Objective To construct and identify the induced pluripotent stem cells(iPSCs)derived from peripheral blood mononuclear cells(PBMCs)of a patient with facioscapulohumeral muscular dystrophy(FSHD),initially explore their differentiation ability into skeletal muscle cells,and evaluate the feasibility of using this cell model for disease mechanism research.Methods The PBMCs from one FSHD patient were collected and transfected with Sendai virus containing four reprogramming transcription factors,including OCT4,SOX2,KLF4 and c-MYC,so as to obtain the iPSCs from the FSHD patient.Then,the iPSCs were induced to differentiate into skeletal muscle cells.The characteristics of the iPSCs and skeletal muscle cells were evaluated by the optical genome mapping technolo-gy,karyotyping analysis,immunofluorescence staining,and real-time fluorescence quantitative PCR.Results The iPSCs from the FSHD patient were successfully obtained,which could express the markers of iPSCs.The karyotype and D4Z4 repeat unit of the iPSCs were consistent with that of the patient.The iPSCs could be induced to differentiate into skeletal muscle cells in vitro,which expressed the pathogenic gene DUX4 and its regulatory genes.Conclusion The PBMCs from one FSHD patient can be reprogrammed into iPSCs,which can be differentiated into disease-related myogenic progenitor cells and myotubes.This provides a useful cell model for in vitro studies of the pathogenesis of FSHD and a tool for the effective treatment of FSHD.

Spinal cord stimulation (SCS)-induced analgesia was characterized, and its underlying mechanisms were examined in a spared nerve injury model of neuropathic pain in rats. The analgesic effect of SCS with moderate mechanical hypersensitivity was increased with increasing stimulation intensity between the 20% and 80% motor thresholds. Various frequencies (2, 15, 50, 100, 10000 Hz, and 2/100 Hz dense-dispersed) of SCS were similarly effective. SCS-induced analgesia was maintained without tolerance within 24 h of continuous stimulation. SCS at 2 Hz significantly increased methionine enkephalin content in the cerebrospinal fluid. The analgesic effect of 2 Hz was abolished by μ or κ opioid receptor antagonist. The effect of 100 Hz was prevented by a κ antagonist, and that of 10 kHz was blocked by any of the μ, δ, or κ receptor antagonists, suggesting that the analgesic effect of SCS at different frequencies is mediated by different endorphins and opioid receptors.
Analgesics ; Animals ; Narcotic Antagonists/pharmacology* ; Neuralgia/therapy* ; Opioid Peptides ; Rats ; Receptors, Opioid/physiology* ; Receptors, Opioid, kappa ; Spinal Cord ; Spinal Cord Stimulation

The pain-relieving effect of acupuncture is known to involve primary afferent nerves (PANs) via their roles in signal transmission to the CNS. Using single-unit recording in rats, we characterized the generation and transmission of electrical signals in Aβ and Aδ fibers induced by acupuncture-like stimuli. Acupuncture-like signals were elicited in PANs using three techniques: manual acupuncture (MAc), emulated acupuncture (EAc), and electro-acupuncture (EA)-like peripheral electrical stimulation (PES). The discharges evoked by MAc and EAc were mostly in a burst pattern with average intra-burst and inter-burst firing rates of 90 Hz and 2 Hz, respectively. The frequency of discharges in PANs was correlated with the frequency of PES. The highest discharge frequency was 246 Hz in Aβ fibers and 180 Hz in Aδ fibers. Therefore, EA in a dense-disperse mode (at alternating frequency between 2 Hz and 15 Hz or between 2 Hz and 100 Hz) best mimics MAc. Frequencies of EA output >250 Hz appear to be obsolete for pain relief.

Objective: To understand the characteristics relating to the etiology and complications of hand, foot and mouth disease (HFMD) based on data from the pilot National Sentinel Surveillance (NSS) program so as to explore the feasibility, advantages and disadvantages of the NSS. Methods: Data were extracted from the NSS system, conducted in 11 provinces of China from November 2015 to October 2016. Characteristics regarding the etiology, complications of HFMD and factors related to the positive rates of HFMD specimens were analyzed under the logistic regression method by SPSS 20.0 software. Results: A total of 4 783 specimens were collected, including 3 390 from mild, 1 390 from severe and 3 from death cases. The overall positive rate was 81.43% (3 895/4 783). Other enteroviruses (non EV71/Cox A16 enteroviruses) appeared the major serotype (52.68%, 1 482/2 813) for mild infection of the disease while EV71 was for the severe cases (65.31%, 706/1 081). The serotype spectrum revealed by the pilot NSS was almost identical with the existing surveillance system. Other enteroviruses tended to infect younger children (χ²=130.17, P<0.001) than EV71 and Cox A16, in China. The multivariate logistic regression results showed that higher positive rate was associated with specimens which were collected from males, at children’ hospitals, in peak seasons, timely and in stools. The positive rates presented downwarding trends with the extension of the onset-sampling interval (χ²=14.47, P<0.001 in stool specimen; χ²=31.99, P<0.001 in throat swab; χ²=24.26, P<0.001 in anal swab). Aseptic meningitis, non-brainstem encephalitis and brainstem encephalitis appeared the top three complications of both EV71-associated and other enteroviruses-associated severe HFMD cases. Conclusions Factors as gender, season/place/timeliness of specimen collection, and types of hospital all appeared independently influenced the positive rates. NSS seemed feasible to be used as an alternative or supplement tool to the existing surveillance program in China.

OBJECTIVETo explore the anti-myeloma effect of suberoylanilide hydroxamic acid (SAHA) and on mouse myeloma cell line SP2/0 in vitro and in vivo and its mechanism.

METHODSThe inhibitory effect of SAHA on SP2/0 cells was measured by CCK-8 assay,and the apoptosis and cell cycle were analyzed by flow cytometry FACS. The protein expression of Caspase-3 and p53 of SP2/0 cells treated with SAHA were examined by Western blot. Annexin V/7-AAD double staining was performed to detect the apoptosis of SP2/0 induced by SAHA in vitro. SP2/0 cells (1×10) resuspended in 200 µl PBS were inoculated subcutaneously and intravenously into BALB/c mice, so as to establish aggressive or non-aggressive myeloma-bearing mouse models respectively. On day 3 after modeling, mice received SAHA or vehicle control treatment by intraperitoneal injection. The dose of SAHA was 60 mg/kg·d, 5 times a week for 3 weeks.

RESULTSIn SAHA-treated SP2/0 cells, the proliferation inhibition rate and apoptotic cells increased in a dose dependent manner. Also, SAHA significantly increased the ratio of cells in G phase and decreased in S phase. Molecular mechanisms of apoptosis and cell cycle arrest of SP2/0 induced by SAHA partly correlated with up-regulating the expression level of Caspase-3 and p53. In the non-aggressive myeloma-bearing mice, SP2/0 cells disappeared in peripheral blood after SAHA treatment. In the aggressive myeloma-bearing mice, inhibition of tumor growth and prolongation of the cell survival were observed after SAHA treatment.

CONCLUSIONSAHA inhibited SP2/0 cell proliferation, this effect associates with inducing apoptosis and cell cycle arrest, the mechanism of SAHA ralates partly with activating Caspase-3 and p53 pathway.

Animals ; Antineoplastic Agents ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Histone Deacetylase Inhibitors ; Hydroxamic Acids ; Mice ; Mice, Inbred BALB C ; Multiple Myeloma

Objective To investigate the clinical severity,etiological classification and risk factors of severe cases with hand,foot and mouth disease (HFMD).Methods A total of 1 489 records on severe and fatal HFMD cases reported to the national pilot surveillance system of HFMD were used to analyze the demographic,medical treatment,etiological classification of the cases.Treatment outcome related risk factors were also studied with multi-variable stepwise logistic regression method.Results Seven out of the 1 489 severe HFMD cases died of this disease.A total of 960 (72.9％) were under three years old and 62.9％ were male and most of the cases (937,62.9％) resided in rural areas.Among all the cases,494 (33.2％) went to seek the first medical assistance at the institutions of village or township level.Durations between disease onset and first medical attendance,being diagnosed as the disease or diagnosed as severe cases were 0(0-1) d,1 (0-2) d and 2 (1-4) d,respectively.In total,773 (51.9％) of the severe HFMD cases were diagnosed as with aseptic meningitis,260 (17.5％) with brainstem encephalitis,377 (25.3 ％) with non-brainstem encephalitis,6 (0.4％) with encephalomyelitis,1 (0.1％) with acute flaccid paralysis,4 (0.3％) with pulmonary hemorrhage/pulmonary edema and 68 (4.6％) with cardiopulmonary failure.Of the etiologically diagnosed 1 217 severe and fatal HFMD cases,642 (52.8％) were with EV71,other enterovirus 261 (21.5％),Cox A16 36 (3.0％),1 (0.1％) with both EV71 and Cox A16.However,277 (22.8％) showed negative on any pathogenic virus.Complication (Z=3.15,P=0.002) and duration between onset and diagnosed as severe cases (Z=3.95,P＜0.001) were shown as key factors related to treatment outcomes.Conclusions Most severe HFMD cases appeared in boys,especially living in the rural areas.Frequently seen complications would include aseptic meningitis,non-brainstem encephalitis and brainstem encephalitis.EV71 was the dominant etiology for severe and fatal cases.Early diagnosis and complication control were crucial,related to the treatment outcome of HFMD.

Precision medicine is one of the most important development in the future.In our study,the main contents of precision medication education were presented,and we aimed to provide a reference for clinical pharmacy educators.There are many factors affecting drug efficacy,and the factors could be divided into two categories of genetic and non-genetic factors.Genetic factors include genetic mutations,epigenetic modification,gene expression,miRNA etc.and non-genetic factors can be subdivided into drug-drug interaction,physiological factor,pathological factor,and other factors.Multiple knowledge was related to Precision medicine,such as pharmacogenetics,genetic testing,bioinformatics pharmacogenetics,therapeutic drug monitoring,pharmacometrics and so on.Take warfarin for example,we explained each factor effecting warfarin anticoagulant therapy.The commonly used databases for precise medication were also summarized.Related basic knowledge is the basis of precision medicine.In clinic,specific drug should be analyzed specifically.In order to constantly update knowledge,we should learn to use the common databases.

Objective To evaluate the effectiveness of sodium glycididazole combined with radiotherapy in the treatment of esophageal cancer.Methods RCTs were retrieved from EMbase,PubMed,Cochrane Library,CNKI,VIP,Wanfang database.RCTs about sodium glycididazole combined with radiotherapy in the treatment of esophageal cancer were included.The schedule for treatment group was sodium glycididazole combined with radiotherapy,and that for control group was radiotherapy only.The complete remission rate (CR),total remission rate and incidence of adverse reaction were the combined effect amount and they were evaluated by using Rev Man 5.3 statistical software.Results A total of 9 RCTs were included,which involved 854 patients.Meta-analysis showed that odds ratio of esophageal cancer for both treatments group and control group were 54.22％ (244 cases/450 cases) and 28.46％ (115 cases/404 cases) respectively.Toial remission rate for both groups were 96.44％ (434 cases/450 cases) and 80.41％ (324 cases/404 cases),with significant difference (all P ＜ 0.05).The incidences of adverse reactions were 49.42％ (86 cases/174 cases) in treatment group and 55.83％ (67 cases/120 cases) in control group which had no significant difference(P ＞0.05).Conclusion Short-term efficacy of sodium glycididazole combined with radiotherapy was better than that by radiotherapy alone for esophageal cancer.

Humans ; Liver Cirrhosis ; diagnosis ; mortality ; physiopathology ; Logistic Models ; Prognosis

OBJECTIVETo investigate the incidence of JAK2V617F gene point mutation in patients with myeloproliferatives diseases (MPD) and its clinical significance.

METHODSGenomic DNA from bone marrow and peripheral blood cells were extracted from 68 patients with MPD. Allele specific polymerase chain reaction was used to amplify the exon 12 of JAK2 gene which harbours V617F mutation. The PCR products were identified by DNA sequencing. JAK2V617F gene point mutation and its impact on peripheral blood cells were analyzed.

RESULTSThe incidence of JAK2V617F mutation in 68 patients with MPD was 65.28 %. The positive rate of JAK2V617F point mutation was 77.77 % in patients with PV (36/59), 56.52 % in patients with ET (23/59) and 44.44 % in patients with IMF (4/9). In all groups, the incidence of JAK2V617F point mutation in bone marrow and peripheral blood were equal. Patients with JAK2V617F mutation in PV group had higher counts of white blood cell and hemoglobin in peripheral blood than patients without JAK2V617F point mutation (P <0.05). Patients with JAK2V617F mutation in ET group had higher counts of white blood cell than those without JAK2V617F mutation (P <0.05); there was no significant difference in platelet count.

CONCLUSIONJAK2V617F point mutation can affect the hematologic features, which may be of diagnostic value for MDP with negative BCR-ABL gene.

Adolescent ; Adult ; Aged ; Amino Acid Substitution ; Base Sequence ; Female ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Myeloproliferative Disorders ; enzymology ; genetics ; Point Mutation ; Young Adult