Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.

OBJECTIVE: To detect the differential expressions of miR-451, ABCB1 and ABCC2 in drug-sensitive leukemia cell line K562 and drug-resistant cell line K562/A02, and explore the regulatory relationship between miR-451 and the expressions of ABCB1 and ABCC2 , and the mechanism of miR-451 involved in drug resistance in leukemia. METHODS: CCK-8 assay was used to detect the drug resistance of K562/A02 and K562 cells. Quantitative Real-time PCR (qRT-PCR) was used to verify the differential expressions of miR-451 in K562 and K562/A02 cells. MiR-451 mimic and negative control (miR-NC), miR-451 inhibitor and negative control (miR-inNC) were transfected into K562 and K562/A02 cells respectively, then qRT-PCR and Western blot were used to detect the expression levels of mRNA and protein of ABCB1 and ABCC2 in K562 and K562/A02 cells and the transfected groups. RESULTS: The drug resistance of K562/A02 cells to adriamycin was 177 times higher than that of its parent cell line K562. Compared with K562 cells, the expression of miR-451 in K562/A02 cells was significantly higher (P <0.001), and the mRNA and protein expression levels of ABCB1 and ABCC2 in K562/A02 cells were significantly higher than those in K562 cells (P <0.001). After transfected with miR-451 inhibitor, the expression of miR-451 was significantly down-regulated in K562/A02 cells (P <0.001), the sensitivity to chemotherapy drugs was significantly enhanced (P <0.05), and the mRNA and protein expressions of ABCB1 and ABCC2 were significantly decreased (P <0.01). After transfected with miR-451 mimic, the expression of miR-451 was significantly upregulated in K562 cells (P <0.001), and the mRNA and protein expressions of ABCB1 and ABCC2 were significantly increased (P <0.01). CONCLUSION There are significant differences in the expressions of miR-451, ABCB1 and ABCC2 between the drug-sensitive leukemia cell line K562 and drug-resistant cell line K562/A02, which suggests that miR-451 may affect the drug resistance of leukemia cells by regulating the expression of ABCB1 and ABCC2.
Humans ; K562 Cells ; Drug Resistance, Neoplasm/genetics* ; Drug Resistance, Multiple/genetics* ; Doxorubicin/pharmacology* ; MicroRNAs/genetics* ; Leukemia/genetics* ; RNA, Messenger

OBJECTIVE: To analyze the characteristics of gene mutation and overexpression in newly diagnosed multiple myeloma (NDMM) patients. METHODS: Bone marrow cells from 208 NDMM patients were collected and analyzed. The gene mutation of 28 genes and overexpression of 6 genes was detected by DNA sequencing. Chromosome structure abnormalities were detected by fluorescence in situ hybridization (FISH). RESULTS: Gene mutations were detected in 61 (29.33%) NDMM patients. Some mutations occurred in 5 or more cases, such as NRAS, PRDM1, FAM46C, MYC, CCND1, LTB, DIS3, KRAS, and CRBN. Overexpression of six genes (CCND1, CCND3, BCL-2, CCND2, FGFR3, and MYC) were detected in 83 (39.9%) patients, and cell cycle regulation gene was the most common. Single nucleotide polymorphisms (SNP) changes were detected in 169 (81.25%) patients, the TP53 P72R gene SNP (70.17%) was the most common. Abnormality in chromosome structure was correlated to gene overexpression. Compared to the patients with normal chromosome structure, patients with 14q32 deletion showed higher proportion of CCND1 overexpression. Similarly, patients with 13q14 deletion showed higher proportion of FGFR3 overexpression, whereas patients with 1q21 amplification showed higher proportion of CCND2, BCL-2 and FGFR3 overexpression. CONCLUSION There are multiple gene mutations and overexpression in NDMM. However, there is no dominated single mutation or overexpression of genes. The most common gene mutations are those in the RAS/MAPK pathway and the genes of cyclin family CCND are overexpression.
Chromosome Aberrations ; Humans ; In Situ Hybridization, Fluorescence ; Multiple Myeloma/genetics* ; Mutation

This research explored the synergistic effects and the potential mechanisms of RCE-4 and various nonsteroidal anti-inflammatory drugs (NSAIDs) on the proliferation of cervical cancer Ca Ski cells. The MTT assay and CalcuSyn V2.0 software were used to detect cell proliferation and calculate the combination index (CI); the expression levels of various proteins were analyzed using Western blot assay; mitochondrial membrane potential (MMP) was assessed using JC-1 staining; acridine orange/ethidium bromide (AO/EB) double-fluorescence staining was used to detect the apoptosis of Ca Ski cells; a co-immunoprecipitation (Co-IP) assay was used to analyze the relative content of Bcl-2-Beclin 1 complex in Ca Ski cells. The results demonstrate that the combination of RCE-4 and NSAIDs increases the inhibition of Ca Ski cells compared to the single-RCE-4 group, and celecoxib provided the best synergistic effect among the four NSAIDs tested, with a CI of 0.32. The combination of RCE-4 and celecoxib significantly down-regulated the expression of cyclooxygenase-2 (COX-2) and nuclear transcription factor-κB (NF-κB), and promoted the expression of non-steroidal anti-inflammatory drugs activity gene-1 (NAG-1). In addition, autophagy induced by RCE-4 was markedly inhibited in combination with celecoxib, which was associated with down-regulation of the expression of microtubule-associated protein 3 (LC3)-II, Beclin 1, p62 and autophagy-related gene (ATG) 3/4B/5/7/14. RCE-4-induced apoptosis was significantly enhanced by altering the depolarization of mitochondrial membrane potential and the expression of B cell lymphoma-2 (Bcl-2), B cell lymphoma-xl (Bcl-xl), Bcl-2 associated X protein (Bax), Bcl-2/Bcl-xl-associated death promoter (Bad) and cleaved cysteinyl aspartate specific proteinase (cleaved-caspase) 3/7/9. Furthermore, the formation of the Bcl-2-Beclin 1 complex was significantly inhibited in Ca Ski cells treated with RCE-4 in combination with celecoxib. Taken together, this research shows that the combination of RCE-4 and celecoxib has a significant synergistic effect on the proliferation of Ca Ski cells by promoting apoptosis, inhibiting autophagy and disturbing the formation of the Bcl-2-Beclin 1 complex, which may be a novel strategy to increase the sensitivity of anti-cervical cancer drugs.

Objective To study the role of partial pressure of oxygen in the treatment of pulmonary hypertension induced by meconium aspiration syndrome. Methods Retrospective analysis was performed on 58 patients with meconium aspirate and pulmonary hypertension from January 1,2007 to December 31,2016, 28 patients in the death group,and 30 patients in the survival group. Blood gas analysis about two groups of children with mechanical ventilation were collected at four different time points:before mechanical ventilation ( 0 h) ,12 hours after mechanical ventilation,24 hours after mechanical ventilation and 72 hours after mechan-ical ventilation. The 80 mmHg(1 mmHg=0. 133 kPa) in blood gas analysis of the partial pressure of oxygen was used as the boundary point to study the effects about different blood oxygen partial pressures on the length of mechanical ventilation time in the survival group. Results Blood gas analysis showed that there were no significant differences in pH,partial pressure of oxygen,HCO3-,and lactic acid before mechanical ventilation in the two groups,but the pH values were significantly higher in the survival group after 12 h, 24 h,and 72 h of mechanical ventilation(t values:-2. 07,-3. 48,-7. 11;P <0. 05). Partial pressure of oxygen were higher in the surviving group at 12 h,24 h,and 72 h than those in the death group(t values-2. 87,-4. 88,-11. 29; P<0. 05). The HCO3-values of the survival group at 12 h,24 h and 72 h were higher than the death group, and the differences were statistically significant ( t values: -3. 90, -5. 60,-5. 76,P<0. 001). The lactic acid of survival group was significantly lower than the death group at 72 h after ventilation (t=5. 12,P<0. 001). Compared with the death group,partial pressure of carbon dioxide in 24 h,72 h decreased significantly in the surviving group(t values:5. 67,7. 60;P<0. 05). In the surviving group,the partial pressure of oxygen was maintained above 80 mmHg in 18 patients and the mechanical venti-lation time was (7. 17 ± 1. 95) d. The partial pressure of oxygen was below 80 mmHg in 12 patients of the surviving group and the mechanical ventilation time was (8. 67 ± 2. 50)d. The difference was statistically sig-nificant(t =12. 02,P <0. 001). Conclusion In the treatment of newborn with pulmonary hypertension caused by meconium aspiration,maintaining a higher pH value and a partial pressure of oxygen higher than 80 mmHg may be beneficial to the recovery of children with pulmonary hypertension.

Objective To investigate the impact of fasting blood glucose on the prognosis of non-diabetic patients undergoing primary percutaneous coronary intervention (PCI) for acute ST-segment elevation myocardial infarction (STEMI). Methods We retrospectively recruited consecutive patients who underwent primary PCI in our hospital from February, 2003 to March, 2015. Patients with prior history of diabetes mellitus before the index hospitalization and those with newly diagnosed diabetes mellitus during the index hospitalization were excluded. The clinical and angiographic features, medical and interventional treatment, and 30-day outcomes were compared between patients with elevated fasting blood glucose (FBG) (>5.4 mmol/L) and those with normal FBG (≤5.4 mmol/L). Results A total of 721 patients were recruited with an age of(61.2 ± 12.8)years, of whom 601 (83.4 %) were male. As compared with patients with normal FBG,those with elevated FBG were more likely to be female(20.1 % vs.13.5 %, P=0.017),had faster heart rate on admission[(82.9 ± 17.2)bpm vs.(79.4 ± 16.7)bpm,P=0.006]and more use of intra-aortic balloon pump(3.8 % vs.1.3 %,P=0.034),and had higher rates of 30 day all-cause mortality(3.5 % vs.0.5 %,P=0.004),cardiac mortality(2.9 % vs.0.5 %,P=0.012)and heart failure(18.1 % vs. 7.4 %, P< 0.001). After adjusting baseline characteristics, FBG > 5.4 mmol/L was one of the independent predictors of 30-day all-cause mortality(HR 6.030,95 % CI 1.235-29.447,P=0.026).Other independent predictors of 30-day all-cause mortality included age(HR 1.059,95 % CI 1.002-1.120,P=0.044),heart rate on admission(HR 1.036,95 % CI 1.003-1.070,P=0.034),left descending artery as the culprit vessel(HR 6.427,95 % CI 1.389-29.728,P=0.017),and use of angiotensin converting enzyme inhibitor/angiotensin receptor blocker(HR 0.154,95 % CI 0.051-0.461,P=0.001).Conclusions In non-diabetic patients undergoing primary PCI for STEMI, elevated FBG was one of the independent predictors of 30-day all-cause mortality.

OBJECTIVEWe aimed to evaluate the combined effect of a family history of cardiovascular disease (CVD) and high serum C-reactive protein (CRP) on the stroke incidence in an Inner Mongolian population in China.

METHODSA prospective cohort study was conducted from June 2002 to July 2012, with 2,544 participants aged 20 years and over from Inner Mongolia, China. We categorized participants into four groups based on the family history of CVD and CRP levels.

RESULTSWe adjusted for age; sex; smoking; drinking; hypertension; body mass index; waist circumference; and blood glucose, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol levels. Compared with the group with no family history of CVD/low CRP levels, the group with family history of CVD/high CRP levels had a hazard ratio (HR) of 1.78 [95% confidence interval (CI), 1.03-3.07; P = 0.039] of stroke, and an HR of 2.14 (95% CI, 1.09-4.20; P = 0.027) of ischemic stroke. The HRs of hemorrhagic stroke for the other three groups were not statistically significant (all P > 0.05).

CONCLUSIONParticipants with both a family history of CVD and high CRP levels had the highest stroke incidence, suggesting that high CRP levels may increase stroke risk, especially of ischemic stroke, among individuals with a family history of CVD.

Asian Continental Ancestry Group ; C-Reactive Protein ; metabolism ; Cardiovascular Diseases ; epidemiology ; genetics ; China ; Genetic Predisposition to Disease ; Humans ; Prospective Studies ; Risk Factors ; Stroke ; epidemiology

Anaprazole is a proton pump inhibitor clinically used for curing peptic ulcer. A rapid, sensitive and convenient LC-MS/MS method was first established for the determination of anaprazole in human plasma. d₃, ¹³C-anaprazole was used as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all components were separated on an Extend C₁₈ column (100 mm×4.6 mm, 3.5 μm). The assay was linear over the concentration range of 5.00-3 000 ng·mL^-1 (r² > 0.995). The method was successfully applied to a pharmacokinetic study of 40 mg anaprazole enteric-coated tablets in 14 Chinese healthy volunteers under fasting or high fat diet conditions. C_max was (1 020±435) ng·mL^-1 and AUC_0-t was (2 370±754) h·ng·mL^-1 under fasting condition. And C_max was (538±395) ng·mL^-1 and AUC_0-t was (1 610±650) h·ng·mL^-1 under high fat diet condition. The plasma results suggest that the exposure of anaprazole is reduced by the high fat diet.

Objective To explore the effect of salmon calcitonin on osteoporosis induced by spinal cord injury. Methods 100 patients with osteoporosis induced by spinal cord injury from September 2011 to September 2014 in our department were included. They were randomly divided into control group (n=50) and observation group (n=50). The control group received vitamin D3 only, while the observation group received vitamin D3 combined with salmon calcitonin on the basis of rehabilitation physiotherapy, for 6 months. Visual Analogue Scale (VAS) of pain was evaluated in different periods. The bone mineral density (BMD) of lumbar spine and femoral neck, the parathyroid hormone (PTH), bone gla protein (BGP) and 1,25- dihydroxy vitamin D3 (1,25-(OH)2D3) were tested and recorded. Results The VAS score was lower in the observation group than in the control group 1, 2, 3 and 6 months after treatment (P<0.001). The BMD of lumbar spine and femoral neck was significantly higher, the PTH and BGP were significantly lower and the 1,25-(OH)2D3 was significantly higher in the observation group than in the control group after treatment (P<0.001). Conclusion Combination of salmon calcitonin can effectively reduce the bone pain and improve the BMD in patients with osteoporosis induced by spinal cord injury.

OBJECTIVETo evaluate the effectiveness of locking plate external fixator in treating middle and distal tibial fractures.

METHODSFrom January 2010 to January 2013,18 patients suffered from middle and distal tibial fractures were treated by locking plate external fixator,including 11 males and 7 females, with an average age of 53.5 (ranged from 13 to 80) years old,the course of disease ranged from 2 h to 3 d. According to AO classification, 4 cases were type A,11 cases were type B and 3 were type C. Among them,6 patients were open fracture, including 2 cases with type I, 3 cases with type II and 1 case with type III, according to Gustilo classification), 12 patients were close fracture. Operation time, postoperative complications were observed, and Johner-Wruhs scoring were used to evaluate clinical outcomes.

RESULTSAll patients were followed up from 6 to 15 (meaned 11) months. Two cases occurred skin necrosis (1 case occurred bone exposure), 2 cases occurred delayed union (all were open fracture), and 1 case occurred nail infection. No screw loosening or broken occurred. According to Johner-Wruhs scoring, 10 cases obtained excellent result,6 cases good,and 2 cases fine.

CONCLUSIONLocking plate external fixator for the treatment of middle and distal tibial fractures, which has advantages of lessen damage, shorter operative time, less complications and rapid functional recovery, is one of good choice.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Plates ; utilization ; External Fixators ; utilization ; Female ; Fracture Fixation ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult