Hand, Foot and Mouth Disease ; diagnosis ; therapy ; Health Education ; Humans

Adolescent ; Anemia, Aplastic ; blood ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Lymphocyte Subsets ; immunology ; Male ; Retrospective Studies ; Severity of Illness Index ; T-Lymphocytes, Cytotoxic ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

In order to construct generally efficient cell immortalization vector, pTP-hTERT, we modified the traditional piggyBac (PB) transposon using artificial synthesis, PCR and enzyme digestion. The modified vector contained the necessary transposon elements, a PB transposase expression cassette, a co-expression selectable element and a human telomerase reverse transcriptase (hTERT) expression cassette. The co-expression selectable element had two markers, enhanced green fluorescent protein (EGFP) gene and puromycin-resistance (Puro) gene, linked by porcine teschovirus-1 2A peptide (P2A). To validate the functionality of vector elements, we transfected pTP-hTERT into HEK293 cell, selected the positive cell clones and then conducted RT-PCR, Western blotting (WB) and Tail-PCR, methylene blue staining and statistic analysis on selected cells. The results of sequencing and cell culture show that the pTP-hTERT was constructed successfully and the positive cell could be selected by puromycin. The WB results, P2A cutting EGFP and Puro fusion protein with high efficiency, reflected the selectable element worked. The sequencing result of Tail-PCR confirmed the vector integrated into the genome through transposition. The results of methylene blue staining and statistic analysis indicated the clone of positive cells triggered by pTP-hTERT significantly increased (P < 0.01) compared with control group. The construction of pTP-hTERT provides an efficient tool for establishing immortalized cell lines and a demonstration for building other eukaryotic plasmids.
Cell Line ; DNA Transposable Elements ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Telomerase ; genetics ; Transfection

BACKGROUND:At present, spleen tyrosine kinase is the new target of studying and treating rheumatoid arthritis. OBJECTIVE:To study the influence of smal molecule tyrosine kinase inhibitor HL131078 on the inflammatory cel infiltration and cartilage destruction of the knee joint of mice with col agen-induced arthritis. METHODS:Forty DBA/1 mice were randomly and evenly divided into blank, model, positive and experimental groups. Col agen type II (CII) solution and Freund’s complete adjuvant (including mycobacterium tuberculosis) were injected into the mice of the latter three groups through the tail to establish mouse models of col agen-induced arthritis. At 2 weeks after the the first immunization with CII, the mice in the positive group were intragastrical y given R406 (10 mg/kg), once a day, for 28 consecutive days. The mice in the experimental group were intragastrical y given HL131078 (10 mg/kg), once per day, for 28 consecutive days. RESULTS AND CONCLUSION:Compared with the model group, the mean arthritis indexes of mice in the experimental and positive groups started to decline at 29 and 26 days. In the experimental group, the cartilage destruction of mouse knee joint was obviously reduced and the inflammatory cel infiltration in the knee joints was obviously reduced, which was close to that in the positive group. The results demonstrate that the smal molecule tyrosine kinase inhibitor HL131078 can effectively reduce inflammatory cel infiltration and cartilage destruction in the knee joints of mice with col agen-induced arthritis.

Objective To observe the effects of mechanical ventilation with different tidal volumes (Vt) on the pulmonary surfactant in dogs during cardiopulmonary resuscitation (CPR) and discuss the strategy of mechanical ventilation for CPR.Methods A total of 24 healthy,adult and mongrel canines were randomly (random number) divided into three groups,namely control group (A),low Vt group (B) and high Vt group (C).Canines without ventricular fibrillation in group A were not supported with mechanical ventilation,while canines in group B and group C were induced to get ventricular fibrillation (VF) with pacing catheter inserted through right external carotid vein into the right ventricle.CPR was initiated with mechanical ventilation with different Vt for 6 hours after ventricular fibrillation.The Vt set in group B was 6 ml/kg body weight and Vt set in group C was 20 ml/kg body weight.The arterial blood gas analyses were tested in different intervals.The levels of phosphatidylchline in bronchia alveolus lavage fluid (BALF) was assayed with high performance liquid chromatography (HPLC).The ratio of wet and dry weight and the morphological change scores of lung tissue were calculated.Results After ventricular fibrillation,pH,PaO2 in both groups B and C decreased and PaCO2 increased in comparison with those before VF (P ＜ 0.05),while pH,PaO2 and PaCO2 were improved after mechanical ventilation.These was statistical difference in PaCO2 between group B and group C 2 h after mechanical ventilation (P ＜ 0.05).There was difference in HCO3-between group B and group C 6 h after ROSC.The rate of successful restoration of spontaneous circulation (ROSC) and the 72 h survival rate were 75％ and 50％ respectively in group B,and 62.5％ and 40％ respectively in group C.There were no statistically different between two groups (P ＞ 0.05).The levels of phosphatidylchline in BALF of group B and C greatly decreased compared with the group A (All P ＜0.01),while phosphatidylcholine detected in the group C decreased much more than that in the group B (P ＜0.01).The W/D ratio and morphological change scores in the group C were higher than those in the group B (P ＜ 0.01).Conclusion Low tidal volume ventilation can reduce the loss of pulmonary surfactant and mitigate pulmonary injury.Therefore it may be a protective ventilation strategy after resuscitation.

Objective To explore the factors influencing pneumococcal vaccination uptake among the el-derly people in Guangzhou. Methods A survey by questionnaire was performed among 827 subjects aged 60 years or above and living in Guangzhou for five consecutive years. Chi-square test and multiple logistic regression analysis were applied to identify factors influencing pneumococcal vaccination uptake among the elderly. Results The positive factors for vaccination uptake among the elderly people included age of over 70 years (OR=1.677, 95%CI: 1.156 ~ 2.434), mental workers (OR = 1.837, 95%CI: 1.214 ~ 2.779), education background of over-three-year-course training (OR=1.769, 95%CI:1.039~3.012), and history of chronic diseases (OR=1.659, 95%CI:1.096~2.512) were positively associated with pneumococcal vaccination uptake. Monthly disposable income was not an influencing factor (OR=1.420, 95%CI: 0.895 ~ 2.251). Conclusion Strengthened publicity of pneumo-coccal vaccination among the elderly people and flexible measures tailored to the needs of different groups are rec-ommended in order to improve pneumococcal vaccination uptake among the elderly people.

Objective To analyze the efficacy of electric sacral neuromdulation and resiniferatoxin in patients with female overactive bladder.Methods 32 cases with IOAB female patients accepted percutaneous test sitmulation of the electric sacral nerves at S3 ,and treated by intravesical instillation with 100ml of 100nmol/L RTX.The effica- cy of voiding status were evaluated.The improvement of female patients life were evaluated comparing the rating of depression and anxiety.Results There were significant improvements in 32 cases in variables included the number of voiding,volume voided and signs every day and urgent uresis.In the rating of depression and anxiety,the patients improved a litter and still had stimulating symptom in urethra and bladder.Conclusion The treatment of IOAB with single administratoon of electric sacral neuromdulation and resiniferatoxin is effective,and can successfully im- prove the symptom with little side effects.

Objective To investigate the changes of sympathovagal balance and the effects of va- gus stimulation on sympathovagal balance in endotoxemia rats.Methods Twenty-four Spragne Dawley rats were randomly divided into four groups.The frequency domain of heart rate variability(HRV)com- ponent was analyzed at 0 min,2 ,4 and 6 hours after intravenous injection of lipopolysaccharide(LPS, 5mg/kg)or physiologic saline,and cervical vagal nerve was stimulated(5mv,2ms,1Hz,5 min lasted, 20 min interval)when LPS or physiologic saline was injected.The levels of Noepinephine(NE)and Ace- tylcholin(ACh)were measured in liver tissues.Results Normalized low frequency(LFnm),hormali- zed high frequency(HFnm),very low frequency(VLF),LF/HF values and liver ACh were significantly increased(P＜0.05)and the level of liver NE was significantly decreased (P＜0.05)after LPS admin- istration.Vagal nerve stimulation markedly increased HFnm but decreased LFnm,VLF,LF/HF values, and the liver ACh also significant increased(P＜0.05 ).Conclusion The results suggest that the ac- tivity of sympathetic and vagal nerve was increased during endotoxemia,but the sympathetic activity was more excitable than that of vagal nerve.Vagal nerve stimulation increased the tone of vagus nerve while the tone of sympathetic nerve was decreased in this study.This may be beneficial for anti-inflammatory activity of vagal nerve.

[Objective] To investigate the effect of Shangke Quyu Tablet (a formula modified from Tao Hong Siwu Decoction) in treating acute traumatic swelling and pain of soft tissue. [Methods] Two hundred and forty-six cases of traumatic injury in four limbs, which were classified to Qi-stagnation and blood-stasis syndrome, were randomized to two groups. After given fracture reduction and dislocation fixation, group A (n = 164) was treated with self-prepared Shangke Quyu Tablet and group B with Tongxuekang Capsules (n = 82) . Therapeutic effect was evaluated after 10 days of treatment. [Results] In group A, 116 cases were markedly improved, 30 much improved, 14 improved and 4 ineffective; in group B, 44 cases were markedly improved, 18 much improved, 16 improved and 4 ineffective. The therapeutic effect in group A was better than that in group B (P

Objective To observe clinical efficacy of Qilin Pills combined with Three-Spot Caressing in the treatment of acquired non-consolidated kidney qi premature ejaculation (PE). Methods Totally 129 acquired non-consolidated kidney qi PE patients were randomly divided into TCM group, Fangzhognshu group and combination group. TCM group was given Qilin Pills, 6 g each time, three times a day, orally. Fangzhognshu group was guided to conduct Three-spot Caressing during sexual intercourse, no less than twice a week. Combination group was given Qilin Pills and Three-spot Caressing with the same method as Fangzhognshu group. The treatment course for each group lasted for 8 weeks. Before and after treatment of 4 weeks, 8 weeks and 4 weeks after with drawal, the intravaginal ejaculatory latency time (IELT), Chinese Index of Sexual Function for Premature Ejaculation-5 (PECI-5) scores and TCM syndrome scores (TCMS) were compared. Results Compared with before treatment, IELT increased in three groups at different time points after treatment (P<0.05). In the combination group, IELT was superior to the other two groups (except for TCM group after 4-week treatment), with statistical significance (P<0.05). Compared with before treatment, PECI-5 scores improved in three groups (except for Q6 and Q7 in Fangzhongshu group after 4-week treatment) at different time point after treatment (P<0.05). The improvement of Q5, Q6 and Q7 scores in PECI-5 of combination group was significantly better than that in the other two groups at 8-week treatment and 4-week after treatment (P<0.05). Compared with before treatment, TCMS in TCM group and combination group were improved significantly (P<0.05), PE and loss of libido score improved significantly in Fangzhongshu group (P<0.05);mental malaise and move spontaneous perspiration were significantly improved at 8-week treatment and 4 weeks after treatment (P<0.05). The improvement in TCMS was significantly better in combination group than that in the other two groups at 8-week treatment and 4 weeks after treatment (P<0.05). Conclusion Qilin Pills combined with Three-Spot Caressing has definite and long-term efficay for the treatment of acquired non-consolidated kidney qi PE.