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 dosimetric effect of using a split of the gross tumor volume (GTV) delineation approach during intensity-modulated radiotherapy (IMRT) treatment planning for advanced T-stage nasopharyngeal carcinoma (NPC) with prevertebral space involvement (PSI).Methods Twenty NPC patients with T3-T4 tumors with prevertebral space involvement were studied.Two kinds of plans were generated based on the same protocol.The first plan was GTV-split consisting of organ at risk (including spine cord and brain stem) overlapping and non-overlapping subsegments.The subsegments were assigned independent dose constraint.The second plan was done based on a standard treatment planning protocol.The two plans were compared with respect to target coverage and organ at risk (OAR) sparing,and target conformity index.Results Two plans both met requirement of dose coverage of target and constraints on normal tissue.Regarding the planning gross target volume (PGTV),the doses of ＜ 93％ volume of plan 1 and plan 2 were [(0.63 ± 0.21)％,and (0.68 ± 0.10)％,P =0.049];the doses of ＞ 110％ volume of plan 1 and plan 2 were [(11.59 ± 7.Ol) ％,and (12.71 ± 6.03) ％,P =0.041];the doses of ＞115％ volume of plan 1 and plan 2 were [(2.05 ± 0.80)％,and (2.76 ± 1.14)％,P =0.049];about OARs,the maximal dose of spine cord for plan 1 and plan 2 were (41.00 ± 4.52) Gy and (45.00 ± 3.25) Gy (P =0.048);and the target coverage and target conformity index of plan 1 were better than those of plan 2,the p-values were 0.063,and 0.135,respectively.The difference between two plans was statistically significant (P ＜ 0.05).Conclusions A split-GTV delineation,independent dose constraint,and multi-course IMRT for advanced T-stage nasopharyngeal carcinoma with prevertebral space involvement optimized IMRT treatment planning,improved target coverage and target conformity index,and decreased the maximal dose for the spine cord and brain stem.It is feasible and has advantage to decrease the dose to OARs.The approach deserves to be further studied.

Objective To evaluate the effects of Taoist cognitive psychotherapy on depression of aged patients with cerebral stroke hemiplegia convalescence. Methods Seventy-seven hemiplegia convalescence patients with depression were divided into general treatment group (38 patients, received general back-up psychology therapy) and combined treatment group (39 patients, received general back-up psychology therapy and Taoist cognitive psychotherapy) by random digits table. All patients were treated for 8 weeks and followed up for 6 months. Two groups were evaluated with HAMD and SS-QOL before treatment and at the end of the 2 weeks,4 weeks, 8 weeks and 6 months after treatment. The results were analyzed with statistics. Results In general treatment group, the HAMD scores were gradually decreased, and the HAMD scores of patients after 8 weeks' treatment were significantly lower than those before treatment[(22.35 ± 4.69)scores vs. (29.62 ± 5.95 ) scores,t = 6.425 ,P ＜ 0.01]. At the end of 6 months after treatment, the scores increased [(24.48 ± 4.12 ) scores vs. (22.35 ± 4.69 ) scores, t = 2.014, P ＜ 0.05], but they were lower than those before treatment(t = 4.836, P ＜ 0.01 ). At the end of 6 months after treatment, the SS-QOL scores were lower than those after 8 weeks' treatment (t =2.543,P ＜0.05),but they were higher than those before treatment (t = 2.790,P ＜ 0.05 ). In combined treatment group, the HAMD scores decreased continuously,and the scores after 8 weeks' treatment [(20.08 ± 4.60) scores] and 6 months' treatment [( 15.21 ± 3.42)scores] were significantly lower than those before treatment [( 30.14 ± 4.92 ) scores] (t = 8.341,15.443, P ＜0.01). Meanwhile,the HAMD scores after 6 months'treatment were significantly Iower than those after 8 weeks' treatment (t =4.724,P ＜ 0.01 ). The SS-QOL scores after 8 weeks' treatment [( 117.56 ± 26.22)scores] and 6 months' treatment [(126.57 ±21.82) scores] were significantly higher than those before treatment[(86.54 ± 23.90) scores] (t = 6.716,8.916,P ＜ 0.01 ) ,and there was significantly difference(t=2.378,P ＜ 0.05). The HAMD scores of combined treatment group after 8 weeks' and 6 months' treatment were significantly lower than those of general treatment group at the same time(t = 2.118, P ＜ 0.05 ;t = 8.405,P＜ 0.01 ) ,and SS-QOL scores were significantly higher than those of general treatment group at the same time (t = 3.123,P ＜ 0.05 ;t = 6.580,P ＜ 0.01 ). Conclusions General back-up psychology therapy combined with Taoist cognitive psychotherapy can improve depression and life quality of cerebral stroke hemiplegia convalescence in aged patients. The effects of Taoist cognitive psychotherapy is slower, but it is more beneficial in the long time.

Objective In recent years,driven by our country's policy of strengthening independent innovation capabilities,there is dramatic increasing of the Patent application volume in Guizhou Province.This article analyzes the patent application and authorization of Guizhou medical research institutes,explored its existing problems and proposed countermeasures.Methods In this paper,the author searched the patent data of Guizhou Medical Scientific Research Institutions,including medical universities and their affiliated hospitals,medical colleges,and people's hospitals at all levels,through the Baiten Patent Data Retrieval Website.Data about the number of patent applications,types of applications,technical composition,legal status,legal events were analyzed to understand the existing problems,and proposed further solutions.Results At present,the medical scientific research units in Guizhou Province still have many problems in patent applications,such as low quality,Pay attention to the application but neglect the maintenance,low patent transactions,lack of patent knowledge for scientific researchers,and low patent consulting services.Conclusions These problems can be solved by improving the intellectual property management department,strengthening information literacy education,running a new mode of school-enterprise cooperation,and adding patent service organizations.

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 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.

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 investigate the significance of aldehyde dehydrogenase 1 (ALDH1)expression in appraising the prognosis of nasopharyngeal squamous carcinoma (NPC).Methods The expression of ALDH1 protein was detected by immunohistochemistry in human specimens obtained from 120 NPC patients without the history of radiotherapy or chemothetapy.Results A relatively high expression of ALDH1 were observed in 40.0 ％ (48/120) patients of NPC,the expression of ALDH1 in the edge of the cancer nests and stroma,particularly spindle cancer cells were strongly positive.The positive expression of ALDH1 was closely associated with patients' nasopharyngeal tumor size (P =0.011),lymphatic metastasis (P =0.005) and clinic stage (P =0.001),but not associated with their gender and age (both P ＞ 0.05).The Kaplan-Merier and Cox regression analysis indicated that ALDH 1 closely correlated with clinical progrosis of the patients.It showed that ALDH1 was an independent risk factor which may affect the prognosis of NPC patients.Conclusion The expression of ALDH1 protein closely correlate with clinic outcome of NPC,suggesting that ALDH1 is a risk factor for clinic progrosis of NPC patients.

Objective To investigate the clinical application and manifestation of dynamic contrastenhanced MRI (DCE-MRI) in differentiating true progession from pseudoprogression in patients with gliobastomas.Methods Twenty five glioma patients were treated with postoperative concurrent chemoradiotherapy and enrolled in this study.All patients were underwent DCE-MRI using a 1.5T scanner.Fifteen patients were confimmed by secondary pathology or clinical and imaging follow-up of patients with gliomas true progession (TP),10 patients were pseudoprogress (PP).Nonparametric Mann-Whitney test was used to compare perfusion parameters between two groups (TP and PP),were used for receiver operating characteristic (ROC) curve analysis to clear if these parameters can be the indicators to differentiate true progession from pseudoprogression.Results Ktrans (volume transfer constant),Ve (fractional volume of extravascular extracellular) values between TP and PP glioma groups were statistically significant,K and Ve values were significantly higher in the TP group than in the PP group (P ＜ 0.05).The areas under the ROC curve are 0.990 and 0.847,respectively.Kep (efflux rate constant) value,Vp (fractional volume of plasma) value in the identification of glioma TP group and PP group was not statistically significant (P ＞ 0.05).Conclusions DCE-MRI can be used to identify glioma TP and PP,Ktrans value and Ve value have clinical significance.

stinguish active tuberculosis and healthy cases with tuberculosis exposure history according SFC count.