Background: Peste des petits ruminants (PPR), caused by the PPR virus (PPRV), is an acute and fatal contagious disease that mainly infects goats, sheep, and other artiodactyls.Peripheral blood mononuclear cells (PBMCs) are considered the primary innate immune cells. Objectives: PBMCs derived from goats were infected with PPRV and analyzed to detect the relationship between PPRV replication and apoptosis or the inflammatory response. Methods: Quantitative real-time polymerase chain reaction was used to identify PPRV replication and cytokines expression. Flow cytometry was conducted to detect apoptosis and the differentiation of CD4+ and CD8+T cells after PPRV infection. Results: PPRV stimulated the differentiation of CD4+ and CD8+ T cells. In addition, PPRV induced apoptosis in goat PBMCs. Furthermore, apoptosis and the inflammatory response induced by PPRV could be suppressed by Z-VAD-FMK and Z-YVAD-FMK, respectively.Moreover, the virus titer of PPRV was attenuated by inhibiting caspase-1-dependent apoptosis and inflammation. Conclusions This study showed that apoptosis and the inflammatory response play an essential role in PPR viral replication in vitro, providing a new mechanism related to the cell host response.

Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases.The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions The spbELISA has practical applications in assessing the vaccination status of large pig herds.

Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Singledomain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Singledomain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

Objective To investigate the possible mechanism of coix seed in inducing apoptosis of gastric cancer cells (SGC-7901). Methods Different concentrations of coix seed oil (2, 4, 8 mg/mL) were applied to SGC-7901 cells. MTT assay was used to detect the effect of drugs on cell proliferation, and flow cytometry for drug-induced cell apoptosis, scratch test for cell migration inhibited by coix seed oil, Transwell chamber for drug-inhibited cell invasion, and Western blot for expression of related proteins PRMT5, PI3 K and AKT. Results The results of MTT showed that 2, 4 mg/mL of coix seed oil could significantly inhibit the proliferation of SGC-7901 cells, and the inhibition rate of 2 mg/mL was (30. 02 ± 1. 56) ％, which showed significant difference compared to the control group (P ＜ 0. 01). The results of flow cytometry showed that the apoptosis rates of coix seed oil cells at concentrations of 2 and 4 mg/mL were (16. 25 ± 2. 54) ％, (12. 60 ± 1. 12) ％, respectively, and which showed a significant difference compared with (2. 0 ± 1. 22) ％ in the control group (P ＜ 0. 01). The results of cell scratch test showed that the migration of SGC-7901 cells treated with 2, 4 mg/mL of coix seed oil had significant difference when compared to control group (P ＜0. 01). The results of invasion experiments showed that 2, 4 mg/mL of coix seed oil could significantly inhibit cell invasion, and the number of cell invasion was (134. 00 ± 2. 86), (167. 00 ±0. 99), respectively, which showed significant difference compared to (268. 00 ± 2. 05) in the control group (P ＜ 0. 01). The migration number in 8 mg/mL coix seed oil group was (167 ± 0. 99), a significant difference was observed when compared to the control group (P ＜ 0. 05). Western blot analysis showed that different concentrations of coix seed oil could significantly down-regulate the expression of PRMT5, PI3 K and AKT in SGC-7901 cells. Conclusion Coix seed oil can significantly inhibit the proliferation, migration and invasion of gastric cancer SGC-7901 cells, its possible mechanism is to down-regulate the signal pathway of PRMT5-PI3 K/AKT to inhibit the activation of various anti-apoptotic molecules, induce apoptosis and suppress invasion and metastasis of gastric cancer SGC-7901 cells by down-regulating the PRMT5-PI3 K/AKT signaling pathway.

Objective To investigate the possible mechanism of coix seed in inducing apoptosis of gastric cancer cells (SGC-7901). Methods Different concentrations of coix seed oil (2, 4, 8 mg/mL) were applied to SGC-7901 cells. MTT assay was used to detect the effect of drugs on cell proliferation, and flow cytometry for drug-induced cell apoptosis, scratch test for cell migration inhibited by coix seed oil, Transwell chamber for drug-inhibited cell invasion, and Western blot for expression of related proteins PRMT5, PI3 K and AKT. Results The results of MTT showed that 2, 4 mg/mL of coix seed oil could significantly inhibit the proliferation of SGC-7901 cells, and the inhibition rate of 2 mg/mL was (30. 02 ± 1. 56) ％, which showed significant difference compared to the control group (P ＜ 0. 01). The results of flow cytometry showed that the apoptosis rates of coix seed oil cells at concentrations of 2 and 4 mg/mL were (16. 25 ± 2. 54) ％, (12. 60 ± 1. 12) ％, respectively, and which showed a significant difference compared with (2. 0 ± 1. 22) ％ in the control group (P ＜ 0. 01). The results of cell scratch test showed that the migration of SGC-7901 cells treated with 2, 4 mg/mL of coix seed oil had significant difference when compared to control group (P ＜0. 01). The results of invasion experiments showed that 2, 4 mg/mL of coix seed oil could significantly inhibit cell invasion, and the number of cell invasion was (134. 00 ± 2. 86), (167. 00 ±0. 99), respectively, which showed significant difference compared to (268. 00 ± 2. 05) in the control group (P ＜ 0. 01). The migration number in 8 mg/mL coix seed oil group was (167 ± 0. 99), a significant difference was observed when compared to the control group (P ＜ 0. 05). Western blot analysis showed that different concentrations of coix seed oil could significantly down-regulate the expression of PRMT5, PI3 K and AKT in SGC-7901 cells. Conclusion Coix seed oil can significantly inhibit the proliferation, migration and invasion of gastric cancer SGC-7901 cells, its possible mechanism is to down-regulate the signal pathway of PRMT5-PI3 K/AKT to inhibit the activation of various anti-apoptotic molecules, induce apoptosis and suppress invasion and metastasis of gastric cancer SGC-7901 cells by down-regulating the PRMT5-PI3 K/AKT signaling pathway.

Objective To identify the influence of chemotherapy-induced serum alkaline phosphatase (ALP) elevation on the tumor-free survival (TFS) in patients of gastric carcinoma after radical gastrectomy.Methods The clinical data of 189 gastric carcinoma patients receiving radical surgery and postoperative adjuvant chemotherapy between Jan,2010 and Dec,2015 were reviewed and statistically analyzed.Results The TFS of patients with serum ALP elevation was obviously inferior than those without ALP elevation (x2 =5.717,P =0.017),serum ALP elevation was an independent risk factor influencing patients' TFS (HR =2.178,P =0.032),the degree of serum ALP elevation was associated with patients' TFS (x2 =4.627,P =0.031).Conclusion Serum ALP elevation during postoperative chemotherapy indicates the increases of recurrence or metastasis rate of gastric cancer patients after radical gastrectomy.

OBJECTIVETo compare the effect and differences sex the influence of hormone levels of perimenopau-sal syndrome patients between manual acupuncture and electroacupuncture (EA).

METHODSA total of 50 cases with perimenopausal syndrome were randomly assigned into an manual acupuncture group (27 cases) and an EA group (23 cases), and 1 case dropped in the EA group. The acupoints in the two groups were Guanyuan (CV 4), Zigong (EX-CA 1), Tianshu (ST 25), and Sanyinjiao (SP 6). Acupuncture with 3-time small and even manipulation of lifting, thrusting and twirling was used in the acupuncture group, once 10 min. EA with sparse-dense wave and 10 Hz/50 Hz was applied in the EA group for 30 min. The treatments in the two groups were for continuous 8 weeks (24 times in total), once the other day, 3 times a week. The scores of 24-hour hot flashes even, menopausal rating scale (MRS) and menopause-specific quality of life questionnaire (MENQOL) were recorded before treatment and after 4-week and 8-week treatment, as well as 12 and 24 weeks after treatment. Serum sex hormone levels were tested before and after 8-week treatment as well as 12 weeks after treatment, including serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estracliol (E).

RESULTSCompared with those before treatment, the 24-hour hot flashes even score, MRS and MENQOL scores were significantly lower after 4-week and 8-week treatments, 12 and 24 weeks after treatment (all<0.05). All the above scores after 8-week treatment were lower than those after 4-week treatment (all<0.05); and the scores 12 and 24 weeks after treatment were lower than those after 4-week and 8-week treatments (all<0.05); all the scores after treatment were not significantly different at any time between the two groups (all>0.05). Compared with those before treatment, serum FSH and Eapparently improved in the two groups after 8-week treatment and 12 weeks after treatment (all<0.05). LH levels did not significantly change in the two groups (all>0.05). All the serum sex hormone levels showed no significant difference between the two groups (all>0.05).

CONCLUSIONSBoth acupuncture and EA can improve perimenopausal symptoms and serum sex hormone. The effects are similar.

Objective To investigate the diagnostic value of multiparametric MRI in early prostate cancer(PCa) based on PI-RADS version 2.Methods 27 surgically-proved early PCa patients were collected in this retrospective study.T2WI,DWI and DCE were evaluated by two blinded radiologists.By 12 sub-region classification method the possibility of the presence of cancer at each sub-region was scored according to the PI-RADS V2.The receiver operating characteristic (ROC) curve was used to analyze the diagnosic efficacy of the following 4 protocols:T2WI alone(protocol 1),T2WI+DWI(protocol 2),T2WI+DCE(protocol 3),T2WI+DWI+DCE(protocol 4).The sensitivity,specificity and accuracy for each protocol were calculated.The average scores of cancerous sub-regions and non-cancerous sub-regions were calculated and the independent sample t test was used to compare the four protocols.Results 324 sub-regions were analyzed in 27 early PCa patients and then divided into 119 cancerous sub-regions and 205 non-cancerous sub-regions,including 64 peripheral zone cancerous sub-regions and transition zone cancerous sub-regions.In protocol 1-4, the average scores of cancerous sub-regions in orderwere 3.13±1.19,3.27±1.15,3.28±1.23, 3.33±1.16,respectively.Non-cancerous sub-regions's scores in order were 1.98±0.90,1.91±0.91, 2.03±0.99,1.94±0.96 respectively and there were significant differences among each protocol (P<0.05).The area under the ROC curve of the 4 protocols for region-based analysis were displayed in descending order: protocol 4 (0.819), protocol 2 (0.810), protocol 3 (0.772), protocol 1 (0.765) and there were no significant differences between any two protocols (P>0.05).In four protocols, the sensitivity in order were 45.40%, 56.30%, 59.70%, 61.34%, while the specificity in order were 95.10%, 96.10%, 89.80%, 96.60%, and the accuracy in order were 76.85%, 81.48%, 78.70%, 83.65%.Conclusion Multiparametric MRI can improve the diagnostic accuracy for the detection of early PCa, and T2WI+DWI+DCE is with the highest value.The PI-RADS V2 system is a better semi quantitative method for evaluation of early PCa.

Objective To evaluate the value of T2WI scores for diagnosis and differential diagnosis of transition zone prostate cancer(PCa).Methods T2WI of 43 cases of transition zone PCa and 91 cases of benign prostatic hyperplasia(BPH) were analyzed retrospectively.Transition zone T2WI signs were divided into the main signs and the secondary signs, which were given different scores and were evaluated separately by receiver operating characteristic (ROC) curve for their diagnostic value.Results In a total of 11 scores sections (-1-10), as the increase of scores, the sensitivity of transition zone PCa was decreased while the specificity and positive predictive value were both increased.According to the ROC curve, when the critical value of scores≥4.5, the sensitivity, specificity, negative predictive value, and the accuracy were 81%,73.3%,90.9%,70.4% respectively.When the scores ≥8.5,the specificity and positive predictive value were both 100%.When the scores ≥0.5 and ≥1.5,the negative predictive value were 100%.Conclusion T2WI scores can quantitatively analyze the transition zone PCa,which has an important value to improve the clinical diagnosis and guide treatment.