Objective: To investigate the etiology spectrum of hand foot and mouth disease (HFMD) and to analyze the molecular characteristics of Coxsackievirus A10 and A6 in Qingdao in 2014. Methods: Throat swabs of HFMD cases were tested for total enteroviruses (EVs), EV-A71, CV-A16, CV-A10 and CV-A6 by multiplex real time RT-PCR. Other EV serotypes were identified through the sequences of partial VP1 gene. The full-length of VP1 gene of CV-A10 and CV-A6 were amplified and sequenced. The sequences were phylogenetically analyzed using MEGA 5.0 software package. Results: A total of 1727 HFMD patients were detected in 2014 and 11serotypes of enteroviruses were identified. EV-A71(38.0%, 410/1078), CV-A16(28.8%, 311/1078), CV-A10(14.1%, 152/1078)and CV-A6(3.2%, 34/1078)were the most dominant pathogen in 2014 in Qingdao. Proportions of CV-A10 in enterovirus infected children varied dramatically in different ages(χ²=15.19, P=0.001), showing a downtrend with age. Proportion of CV-A10 in inpatients was much higher than that in outpatients(χ²=49.1, P <0.001), while 60% of those inpatients showed nervous system damage symptoms, with pathological reflex or disappearance of physiological reflex. Phylogenetic analysis of complete VP1 sequences showed that all CV-A10 strains identied in this study belonged to genotype C, 71.7% of which located in branch C5; all CV-A6 strains belonged to genotype D while 83.3% of which located in branch D5. Conclusions EV-A71, CV-A16, CV-A10 and CV-A6 were the prevalent pathogens of HFMD in Qingdao in 2014. CV-A10 was more frequently detected in lower age group with HFMD, which can cause damage to nervous system. Strains of branch C5 in genotype C were the dominant strains of CV-A10 circulated in Qingdao in 2014 while strains of branch D5 in genotype D were the major strains of CV-A6.

Objective To investigate the epidemic situation and its epidemic characteristics of malaria in Qingdao City from 2012 to 2017, so as to provide the evidence for the prevention and control of malaria in this city. Methods The data of cases and epidemiological characteristics of malaria in Qingdao City, 2012–2017 were collected from the China’s disease prevention and control system infectious disease report information management system and parasitic disease prevention and control information management system, and analyzed. Results A total of 103 cases of malaria were reported in Qingdao City from 2012 to 2017, and all of them were imported. Among them, 98 (95.15%) were imported from Africa and 5 (4.85%) from Asia. There were 92 laboratory confirmed cases (89.32%) and 11 (10.68%) clinically diagnosed cases. The highest incidence occurred in 2016 (0.28/105) . According to the classification of cases, there were 87 falciparum malaria cases (84.47%), 13 vivax malaria cases (12.62%), and 3 ovale malaria cases (2.91%) . These patients were mainly young men, and the male to female ratio was 19.6∶1. There were malaria case reports in each month, and the peak onset season was not obvious. Newly diagnosed and confirmed units were mainly municipal medical institutions; the median of onset-definite diagnosis was 4 days. There was one death case, but there were no recurrence, reburning, and imported secondary cases. Conclusion There were no local malaria cases in Qingdao City from 2012 to 2017. The prevention and control of imported malaria is the focus of the future work.

Fractures, Bone ; Humans ; Osteoarthritis, Knee ; Osteotomy ; Tibia/surgery* ; Tibial Fractures/surgery*

Humans ; Female ; Breast Neoplasms

OBJECTIVE: To explore the clinical application value of artificial intelligence image aided diagnosis platformbased on Faster R-CNN in identifying EMVI of rectal cancer.METHODS: In the multicenter retrospective study,500 patients with rectal cancer who underwent high-resolution MRI examination between July 2016 and February 2019 wereselected from seven hospitals in China. They were divided into 174 positive and 326 negative patients. Patients wererandomized to a training group(400 patients,including 133 positive and 267 negative) and a validation group(100 patients,including 41 positive and 59 negative) using a random number method. Using the Faster R-CNN to learn and train 20 430 high-resolution MRI images of thetraining group,an artificial intelligence image-aided diagnosis platform was established. The5107 high-resolution MRI images of thevalidation group were clinically validated.Receiver operating characteristic(ROC) curveand area under the curve(AUC) were used tocompare the diagnostic results of the artificialintelligence image-aided diagnosis platform andthe senior image expert.RESULTS: The accuracy,sensitivity,specificity,positive predictive valueand negative predictive value of EMVI forartificial intelligence image-aided diagnosis platform were 93.4%, 97.3%, 89.5%, 0.90 and 0.97,respectively. The area under the receiver operating characteristiccurve(AUC) was 0.98. The time required to automatically recognize a single image was 0.2 seconds,which had clearadvantages compared to radiologists(estimated to be about 10 seconds).CONCLUSION: The artificial intelligence image-assisted diagnosis platform based on Faster R-CNN has high efficiency and feasibility for identifying rectal cancerEMVI,and can assist imaging diagnosis.

The pathogenesis of hepatocellular carcinoma is complex and has not been fully clarified. KLF4, also known as gut-enriched Kruppel-like factor (GKLF), plays the dual regulatory role of tumor suppressor and tumor promoter. KLF4 plays an anti-cancer role in liver cancer. This article introduces the biological function and regulatory mechanism of KLF4 in the progression of liver cancer, and research on the mechanism of action of KLF4 in liver cancer is expected to provide new ideas for targeted therapy and prognosis monitoring of liver cancer.

Objective: To analyze the molecular epidemiology of norovirus (NoV) genotype GⅡ.15 in Qingdao City. Methods: One thousand four hundred and twelve stool samples were collected from suspected NoV infected patients and detected by real-time polymerase chain reaction (PCR). Open reading frame (ORF)1-ORF2 and VP1 gene were amplified by reverse transcription (RT)-PCR and sequenced for genotyping, evolutionary analysis and homology modeling. Results: Seven cases of GⅡ.15 type were detected including four sporadic cases and one outbreak.The VP1 gene was highly homologous and had little variation compared with early strain J23/US/1999. The differences of amino acids between strains in Qingdao City were mainly asparagine/asparticacid(N/D)300 and proline/serine(P/S)302.Homology modeling suggested that VP1 of GⅡ.15 strain was composed of S domain and P domain (P1 subdomain included 224-276 and 431-555, P2 subdomain included 277-430). S domain contained eight anti-parallel β-sandwiches and two α-helixes, and P1 subdomain contained one α-helix and seven β-strands, and the P2 subdomain folded into a compact barrel-like structure consisting of six β-strands.Argnine (R)-glycine (G)-valine (V)-motif (289-291) and three specific loci including glutarnine (Q)313, asparagine (N)349 and Q389 were located in the P2 subdomain, with NGR-motif (265-267) located at 22nd upstream of RGV-motif.Site I (SNR-alanine(A)- histidine(H)357-361), Site Ⅱ (D388) and Site Ⅲ (G454, G455) were the main characteristic sites of histo-blood group antigens (HBGA) binding interface, which may be similar to the binding pattern of GⅡ.4 type VA387 and HBGA. Conclusion Although GⅡ.15 type NoV evolves very slowly, it may still have the risk to become an epidemic strain, which needs to be monitored and further studied.

Objective: To analyze the molecular epidemiology of Norovirus(NoV) infection among sporadic hospitalized adults with diarrhea in Qingdao, 2015. Methods: Four hundred and nine stool samples were collected from hospitalized adults with diarrhea and detected by Real-time RT-PCR. For genotyping, ORF1 and ORF2 were partially amplified by RT-PCR and sequenced. Results: 18.1%(74/409) of stool samples were positive for NoV genogroup I(GI) (10/74) and genogroup II(G II) (64/74). Fifty-three positive samples of GII were sequenced and divided into 4 genotypes, including GII.Pe-GII.4(26/53), GII.P17-GII.17(19/53), GII.P12-GII.3(7/53) and GII.P16-GII.13(1/53). All GII.4 strains were the variants of GII.4-Sydney-2012. From May to Aug in 2015, GII.4 only accounted for 5.3%(1/19) and GII.17 were the major epidemic strains(68.4%, 13/19). But from Sept to Dec in 2015, GII.17 decreased substantially to 10.9%(6/55) and GII.4 became the most predominated strains (45.5%, 25/55). Conclusions NoV is an important pathogen responsible for viral diarrhea among adults in Qingdao. GII.4-Sydney-2012 and the newly variant GII.P17-GII.17 were the predominant epidemic strains in Qingdao, 2015.

OBJECTIVETo investigate the effects of human umbilical cord blood-derived mesenchymal stem cells(HUCMSC) on the leukemic cell line HL-60 and acute lymphoblastic leukemia cell line Jurkat as well as the role of CXCL12/CXCR4.

METHODSHL-60 cells and Jurkat cells were co-cultured with human umbilical cord blood mesenchymal stem cell (HUCMSC), and the model was treated with G-CSF, AMD3100 and their combination. The cell viability and cell cycle were measured by Cell Counting Kit-8 (CCK-8), the apoptosis and the cell-cycle analysis were assessed by flow cytometry with the Annexin V/PI double staining. The expression of surface CXCR4 protein and total CXCR4 protein of leukemic cells were detected by flow cytometry and Western blot respectively.

RESULTSHUCMSC could decrease the viability of HL-60 cells and Jurkat cells, as well as the percentage of apoptotic cells, they could also increase the number of G/Gcells, while G-CSF and AMD3100 could reduce the proliferation of HL-60 cells and Jurkat cells in HUCMSC co-culture model, destructed the anti-apoptotic effect of HUCMSC on HL-60 cells and Jurkat cells, and the combination of 2 drugs resulted in a synergistic effect. The G-CSF could reduce the expression of surface CXCR4 protein and total CXCR4 protein in leukemic cells, while AMD3100 could only decrease the expression of surface CXCR4 protein of leukemia cell membrane, having no effect on the expression of CXCR4 protein in cytoplasm.

CONCLUSIONHuman umbilical cord blood mesenchymal stem cells can inhibit the proliferation and apoptosis of acute leukemia cells and increase the number of G/Gphase cells in leukemic cells. The AMD3100 can decrease the expression of surface CXCR4 protein in leukemia cells, G-CSF can decrease expression of total CXCR4 protein as well as membrane CXCR4 protein. Both of them can block the CXCL12/CXCR4 signal axis, weakening the relationship between leukemia cells and microenvironment. And on the basic of HUCMSC influenced leukemia cells' growth and proliferation, the cell viability will be weakened, its apoptosis will be promoted, and the percentage of G/Gphase cells in leukemia cells will be decreased.

This study was aimed to investigate the effects of the DNA methylation inhibitor 5-aza-2'-deoxycytidine (5-Aza-CdR) and histone deacetylase inhibitor trichostatin A (TSA) on DLC-1 gene transcription regulation and molecular biological behaviours in the human multiple myeloma RPMI-8226 cells. The cells were treated respectively with 5-Aza-CdR and TSA alone, or the both combination; the cell proliferation and apoptosis, DLC-1 expression, the protein expression of Ras homolog family member A (RhoA) and Ras-related C3 botulinum toxin substrate 1 (Rac1) were examined by CCK-8 method, RT-PCR and ELISA, respectively. The results showed that the 5-Aza-CdR and TSA had cell growth inhibitory and apoptosis-inducing effects in dose-dependent manner (P < 0.05). Compared with a single drug (5-Aza-CdR or TSA alone), the effects were significantly enhanced after treatment with the combination of 5-Aza-CdR and TSA (P < 0.05). DLC-1 was weakly expressed in the control group; the treatment with 5-Aza-CdR alone enhanced its re-expression dose-dependently (P < 0.05). Compared with 5-Aza-CdR alone, 5-Aza-CdR plus TSA enhanced DLC-1 re-expression significantly.Compared with the control, 5-Aza-CdR and TSA significantly decreased RhoA and Rac1 protein expression (P < 0.05). It is concluded that 5-Aza-CdR and TSA can effectively reverse DLC-1 expression of RPMI-8226 cells; TSA has a synergistic effect on its re-expression. 5-Aza-CdR and TSA have significant cell growth inhibitory and apoptosis-inducing effects on RPMI-8226 cells. These effects may be related to the inhibition of Rho/Rho kinase signalling pathway.
Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Azacitidine ; administration & dosage ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; GTPase-Activating Proteins ; metabolism ; Gene Expression ; drug effects ; Humans ; Hydroxamic Acids ; administration & dosage ; pharmacology ; Multiple Myeloma ; genetics ; pathology ; Tumor Suppressor Proteins ; metabolism