, P=0.012; B group χ²=4.054, P=0.032) than those of speech frequency average value in the same group on oc-cupational noise-induced deafness diagnosis rate, the difference was statistically significant. There was no sig-nificant difference between age groups (χ²=2.265, P=0.944) . 4. The better ear's mean value of pure speech fre-quency and the weighted values into different high frequency of working years on each group were compared, working years more than 10 years group was significantly higher than that of average thresholds of each frequen-cy band in 3-5 group (F=2.271, P=0.001) , 6-10 group (F=1.563, P=0.046) , the difference was statistically significant. The different high frequency weighted values were higher than those of the mean value of pure speech frequency, and the high frequency weighted 4 kHz+6 kHz had the highest frequency difference, with an average increase of 2.83 dB. 5. The diagnostic rate into weighted different high frequency was higher in the mild, moderate and severe grades than in the pure speech frequency. In the comparison of diagnosis for mild occupational noise-induced deafness, in addition to the weighted 3 kHz high frequency (χ²=3.117, P=0.077) had no significant difference, the weighted 4 kHz (χ²=10.835, P=0.001) , 6 kHz (χ²=9.985, P=0.002) , 3 kHz+4 kHz (χ²=6.315, P=0.012) , 3 kHz+6 kHz (χ²=6.315, P=0.012) , 4 kHz+6 kHz (χ²=9.985, P=0.002) , 3 kHz+4 kHz+6 kHz (χ²=7.667, P=0.002) were significantly higher than the diagnosis rate of the mean value of pure speech frequency. There was no significant difference between the two groups in the moderate and se-vere grades (P>0.05) . Conclusion Bring into different high frequency hearing threshold weighted value in-creases the diagnostic rate of occupational noise-induced deafness, the weighted 4 kHz, 6 kHz and 4 kHz+ 6 kHz high frequency value affects the result greatly, and the weighted 4 kHz+6 kHz high frequency hearing threshold value is maximum the effect on occupational noise-induced deafness diagnosis.

Objective:Patients with acute promyelocytic leukemia(APL)treated with arsenic trioxide(ATO)have significant coagulation disorders both in pretherapy and post-treatment, and severe cases can progress quickly to the disseminated intravascular coagulation(DIC). This article aims to investigate the risk factors of DIC and observe dynamically the effect of ATO on coagulation function in APL children.Methods:We collected basic characteristics, laboratory characteristics and coagulation index at various time points on admission of 28 children who were first diagnosed APL and treated with ATO at department of pediatric hematology in Shengjing Hospital from January 2012 to January 2018. We retrospectively analyzed the correlation between these factors and the occurrence of DIC, and the trend of dynamic changes in coagulation index in different treatment time.Results:Of the 28 patients, 24(85.7%)were diagnosed as DIC. No relation has been found between the occurrence of DIC and the age, gender, blood routine at initial diagnosis, fibrinogen level, lactate dehydrogenase level in APL children( P>0.05). Different indicators of plasma prothrombin time, fibrinogen at different time point showed significantly difference. In the high-risk group, PT was higher significantly before treatment[(17.58±2.99)s] than those of the 3-4 days after therapy[(15.3±1.86)s], 7-10 days after therapy[(13.81±1.18)s], 14-16 days after therapy[(14.13±0.62)s], 22-26 days after therapy[(14.2±1.59)s]( P=0.016). In the group which PLT was greater than or equal to 20×10 9/L, PT was higher significantly before treatment[(15.02±2.83)s]and the 3-4 days after therapy[(14.59±2.19)s] than those of the 7-10 days after therapy[(13.25±2.01)s], 14-16 days after therapy[(13.31±1.51)s], 22-26 days after therapy[(13.47±1.64)s]( P<0.01). Conclusion:No relation has been found between the occurrence of DIC and the age, gender, blood routine at initial diagnosis, fibrinogen level, lactate dehydrogenase level in APL children. The time points of significantly abnormal of coagulation function occurred in the newly diagnosed APL children were before the ATO induction treatment and the 3-4 days after therapy.

AIM:To explore the effects of CD38 on lysosome reformation and cholesterol efflux in macro-phages.METHODS:Bone marrow-derived macrophages from low-density lipoprotein(LDL)receptor knockout(LDLr-/-)mice were cultured as cell model.Live cell imaging system was applied to evaluate the effect of nicotinic acid adenine di-nucleotide phosphate(NAADP)on lysosome number.ELISA was conducted to measure NAADP level in macrophages.After the cells were treated with nicotinic acid(NA),RT-qPCR was conducted to detect CD38 mRNA expression,and Western blot was conducted to observe CD38 protein expression and phosphorylated transcription factor EB(TFEB)level.Laser scanning confocal microscopy was applied to evaluate the influence of CD38/NAADP signaling on lysosome number and cholesterol egression.RESULTS:NAADP remarkably increased lysosome number(P<0.05),and this effect was significantly inhibited by NAADP antagonist NED-19,Ca2+ chelator BAPTA,and calcineurin inhibitor CsA(P<0.05).CD38 markedly enhanced NAADP synthesis in macrophages(P<0.05).NAADP synthetic substrate NA prominently ele-vated the expression of CD38 mRNA and protein(P<0.05).NA significantly decreased the phosphorylated TFEB level;this effect was also attenuated by NED-19,BAPTA and CsA(P<0.05).Disrupting CD38/NAADP signaling pathway markedly inhibited NA-induced enhancement of lysosome number,lysosomal free cholesterol and cytosol cholesterol ester efflux in macrophages(P<0.05).NA-induced enhancement of lysosome number,lysosomal free cholesterol and cytosol cholesterol ester efflux abolished in LDLr/CD38 DKO macrophages(P<0.05),whereas these effects induced by NA were recovered after CD38 gene rescue.CONCLUSION:CD38 triggers lysosome reformation via TFEB and consequently pro-motes the efflux of lysosomal free cholesterol and cytosol cholesterol ester.