Objective To explore the effects of maternal hypercholanemia on the myocardium changes in rat fetus. Methods Thirty clean SD female rats were equally randomized to three groups after mating successfully.From the 13th to 20th day of gestation,group A and B were injected injected with sodium chloride(NS) as control.Total bile acid(TBA) and cardiac troponin I(cTnI) were measured in the maternal and fetal serum on the 21st day when all rats were killed.Fetal cardiac muscle cells were also collected for examination with light microscope and electronic microscope.Results (1)TBA in maternal and fetal serum were(22.32±8.12)μmol/L and(28.84±8.06) μmol/L,respectively in group A,(9.77±3.56)/μmol/L and(9.34±3.54) μmol/L in group B,and (3.60±1.78) μmol/L and(3.95±1.19) μmol/L in group C.Significant differences were found among groups(P<0.01).(2)Fetal death rates were significantly different among the three groups (P<0.05),with 30.11%,16.85%,and 7.05%,respectively.(3)Fetal cTnl were also found significant difference among groups(P<0.01),with(19.98±7.75)ng/ml,(11.41±3.64)ng/ml and(4.38±1.19)ng/ml,respectively.(4)The integrated scores of fetal necrosis area were significantly different in three groups(P<0.05),with 1.92±0.43,1.36±0.37 and 0.44±0.12,respectively.(5)Under electronic microscope,the number density of mitochondria in group A was lower than that in group C(P<0.05)while the average volume of mitochondria was larger in group A (P<0.05).The average volume of mitoehondria in group B was larger than that in group C(P<0.05) while no difference was found with regard to the number density between the two groups.The number density and average volume of myofibril in group A were lower than those in group C(P<0.05).The number density of myofibril in group B was higher than that in group C(P<0.05) while no difference was found with the average volume.(6)Positive correlations were found in maternal TBA,fetal TBA,fetal cTnI and the integrate of fetal necrosis area when comparing every two of the above factors. Conclusions Fetal myocardium is impaired obviously in hypercholanemia rats.The serum level of TBA and cTnI in fetal rats are positively correlated with each other.

Objective To study epidemiological characteristics of thelazjasis, in Zunyi and surrounding areas, morphology and 18S rRNA gene sequence of Thelazia callipaeda. Methods The Thelazia callipaeda was collected from several hospitals, including affiliated Hospital of Zunyi Medical College and Zunyi Aerospace Hospital, in Zunyi region since 2011 to 2015. Clinical data of infected human thelaziasis, including the patients' gender, age, residence and pets such as cats or dogs, were analyzed to find out the factors influencing the incidence. Morphology characteristics of female and male Thelazia callipaeda were observed under microscope. 18S rRNA gene of Zunyi Thelazia callipaeda was amplified by PCR, and the evolutional relationship was analyzed through the software MEGA 7.01 based on neighbour-joining (NJ) method. Homology was campared with 18S rRNA gene from GenBank in National Center of Biotechnology. Results Totally 25 cases had been reported during the study, of which 22 cases had more details. Based on the cases, we found the thelaziasis was increasing year by year. For instance, 2 cases (9.1%) were reported in 2011, 1 case (4.5%) in 2012, 3 cases in 2013 (13.6%), 10 cases (45.5%) in 2014 and 6 cases (27.3%) in 2015 . During the five years , totally 15 cases were treated between August and November, when the human thelaziasis was in typical epidemic peaks. We analyzed characteristics of the total cases reported to date. Most of the cases occurred in rural areas (20 cases). Majority of patients lived in rural region. And most cases were between 30 to 60 years old, indicating no age limit, especially, there were two cases who were at the ages of 8.5 months and 77 years old, respectively. Moreover, more women suffered from the disease than that in men, of which, the case number was 16 in women and 6 in men, and there were 7 cases who had cats or dogs at home. Under light microscope, the edge of Thelazia callipaeda had serrated cuticle with transverse striations. And male worm had a sharp peak at the tail end of Thelazia callipaeda, which cured to the ventrite and had two copulatory spicules, long one and short one, respectively. While female worm had a blunt tail, containing numerous eggs and rounded first-stage larvaes in a single line in the distal area of the uterus, near the vulva. Sequences of Thelazia callipaeda 18S rRNA gene from Zunyi and Oita Japan (AB538282.1) were showed homology of 100%, and they were clustered in a same branch of Phylogenetic tree. Conclusions Human thelaziasis cases in Zunyi region are increasing each year, and most of the cases have occurred in rural areas. 18S rRNA gene has a high homology with sequence AB538282.1 in Thelazia callipaeda. Combining clinical data, analysis of epidemiological characteristics and the characteristics of 18S rRNA should be good for specie identification and epidemiological analysis.

The serum lipid and lipoprotien cholesterol levels of 1974 healthy subjects from newborn to 75 years of age and 51 patients with coronary heart disease (CHD) and cerebrovascular disease(CVD) were studied in Wuhan district. The serum lipid and lipoprotein cholseterol concentration varies with age. But no differences were found between male and female cord blood. In both sexes the serum T-C, LDL-C, HDL-C, T-C/HDL-C and LDL-C/ HDL-C ratio increased with the increasing of age. The serum HDL-C level has significant difference between male and female (P

Objective To investigate the relationship of alcohol exposure during pregnancy and non-compaction cardiomyopathy (NCC) in offspring mice.Methods Pregnant mice of ED3.5-ED18.5 were given 56％ alcohol by gavage at a dose of 5 mL/kg.The ED19.5 mice were sacrificed,and the heart of the fetal mice was harvested.Transmission electron microscopy (filaments,mitochondria and sarcoplasmic reticulum) and HE staining were used to verify the changes of structure and ultrastructure of the obtained myocardial tissues.Echocardiography was used to evaluate the cardiac function and ventricular myometrium of the offspring mice after growing up.Results Alcohol exposure during pregnancy caused the disorganized and dissolved myofilaments in the fetal mice.Some offspring mice (31.25％,5/16) had NCC.The ratio of non-compacted myocardium to compact myocardium at the end of systole (N/C) was 2.49 ± 0.6 in the offspring mice of the alcohol exposure group,significantly higher than that in the control offspring mice (0.62 ± 0.23,t =10.397,P =0.000).The volume of heart was decreased in the offspring mice of the alcohol exposure group while the left ventricule was enlarged.Echocardiography showed cardiac dysfunction and thickened ventricular septal/left ventricular posterior walls in the grown-up mice of the exposure group.Conclusion Large dose of alcohol exposure during pregnancy cause trabeculations and non-compaction in ventricular myocardium,and it might be one of causers for NCC in the offspring.

Objective To evaluate the feasibility of negative pressure suction enteroscopy for intestinal examination.Methods The negative pressure suction enteroscopy was first applied to examine 20 detached small intestines from pigs to explore a suitable work pressure and to detect possible intestinal damages.The procedure was then applied in living pigs to evaluate the feasibility and to observe possible adverse reactions in a 2-week follow-up.Resnlts The absolute working pressure of the negative pressure suction enteroscopy should be higher than 0.06 Mpa.The procedures were successfully completed in all 20 pig small intestines in 40 + 20 min.There was no tissue damage in the intestinal wall according to histological findings.Local glide occurred in living pig examination at 0.06 Mpa,and it disappeared when the absolute value was higher than 1.00 Mpa.The procedure was completed in 20 living pigs in 60 ± 20 min.Histological findings showed complete tissue structure,clear villi and complete submucous and muscular tissue.All pigs could take food at recovery from anaesthesia.18 had bowel movement on the 2nd day after the operation,and the 2 others recovered on the 3rd day.No abdominal distension,hematochezia,or constipation was observed in 2-week follow-up.Food intake,activity,sleep,urination and defecation were all normal.Conclusion The negative pressure suction enteroscopy is effective and safe for small intestine examination,and is of good prospect.

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 effect of parvovirus B19 (VB19) infection on pediatric leukemia patients. Methods The pediatric leukemia patients were enrolled in the study in the Children's Hospital of Soochow University. Expression levels of VB19-DNA-PCR were detected using the polymerase chain reaction. Positive patients would be monitored and treated by conventional treatment as well until VB19 gene became negative. The data was compared according to the VB19 clearance time, clinical symptoms and blood counts to evaluate the effect. Results In the 3009 samples from 824 pediatric leukemia patients, there were 36 samples (1.2%) from 12 cases (1.5%) of pediatric leukemia paients with positive VB19 infection. Among the positive patients, 11 cases (1.9%) were from 582 with acute lymphoblastic leukemia (ALL) patients and 1 (0.45%) was from 212 with acute myeloid leukemia (AML). According to the treatment stage, 3 cases were in initially diagnosed period, 2 cases in early stage of consolidation chemotherapy, 4 cases in delayed enhanced chemotherapy period, and 3 cases in maintenance chemotherapy period. According to the treatment response, 4 cases were in continuous treatment, 2 cases were sensitive to treatment, and 3 cases were drug resistant. In the drug resistance group, 2 cases developed into the pure red cell aplastic anemia (PRCA). After treatment, one was recovered from PRCA with VB19 cleared, the other one remained PRCA with continuously positive VB19. Conclusions More VB19 virus infection in pediatric ALL happened in delayed enhanced chemotherapy period. The persistent presence of VB19 infection on pediatric leukemia patients is closely related with PRCA.

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.