We constructed a recombinant Bacillus Calmette‐Guerin(BCG)‐TSOL18 vaccine of Taenia solium and observed the expression of the TSOL18 gene in BCG .The TSOL18 gene of Taenia solium was obtained from the recombinant plasmid pGEX‐TSOL18 by digestion method and cloned into Escherichia coli (E .coli)‐mycobacterium shuttle plasmid pMV261 to con‐struct the recombinant plasmid pMV261‐TSOL18 of Taenia solium ,and the recombinant plasmid was identified by restriction enzyme digestion ,PCR and DNA sequencing .Then ,the recombinant plasmid was transformed into BCG by electroporation to construct the recombinant BCG‐TSOL18 vaccine of Taenia solium ,and the vaccine was identified by PCR .The expression of the TSOL18 gene in BCG was identified by SDS‐PAGE and Western blot .The 393 bp TSOL18 gene fragment was successfully obtained by restriction enzyme digestion .Restriction enzyme digestion ,PCR and DNA sequencing suggested that the recombi‐nant plasmid pMV261‐TSOL18 of Taenia solium was successfully constructed .PCR confirmed that the recombinant plasmid pMV261‐TSOL18 of Taenia solium was successfully transformed into BCG ,suggesting that the recombinant BCG‐TSOL18 vaccine of Taenia solium was successfully constructed .SDS‐PAGE showed that the relative molecular mass (Mr) of TSOL18 target protein was approximately 14 .7 kD .Results of western blot showed the TSOL18 target protein could be recognized by rabbit antiserum or cysticercosis swine serum .The recombinant BCG‐TSOL18 vaccine of Taenia solium was successfully con‐structed .The TSOL18 gene of Taeniasolium was successfully expressed in BCG and the expressed TSOL18 recombinant pro‐tein had specific antigenicity .This result would lay a foundation for further study of the vaccine .

Objective To explore the effect of suledexide on renal injury and podocalyxin expression of podocytes in STZ diabetic desoxycorticosterone acetate (DOCA)-hypertensive rats. Methods Wistar rats were subjected to subcutaneous injection of streptozotocin(STZ), followed by uninephrectomy and subcutaneous administration of DOCA. Diabetic and hypertensive rats were randomly allocated to treatment with sulodexide or a combination of sulodexide and telmisartan for 8 weeks. Blood pressure (BP), 24-hour urinary albumin were measured every 2 weeks. Blood and urinary samples were collected to detect biochemical indexes of plasma and urinary β-acetyl-β-D-glucosaminidase (NAG) at the end of the study. Immunohistochemistry (IHC), RT-PCR and Western blot were performed to examine the expression and distribution of podocalyxin. Results STZ +DOCA-treated rats progressively developed hypertension, albuminuria and hyperglycemia. Hyperlipidemia and hypoinsulinemia were found in diabetic and hypertensive rats compared with controls. Albuminuia was significantly reduced in sulodexide group at week 8 and sulodexide plus telmisartan group at week 6 and week 8. Blood pressure decreased in sulodexide plus telmisartan group. No significant effects on lipid and glucose metabolism were observed in all treated groups. Histopathological index increased in STZ+DOCA-treated rats, but was significantly lower in sulodexide group as well as sulodexide plus telmisartan group. The number of podocytes on glomerular cross-section of the four groups were comparable. Segmental loss and down-regulation of podocalyxin were detected in STZ+DOCA-treated rats, which were greatly attenuated by sulodexinde, meanwhile, combination treatment preserved more podocalyxin expression in glomeruli than sulodexide monotherapy. Conclusion Sulodexide effectively reduces albuminuria, prevents loss of podocyte podocalyxin and alleviates renal damage in STZ diabetic DOCA-hypertensive rats.

To investigate the weight reduction of hydatid cyst and level of splenocyte subsets in mice immunized with recombinant Bifidobacteria bifidum (Bb) Bb-Eg95-EgA31 vaccine of Echinococcus granulosus (Eg) and challenged with Eg protoscoleces,BALB/c mice were immunized with the vaccine subcutaneously,intramuscularly,intranasally or orally.Then the mice were challenged with 50 Eg protoscoleces on 8~(th) week after vaccination and killed on 25~(th) week after infection.The weight of hydatid cyst was measured and the reduction of the weight was calculated.The spleens were collected for splenocytes preparation.CD_4~+ and CD_8~+ T cells were determined by flow cytometry(FCM),and the blank vector,Bb or MRS were set as control.The hydatid cyst weight was reduced by 45.33%,41.33%,70.67% and 62.67% respectively for the 4 immunization groups.Number of CD_4~+ and CD_8~+ subsets in the immunization groups was more than that in the control group.CD_4~+ and the ratio of CD_4~+/CD_8~+ in the intranasal or oral immunization group was much higher than that in the subcutaneous or intramuscular immunization group.It's concluded that CD_4~+ and CD_8~+T cells may play an important role in the protection induced by recombinant Eg95-EgA31 vaccine of Eg.In addition,intranasal and oral vaccinations could be the better ways for immunization.

Objective: Through the bibliometrics analysis, to understand the research status quo and trend of the application of gamification in health management. Methods: Based on the Web of Science Core Collection, We retrieved the article, review and proceedings paper published from 1900 to 2018 and analyzed literature retrieved in term of document types, publication years, authors, and funding agencies. In addition, VOSviewer1.6.9 was used for visualization analysis of keywords and countries/regions. Results: A total of 364 pieces of literature were retrieved, of which 177 (48.63%) pieces were article. Since 2013, the number of literature has shown an upward trend at a rapid speed. However, only 16.48% of the literature were published by high productivity authors, which means a core group of authors has not yet formed in this research direction. The United States published the most literature, 89 pieces. Seven of the top ten funding agencies by volume were belong to the United Kingdom. However, only 10 pieces of literature were published by China. A total of 55 high-frequency keywords were included in this study, forming 5 research topics: physical activities management of adolescent based on mobile devices; internet-based behavior and mental health management for older-adults; design and application of fitness games; intervention and promotion of youth health behavior; the application of mobile health in adult self-management. The new high frequency keywords included older-adults, adults, mental health etc. Conclusion The application of gamification in health management is in its infancy, but it develops rapidly. European and American countries are in a leading position, while China is relatively backward. The research topics tend to be diversified and gamification is initially applied to the older-adults, adults and mental health management. In the future, the application scope of gamification in health management should be further expanded and effectively combined gamification with mHealth.

Objective To dynamically observe humoral and cellular immune responses induced in mice by immunization with a recombinant BCG-TSOL18 vaccine of Taenia solium.Methods Totally 80 Kunming mice were divided into 4 groups by using random number table according to body mass, 20 mice in each group: rBCG-TSOL18 intraperitoneal injection group [mice were vaccinated with 5 × 106 colony forming units (CFU) recombinant BCG-TSOL18 vaccine of Taenia solium through intraperitoneal injection], rBCG-TSOL18 intragastric administration group(mice were vaccinated with 4 × 108 CFU recombinant BCG-TSOL18 vaccine of Taenia solium through intragastric administration), BCG control (mice were vaccinated with 5 × 106 CFU BCG through intraperitoneal injection), PBS control (mice were vaccinated with PBS through intraperitoneal injection).Zero, 2, 4, 6 and 8 weeks after immunization, eye blood was collected and serum w as separated.Levels of specific IgG and IgG2a were detected by enzyme linked immunosorbent assay (ELISA).Proliferation level of spleen lymphocytes was detected by CCK-8.Levels of interleukin-2 (IL-2) and IL-4 were determined by ELISA.Results The level of specific IgG in rBCG-TSOL18 intraperitoneal injection group and rBCG-TSOL18 intragastric administration group increased from 2 to 8 weeks, and reached the highest level by the 6th week (0.310 ± 0.022, 0.356 ± 0.026).Compared with 0 week in the same group, BCG and PBS control group of the same time periods (0.054 ± 0.005, 0.057 ± 0.006, 0.093 ± 0.014, 0.085 ± 0.010), there were statistically significant differences (all P ＜ 0.05).The level of specific IgG2a increased from 2 to 8 weeks, and reached the highest level by the 6th week (0.965 ± 0.031, 1.144 ± 0.049).Compared with 0 week in the same group, BCG and PBS control group of the same time periods (0.102 ± 0.014, 0.093 ± 0.012, 0.115 ± 0.012, 0.103 ± 0.013), there were statistically significant differences (all P ＜ 0.05).The proliferation level of spleen lymphocytes increased from 2 to 8 weeks, and reached the highest level by the 6th week (0.524 ± 0.032, 0.755 ± 0.016).Compared with 0 week in the same group, BCG and PBS control group of the same time periods (0.301 ± 0.018, 0.305 ± 0.020, 0.362 ± 0.033, 0.334 ± 0.027), there were statistically significant differences (all P ＜ 0.05).The levels of IL-2 and IL--4 in spleen lymphocyte culture supernatant increased from 2 to 8 weeks and from 4 to 6 weeks, respectively, reached the highest level by the 6th and 4th weeks, respectively [(1 665.41 ± 33.93), (1 785.11 ± 42.39)ng/L and (281.62 ± 23.79), (357.95 ± 42.57)ng/L].Compared with 0 week in the same group, BCG and PBS control group of the same time periods [(1 411.63 ± 20.54), (1 405.12 ± 21.42),(1 455.20 ± 25.03), (1 434.47 ± 17.47)ng/L and (190.17 ± 11.01), (196.96 ± 14.00), (200.51 ± 30.79), (189.64 ±40.90)ng/L], there were statistically significant differences (all P ＜ 0.05).These indexes mentioned above were statistically significantly different (all P ＜ 0.05) between rBCG-TSOL18 intragastric administration group and rBCG-TSOL18 intraperitoneal injection group.Conclusion The recombinant BCG-TSOL18 vaccine of Taenia solium might induce mice to produce humoral and cellular immune responses, and the effect of intragastric administration is better than that of intraperitoneal injection.

AIM:To observe the Toll-like receptor 9 (TLR9) activation in microglia BV-2 cells after oxygen-glucose deprivation and reoxygenation ( OGDR) , and its effects on neuronal apoptosis.METHODS:The BV-2 cell super-natants were collected after the corresponding treatment and added to mouse primary cortical neurons after OGDR for 4 h, followed by normal culture for 24 h.The cells were divided into normal BV-2 group, NC-siRNA group, TLR9-siRNA group, OGDR group, OGDR+NC-siRNA group, OGDR+TLR9-siRNA group and control group (without adding BV-2 cell supernatant) .The changes of the neuronal morphology were observed under an inverted phase-contrast microscope, and the neuronal apoptosis was detected by TUNEL.The protein expression of cleaved caspase-3 was detected by Western blot-ting.RESULTS:After OGDR, the axon turned thin, twisted and broken, and neuronal swelling, decrease in refraction and vacuolar degeneration were observed.The green-stained apoptotic bodies in the neurons in all groups were positive. Compared with control group, the caspase-3 protein levels in other groups were increased.Compared with the normal BV-2 group, the caspase-3 protein in OGDR group and TLR9-siRNA group was increased.Compared with OGDR+TLR9-siRNA group, the caspase-3 protein in TLR9-siRNA group and OGDR group was decreased.CONCLUSION: After OGDR, TLR9 activation in BV-2 cells induces neuronal apoptosis with the increase in caspase-3 protein level.Inhibition of TLR9 expression reduces neuronal damage.

Objective To study the protective immune responses induced by recombinant Bifidobacterium (Bb)-TSO45W-4B-TSOL18 vaccine of Taenia solium (T.solium) in domestic pigs challenged with T.solium eggs.Methods Twenty healthy 40 days old domestic pigs were divided into five groups by random number table according to body weight (15 kg):rBb-TSO45W-4B-TSOL18 vaccine group,rBb-TSO45W-4B vaccine group,rBb-TSOL18 vaccine group,blank vector control group and MRS control group.The content of vaccine in each vaccine group was 1 × 1011 CFU.A total of two immunization times was conducted,once every two weeks.Pigs were challenged with T.solium eggs 4 weeks after the last immunization and killed 3 months after infection.The cysticercus was counted and the reduction of the cysticercus was calculated.Blood was collected to separate sera and prepare peripheral blood lymphocytes (PBMC).The levels of IgG,IgG1 and IgG2a in sera were determined by enzyme linked immunosorbent assay (ELISA).The level of PBMC proliferation was tested using methyltetrazolium (MTT) assay.The levels of interleukin (IL)-2,interferon (IFN)-γ,IL-4 and IL-10 in PBMC culture supernatant were detected using ELISA.Results The reduction of cysticercus was 83.09％,71.36％ and 74.85％ in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups,respectively.The differences of IgG,IgG1,IgG2a levels in sera between groups were statistically significant (F =132.348,106.336,596.091,all P ＜0.05).The levels of IgG and IgG2a in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups [(366.81 ± 3.84),(334.94 ± 11.65),(333.52 ± 11.09),(87.74 ± 0.95),(84.48 ± 0.80),(84.30 ± 1.09)mg/L] were higher than those of the MRS control group [(245.94 ± 8.81),(62.61 ± 0.84)mg/L,all P ＜0.05].The levels of IgG1 in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups [(26.55 ± 1.06),(33.24 ± 1.92),(32.60 ± 1.94)mg/L] were lower than those of the MRS control group [(42.78 ± 0.87)mg/L,all P ＜0.05].The differences of IL-2,IFN-γ,IL-4 and IL-10 levels in PBMC original culture supernatant between groups were statistically significant (F =139.522,1 053.102,769.097,962.298,all P ＜0.05).The levels of IL-2 and IFN-γ in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups [(212.24 ± 3.12),(205.91 ± 3.18),(205.85 ± 4.35),(28.42 ± 0.28),(25.56 ± 0.28),(25.71 ± 0.35)ng/L] were higher than those of the MRS control group [(174.19 ± 2.14),(17.69 ± 0.28)ng/L,all P ＜0.05],while the levels of IL-4 and IL-10 [(40.45 ± 0.36),(41.38 ± 0.70),(41.52 ± 0.19),(71.45 ± 0.83),(73.38 ± 0.70),(74.77 ± 0.41)rig/L] were lower than those of the MRS control group [(52.57 ± 0.29),(94.82 ± 0.45)ng/L,all P ＜0.05].The differences of PBMC proliferation levels between groups were statistically significant (F =56.318,P ＜0.05).The PBMC proliferation levels in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups (0.543 ± 0.074,0.481 ± 0.028,0.530 ± 0.053) were higher than those of the MRS control group (0.242 ± 0.053,all P ＜0.05).Conclusions Recombinant Bb-TSO45W-4B-TSOL18 vaccine of T.solium could induce certain protection in domestic pigs.Type Th1 immune response may play an important role in induction of protective immunity.

Objective To study the protective effect of a mixed recombinant Bifidobacterium (Bb) vaccine of Taenia solium in piglets. Methods Healthy piglets of 40 days old were randomly divided into experimental group and control group, 4 in each group. Experimental group was given 1011 CFU rBb-TSO45W-4B vaccine and rBb-TSOL18 vaccine (in the ratio of 1 : 1). Control group was given Bb liquid medium (MRS). A total of two times of immunization were conducted, once for every two weeks. At different time points after immunization, the serum was separated from precaval vein blood to detect the level of IgG by enzyme linked immunosorbent assay (ELISA). Piglets were challenged with Taenia solium eggs on the 4th week after the last immunization and killed 3 months after infection. The cysticerci were separated to count and calculate the reduction rate of cysticerci. Blood from precaval vein was collected to separate serum and prepare peripheral blood lymphocytes (PBMC). The levels of IgG,IgG1 and IgG2a in serum sampl es and the levels of interleukin ( IL )-2 , interferon (IFN)-γ, IL-4 and IL-10 in PBMC culture supernatant were determined by ELISA. The level of PBMC proliferation was tested using methyltetrazolium (MTT) assay. Results In experimental group, the level of serum IgG increased from the 2nd to the 8th weeks after immunization, and reached the highest level on the 4th week after immunization (mg/L:270 . 64 ± 1 . 94 vs 207.74 ± 2.24, t=42.479, P<0.05). Reduction rate of cysticercus was 80.48%. Compared with control group, the levels of IgG and IgG2a in serum were significantly increased, while the level of IgG1 was significantly decreased (mg/L: 364.15 ± 11.52 vs 245.94 ± 8.81, 89.74 ± 1.13 vs 62.61 ± 0.84, 20.52 ± 1.00 vs 34.11 ± 0.65, t=16.303, 38.579, - 22.772, P < 0.05). The levels of IL-2 and IFN-γ in PBMC culture supernatant were significantly increased, while the levels of IL-4 and IL-10 were significantly decreased (ng/L:215.24 ± 3.31 vs 174.19 ± 2.14, 28.21 ± 0.27 vs 17.69 ± 0.28, 40.35 ± 0.34 vs 52.57 ± 0.29, 71.34 ± 0.36 vs 94.82 ± 0.45, t =20.839, 53.623,-54.743,- 81.266, P<0.05). The level of PBMC proliferation was significantly increased (0.620 ± 0.051 vs 0.242 ± 0.053, t=10.259, P<0.05). Conclusions It is concluded that the mixed rBb vaccine of Taenia solium might give piglets a certain protection. Th1 type immune response plays an important role in the protection.

Objective To explore the clinical and electrophysiological features and pulmonary function of 8 Chinese patients with Kennedy's disease (KD) and to enhance the understanding for the disease.Methods Eight patients with KD,admitted to out hospital from July 2010 to December 2013 and confirmed by gene examination,were chosen in our study; their clinical and electrophysiological features,and laboratory characteristics and pulmonary function were compared with those in the healthy volunteers.Results The average age of onset in the 8 patients was (45.13±17.47) years,and the average age of diagnosis was (55.63±12.11) years.The most common complaint was leg weakness.All patients presented hemifacial spasm,tongue muscle atrophy and fibrillation,amyotrophy and gynaecomastia; creatine kinase level was elevated and endocrine disorder appeared in different degrees.The electromyogram characteristics included widespread neurogenic changes accompanied with/without sensory or motor conduction abnormalities.The forced vital capacity,forced expiratory volume in first second,maximal voluntary ventilation and peak expiratory flow rate were significantly lower,and the residual volume in the KD patients was statistically higher than that in the healthy volunteers (P＜0.05).Respiratory muscle strength (maximum inspiratory pressure and maximum expiratory pressure) decreased in the KD patients.Conclusion KD is a degenerative disease with slow clinical progression which has its own characteristics of inheritance pattern and natural course; the age of onset,repeat number of CAG sequences,pulmonary function and respiratory muscle strength may be valuable for illness evaluation.

Objective To explore the association between diabetes mellitus type Ⅱ (DM) and deglutition function of amyotrophic lateral sclerosis (ALS) patients.Methods Seventy-five ALS patients older than 45 years and admitted to our hospital from August 2008 to November 2015 were selected into this study;67 of them were without DM and 8 were with DM.Water swallow test,amyotrophic lateral sclerosis severity scale-swallow (s-ALSSS),amyotrophic lateral sclerosis functional rating scale (ALSFR-R) and videofluoroscopic swallow study (VFS) were performed to evaluate the deglutition functions of these ALS patients with or without DM,and the results of the two groups were compared.Results (1) Patients with DM had significantly higher s-ALSSS scores,ingurgitation part in ALSFRS-R scores and in parts of VFS scores,such as transportation to pharyngeal,pharyngeal transit,flow into pharyngeal before reflexion,epiglottic vallecula residue,and piriform sinus residue than patients without DM (8.88±1.34 vs.7.54±1.47,3.50±0.54 vs.2.96±0.77,2.88±0.35 vs.2.16±0.69,2.75±0.46 vs.2.09±0.69,2.88±0.35 vs.2.42±0.56,2.88±0.35 vs.2.39±0.58,P＜0.05).(2) Scores of Kubota drinking test,ALSFRS-R,VGF (including oral phase,pharyngeal phase and aspiration degree) in patients without DM were 2.15±1.12,7.18±1.41(1.78±0.69,1.69±0.60 and 3.72±0.65),and those in patients with DM were 1.88±1.34,8.00±0.93(2.13±0.64,2.00±0.53 and 4.00±0.00);no significant differences were noted between the two groups (P＞0.05),but there was a trend showing that DM patients had higher scores in these evaluations.Conclusion As compared with ALS patients without DM,ALS patients with DM get more mildly impaired deglutition function.