ObjectiveTo construct a recombinant virus expressing tick-borne encephalitis virus(TBEV) prM-E protein using vesicular stomatitis virus(VSV) as a vector, and to identify it, so as to provide a basis for the research of TBEV vaccines based on VSV vector.MethodsThe prM-E gene of the TBEV Senzhang strain was inserted between the G and L genes of the VSV vector. The recombinant virus was rescued by co-transfecting BHK-21 cells infected by poxvirus containing T7 RNA polymerase with helper plasmids expressing VSV N, P, L, and G proteins. The supernatant was collected, and the recombinant virus rVSV-TBEVprM-E stably expressing prM-E was obtained after multiple passages. Western blot, immunofluorescence assay(IFA) and RT-PCR were used to identify the expression of prM-E protein and gene of rVSV-TBEVprM-E in cells. The viral titers of rVSV-TBEVprM-E at different time points were determined by plaque assay and the growth curve was plotted.ResultsTBEV prM-E gene was successfully inserted into the genome of recombinant virus rVSV-TBEVprM-E, and the expression of prM-E protein in BHK-21 cells was detectable. After serial passages, rVSV-TBEVprM-E achieved a viral titer of 6. 75 × 10~5 PFU/mL.ConclusionA recombinant virus rVSV-TBEVprM-E expressing prM-E protein was successfully constructed, which lays a solid experimental foundation for the related research of TBEV.

Objective To explore the distribution of common TCM syndromes and symptoms of insomnia;To prepare for the construction of the theoretical framework and item pool of syndrome diagnosis and efficacy evaluation scale.Methods TCM guideline standards of insomnia,textbooks and journals over the years were retrieved,the information of TCM syndromes,syndrome elements and symptoms was extracted,the guideline textbook and journal database were established,and descriptive statistics,association rules,systematic clustering,factor analysis,potential categories and implicit structure analysis were carried out.Results Totally 116 guide standards and textbooks over the years were included,and 454 articles of journals were included.The high-frequency symptoms accounted for≥3%of the guide textbooks and journal databases were 87 and 79 categories,respectively,and the cumulative proportion was 87.48%and 87.75%,respectively.According to the analysis results,five common TCM syndromes and their characteristic symptom classification of insomnia were finally deduced.According to the frequency/person time distribution,they were heart and spleen deficiency syndrome,yin deficiency and fire hyperactivity syndrome,liver fire disturbing heart syndrome,phlegm heat disturbing heart syndrome,heart and gallbladder qi deficiency syndrome.Conclusion There are five common TCM syndromes of insomnia,and the characteristic symptoms of each TCM syndrome provide a reference source for the theoretical framework of syndrome diagnosis and efficacy evaluation scale and the establishment of item pool.

Objective To explore the distribution of common TCM syndromes and symptoms of insomnia;To prepare for the construction of the theoretical framework and item pool of syndrome diagnosis and efficacy evaluation scale.Methods TCM guideline standards of insomnia,textbooks and journals over the years were retrieved,the information of TCM syndromes,syndrome elements and symptoms was extracted,the guideline textbook and journal database were established,and descriptive statistics,association rules,systematic clustering,factor analysis,potential categories and implicit structure analysis were carried out.Results Totally 116 guide standards and textbooks over the years were included,and 454 articles of journals were included.The high-frequency symptoms accounted for≥3%of the guide textbooks and journal databases were 87 and 79 categories,respectively,and the cumulative proportion was 87.48%and 87.75%,respectively.According to the analysis results,five common TCM syndromes and their characteristic symptom classification of insomnia were finally deduced.According to the frequency/person time distribution,they were heart and spleen deficiency syndrome,yin deficiency and fire hyperactivity syndrome,liver fire disturbing heart syndrome,phlegm heat disturbing heart syndrome,heart and gallbladder qi deficiency syndrome.Conclusion There are five common TCM syndromes of insomnia,and the characteristic symptoms of each TCM syndrome provide a reference source for the theoretical framework of syndrome diagnosis and efficacy evaluation scale and the establishment of item pool.

Objective:To investigate the relationship between the number of epithelial progenitor cells(EPCs)in peripheral blood and blood parameter values in order to clarify the relationship between peripheral blood EPCs and the length of survival in elderly individuals aged 75 and beyond.Methods:Analysis was conducted on 114 individuals ≥75 years recruited from the Department of Geriatrics, Tianjin Medical University General Hospital in 2008.Clinical data were collected with December 31, 2020 as the end of the follow-up.Associations between peripheral blood EPC numbers, blood parameter values and all-cause mortality were analyzed.Results:In low and high EPC groups based on orthogonal partial least squares-discriminant analysis(OPLS-DA), D-Dimer was found to be a potential classification marker[the variable importance in projection(VIP)=4.750], out of a panel of blood tests.Blood parameters such as red blood cells(RBC), hemoglobin(HGB), hematocrit(HCT), aspartate aminotransferase(AST), gamma-glutamyl transferase(GGT), lactate dehydrogenase(LDH)and D-Dimer showed statistically significant differences between groups with different levels of EPCs(all P<0.05). Blood EPC numbers were positively correlated with RBC, HGB and HCT and negatively correlated with GGT and D-Dimer in older individuals( r=0.364, 0.343, 0.336, -0.312, -0.312, P<0.05 for all). The number of circulating EPCs had a significant positive correlation with survival time in older individuals( r=0.234, P=0.017). Based on the data of the long-term(12-years)follow-up, Logistic regression analysis suggested that the number of EPCs, age, and HCT were associated with all-cause mortality events in older individuals( OR=0.971, 1.585, 1.231, P=0.013, 0.012, 0.029). Conclusions:EPCs in the peripheral circulation may be involved in the progression of various conditions such as anemia, cardiac, hepatic and renal injury, and coagulation.The number of circulating EPCs may influence the survival of elderly individuals ≥75 years.Age and HCT may be risk factors for all-cause mortality events, whereas the number of EPCs may be a protective factor.

Ommaya reservoir implantation is generally used in the treatment of hydrocephalus and intraventricular drug administration. Ommaya reservoir implantation in the subarachnoid space of the spinal cord for the intrathecal drug administration has not been carried out in China, and only several reports can be retrieved from PubMed. About 60%-90% of untreated patients with spinal muscular atrophy type 2 (SMA2) who survive to adulthood often have complex scoliosis and joint deformities. Nusinersen is an effective drug for the treatment of SMA2. And the route of administration is intrathecal injection, which is difficult for patients with severe scoliosis. This article summarizes the process of Ommaya reservoir implantation and postoperative drug administration in a patient with complex scoliosis type SMA2, which provides a new method for clinical treatment of this disease.

Objective: To investigate the incidence and trend of short-term outcomes among preterm infants born <34 weeks' gestation. Methods: A secondary analysis of data from the standardized database established by a multicenter cluster-randomized controlled study "reduction of infection in neonatal intensive care units (NICU) using the evidence-based practice for improving quality (REIN-EPIQ) study". This study was conducted in 25 tertiary NICU. A total of 27 192 infants with gestational age <34 weeks at birth and admitted to NICU within the first 7 days of life from May 2015 to April 2018 were enrolled. Infants with severe congenital malformation were excluded. Descriptive analyses were used to describe the mortality and major morbidities of preterm infants by gestational age groups and different admission year groups. Cochran-Armitage test and Jonckheere-Terpstra test were used to analyze the trend of incidences of mortality and morbidities in 3 study-years. Multiple Logistic regression model was constructed to analyze the differences of outcomes in 3 study-years adjusting for confounders. Results: A total of 27 192 preterm infants were enrolled with gestational age of (31.3±2.0) weeks at birth and weight of (1 617±415) g at birth. Overall, 9.5% (2 594/27 192) of infants were discharged against medical advice, and the overall mortality rate was 10.7% (2 907/27 192). Mortality for infants who received complete care was 4.7% (1 147/24 598), and mortality or any major morbidity was 26.2% (6 452/24 598). The incidences of moderate to severe bronchopulmonary dysplasia, sepsis, severe intraventricular hemorrhage or periventricular leukomalacia, proven necrotizing enterocolitis, and severe retinopathy of prematurity were 16.0% (4 342/27 192), 11.9% (3 225/27 192), 6.8% (1 641/24 206), 3.6% (939/25 762) and 1.5% (214/13 868), respectively. There was a decreasing of the overall mortality (P<0.001) during the 3 years. Also, the incidences for sepsis and severe retinopathy of prematurity both decreased (both P<0.001). However, there were no significant differences in the major morbidity in preterm infants who received complete care during the 3-year study period (P=0.230). After adjusting for confounders, infants admitted during the third study year showed significantly lower risk of overall mortality (adjust OR=0.62, 95%CI 0.55-0.69, P<0.001), mortality or major morbidity, moderate to severe bronchopulmonary dysplasia, sepsis and severe retinopathy of prematurity, compared to those admitted in the first study year (all P<0.05). Conclusions: From 2015 to 2018, the mortality and major morbidities among preterm infants in Chinese NICU decreased, but there is still space for further efforts. Further targeted quality improvement is needed to improve the overall outcome of preterm infants.
Bronchopulmonary Dysplasia/epidemiology* ; Gestational Age ; Humans ; Infant ; Infant Mortality/trends* ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases/epidemiology* ; Patient Discharge ; Retinopathy of Prematurity/epidemiology* ; Sepsis/epidemiology*

Anti-Bacterial Agents/therapeutic use* ; Carbapenem-Resistant Enterobacteriaceae ; Carbapenems ; Child ; Cross Infection/epidemiology* ; Enterobacteriaceae Infections/epidemiology* ; Humans ; Microbial Sensitivity Tests ; Risk Factors ; beta-Lactamases

Objective:To study the inhibitory effect of deoxyschizandrin on the growth of brain glioma C6 cells, and to explore its mechanism.Methods:The rat glioma C6 cells were cultured and divided into control group,50, 100,and 200 mg·L-1 deoxyschizandrin groups.The proliferation rates of C6 cells were examined by MTT assay;the changes of cell cycles were examined by flow cytometry;the expression levels of CyclinD1,Bax,Bcl-2 and Caspase-3 proteins in supernant were detected by ELISA assay. Results:Compared with control group, the proliferation rates at 24 and 48 h in 50,100,and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P <0.01),and the proliferation rates at 72 h in 100 and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P < 0.05 or P < 0.01 ). Compared with control group, the percentage of cells at SubG1 phase in 200 mg·L-1 deoxyschizandrin group was increased (P < 0.05 ), and the percentage of cells at S phase was decreased (P <0.05).Compared with control group,the expression levels of CyclinD1 in 100 and 200 mg· L-1 deoxyschizandrin groups were decreased (P < 0.01 );the expression levels of Bax protein in deoxyschizandrin groups were increased (P < 0.05 or P < 0.01 ), and the expression level of Bcl-2 protein in 200 mg · L-1 deoxyschizandrin group was decreased (P < 0.01 ), and the Bax/Bcl-2 value in deoxyschizandrin groups were increased (P < 0.01 ); the expression level of Caspase-3 protein in 200 mg · L-1 deoxyschizandrin group was increased (P < 0.01 ).Conclusion:Deoxyschizandrin could inhibit the growth of glioma cells through down-regulating the expression levels of CyclinD1 protein and up-regulating the expression levels apoptotic factors Bax and Bcl-2.

10.3969/j.issn.2095-4344.2013.23.010

OBJECTIVETo investigate the incidence of local reactions (LRs) and systemic reactions (SRs) of subcutaneous immunotherapy (SCIT) and to analyze the potential risk factors of such reactions in Chinese population.

METHODThis is a retrospective study on 234 dust mite sensitized patients with allergic rhinitis and asthma who received allergen immunotherapy in our hospital from 2003 to 2010. Chart review was conducted to capture clinical data of reactions to immunotherapy. Parameters included signs and symptoms, the onset of reaction, and interventions in treating such reactions, particularly, the administration of epinephrine (EPI) and adjustment of vaccine dosage due to LRs and SRs.

RESULTThe 234 patients received a total of 7679 injections. Among them, 4973 LRs (64.8%) and 235 SRs (3.1%) were observed in 67 patients (28.6% of all patients). SRs included respiratory symptoms (205 events, 88.4%) and cutaneous symptoms (31.5%). Of the total of 235 SR events, 212 (90.2%) were presented as mild SRs and 23 (9.8%) were in severe SR category (grade III and grade IV, EAACI grading system). Overall, severe SRs accounted for 0.3% of total injections. Seventeen of the 23 SR events required epinephrine treatment (0.2% of total injections). Of the 67 patients, 61 completed the course of treatment after dose adjustment; 36 patients had their doses decreased prior to further advancing to target dose. Nineteen subjects tolerated splitting two injections at 30 minutes interval. Six patients advanced the dose based on protocol and another 6 had to stop immunotherapy. Most of the SRs (77.4%) occurred during the maintenance phase of immunotherapy. The levels of TIgE, SIgE D1 and SIgE D2 were found to be significantly higher in patients with SRs comparing to patients without SRs (P < 0.05). SRs more commonly occurred in patients with age less than 14 years than their older counterparts (95.5% vs. 85.6%, OR = 3.58, 95%CI = 1.040 - 12.322, P < 0.01). The incidence of SRs were significantly higher in asthma patients who received SCIT than non-asthma patients (OR = 2, 95%CI = 1.136 - 4.624).

CONCLUSIONOur study suggests that risk factors of SRs include maintenance phase (higher allergen vaccine doses), patients with asthma, age of less than 14 years, higher levels of TIgE, and SIgE D1 and SIgE D2. Effective management includes proper dose adjustment, splitting doses into 2 injections at 30 min apart, and strictly following immunotherapy indications.

Adolescent ; Adult ; Animals ; Antigens, Dermatophagoides ; administration & dosage ; immunology ; Asthma ; immunology ; therapy ; Child ; Child, Preschool ; Desensitization, Immunologic ; adverse effects ; methods ; Female ; Humans ; Hypersensitivity, Immediate ; epidemiology ; etiology ; therapy ; Injections, Subcutaneous ; Male ; Middle Aged ; Mites ; immunology ; Retrospective Studies ; Rhinitis, Allergic, Perennial ; immunology ; therapy ; Risk Assessment ; Treatment Outcome ; Young Adult