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A vaccine is an important method to control schistosomiasis. Molecules related to lung-stage schistosomulum are considered potential vaccine candidates. We previously showed that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and cathepsin L3 (CL3) displayed differential expression in the lung-stage schistosomula of Schistosoma japonicum cocultured with host cells. In the present study, we prepared the two proteins and detected the protective effects of SjGAPDH by immunizing mice with this protein alone and in combination with SjCL3 with or without Freund's adjuvant. Then, we investigated the possible mechanisms underlying S. japonicum infection. The results showed that vaccination of adjuvanted SjGAPDH decreased the worm burden (37.8%) and egg load (38.1%), and the combination of adjuvanted SjGAPDH and SjCL3 further decreased the worm burden (65.6%) and egg load (70.9%) during Schistosoma japonicum infection. However, the immunization of a combination of adjuvant-free SjGAPDH and SjCL3 displayed a lower protective effect (15%) than those of the adjuvanted SjCL3, the adjuvanted SjGAPDH, and a combination of adjuvanted SjGAPDH and SjCL3. Flow cytometric results showed that the frequency of regulatory T cells (Tregs) was lower (P0.05) in the group with adjuvanted SjGAPDH and SjCL3 (2.61%) than the remaining groups. The enzyme-linked immunosorbent assay (ELISA) results indicated that except for the uninfected and infected control groups, the remaining groups displayed a Th1-type shift in immune responses. These results showed the immunization of SjGAPDH resulted in partial protection (approximately 38%); inoculation with a combination of SjCL3 and SjGAPDH in Freund's adjuvant resulted in a high immunoprotective effect (65%) against Schistosoma japonicum infection in mice, which was possibly caused by the reduced percentage of Tregs and a Th1-type shift in immune responses; and SjCL3 has no adjuvant-like effect, dissimilar to SmCL3.

Schistosomiasis is a zoonotic parasitic disease caused by the trematode blood flukes of the genus Schistosoma. The prodigious egg output of females is the main cause of the disease in definitive hosts, while the female worm relies on continuous pairing with the male worm to fuel the growth and maturation of the reproductive organs and egg production. Prohibitin, which contains the functionally interdependent PHB1 and PHB2 subunits in human and some other species, has been proposed to participate in the cell proliferation and apoptosis regulation in mammals. However, little is known about the function of PHB homolog in the growth and reproductive development of schistosomes. Here, we reported the Phb1 gene that was structurally and evolutionarily conserved in Schistosoma japonicum when compared with that of other species from Caenorhabditis elegans to human. Real-time PCR detected that SjPhb1 was highly transcribed in the vitellaria of female worms. SjPhb1 knockdown achieved through the dsRNA-mediated RNAi in vivo resulted in retarded growth, decreased pairing, and fecundity in adult worms, as well as attenuated pathogenicity or virulence of worms to their hosts. Cell proliferation and apoptosis examination found decreased cell proliferation and increased cell apoptosis in SjPhb1 dsRNA-treated worms. Therefore, our study provides the first characterization of S. japonicum PHB1 and reveals its fundamental role in the regulation of growth and development of S. japonicum by specific dsRNA-mediated RNAi in vivo. Our findings prompt for a promising molecular of schistosomes that can be targeted to effectively retard the growth and development of the schistosomes.

Schistosomiasis is still prevalent and seriously endangering the health of people and livestock in many countries. There have been great efforts to develop vaccines against schistosomiasis for prolonged protection in epidemic areas. Molecules from lung-stage schistosomula have been regarded as potential vaccine candidates against schistosomiasis. Our previous work has shown that cathepsin L3 from Schistosoma japonicum (SjCL3) is expressed in lung-stage schistosomula, but its role is not well known. In the present study, we characterized SjCL3 and detected its effect as a possible vaccine in vivo and in vitro. From the results of quantitative PCR (qPCR) and western blot, SjCL3 was present throughout the lifecycle of the worm, and its relative expressed level was higher in the liver eggs and adult worms than other stages. Additionally, immunofluorescence assay showed that SjCL3 was mainly concentrated in the eggshell, alimentary canal, and musculature of worms. Compared with the adjuvant group, the immunization of SjCL3 in mice resulted in a 28.9% decrease in worm burden and a 29.2% reduction in egg number in the host liver. In antibody-dependent cell-mediated cytotoxicity (ADCC) insecticidal experiments in vitro, the existence of SjCL3 could in part suppress adherence between macrophages and worm. The above results indicated that the immunization of SjCL3 could induce limited immune protection against S. japonicum infection in mice, and this protease played a role in breaking the process of ADCC, which was beneficial to the survival of worms.

Objective: To investigate the effects of suspension laparoscopy and pneumoperitoneum laparoscopic surgery on inflammatory response, stress hormone and ovarian function in patients with ovarian cystectomy. Methods: Retrospective analysis of the data of laparoscopic ovarian cyst denudation in our hospital from July 2015 to August 2017, and they were were divided into the control group and the treatment group according to the surgical methods they accepted. The levels of inflammatory factors and stress hormone preoperative and postoperative 24 h, and ovarian function of the menstrual period 3rd day preoperative and postoperative 3 months of the two groups were compared. Results: There was no significant difference in the levels of TNF-α, CRP, COR, NE, E2, LH and FSH between the two groups before the operation. Postoperative 24 h, the levels of TNF-α, CRP, COR and NE in the treatment group were significantly lower than those in the control group, and the level of E2 in the treatment group was higher than that in control group, the levels of LH and FSH in the treatment group were lower than those in control group postoperative 3 months, the difference were statistically significant. Conclusion: Compared with pneumoperitoneum laparoscopic surgery, suspended laparoscopic surgery can effectively reduce the inflammatory response, reduce the level of stress hormone and improve ovarian function in patients undergoing ovarian cystectomy, which has a certain clinical value.

It has been discovered that the development of schistosome is hampered in immunodeficient mice, e.g. nude mice lacking T-lymphocytes and the severe combined immune deficient (SCID) mice lacking both T- and B-lymphocytes. However, it’s still unresolved about the underlying regulatory mechanisms of the retarded growth and development of schistosomes in their immunodeficient definitive host. In this study, therefore, five replicates of male or female Schistosoma japonicum samples with twenty male or female worms in each sample, were collected from SCID mice or BALB/c mice at five weeks post infection and used to perform metabonomic analysis using liquid chromatography tandem mass spectrometry (LC-MS/MS) platform, for elucidating the growth and development regulation of schistosome in their definitive hosts from the metabolomic aspect. Based on the identified 1015 ion features in ESI+ mode and 342 ion features in ESI-mode, multivariate modelling methods including the Principal Component Analysis (PCA), Partial Least Squares Discriminant Analysis (PLS-DA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) identified distinct metabolic profiles that clearly differentiated both male and female worms in SCID mice from those in BALB/c mice, respectively. Common and uniquely perturbed metabolites and their involved metabolic pathways were identified in male and female worms from SCID mice when compared with those from BALB/c mice. The results also revealed that more differential metabolites were found in female worms (one metabolite was up-regulated and forty metabolites were down-regulated) than male worms (nine metabolites were up-regulated and twenty metabolites were down-regulated) between SCID mice and BALB/c mice. The top five increased metabolites of male worms in SCID mice when compared with those in BALB/c mice were PC(22:6/20:1), L-allothreonine, L-serine, glycerophosphocholine and 5-aminoimidazole ribonucleotide. And the top five decreased metabolites of male worms in SCID mice when compared with those in BALB/c mice were PC(16:0/0:0), PAF C-16, PE(18:1/0:0), adenosine and butenoyl PAF. Most of the differential metabolites of female worms in SCID mice had lower levels when compared with the normal female worms in BALB/c mice, except for retinyl ester with a higher level. The top five decreased metabolites of female worms in SCID mice when compared with those in BALB/c mice were adrenic acid, 5-phosphoribosylamine, PC(16:0/0:0), PC(22:6/20:1) and ergothioneine. The involved metabolic pathways of the differential metabolites in male worms between SCID mice and BALB/c mice mainly included taurine and hypotaurine metabolism, glycerophospholipid metabolism, sphingolipid metabolism, arachidonic acid metabolism, alpha-linolenic acid metabolism, etc. The involved metabolic pathways of differential metabolites in female worms included mainly pyrimidine metabolism, sphingolipid metabolism, arachidonic acid metabolism, glycerophospholipid metabolism, tryptophan metabolism, etc. These findings suggested a correlation between the retarded growth and development of schistosome in SCID mice and their perturbed metabolic profiles, which also provided a new insight into the regulation mechanisms of growth and development of S. japonicum worms from the metabolic level, and provided clues for discovery of drugs or vaccines against the parasites and parasitic disease.Author summaryThe growth and development of schistosome has been discovered hampered in the immunodeficient hosts. But it remains unresolved about the molecular mechanisms involved in this. In this study, we tested and compared the metabolic profiles of the male and female Schistosoma japonicum worms collected from SCID mice or BALB/c mice at five weeks post infection using liquid chromatography tandem mass spectrometry (LC-MS/MS) platform. There were 1015 ion features in ESI+ mode and 342 ion features in ESI-mode were identified, and distinct metabolic profiles were found to clearly differentiate both male and female worms in SCID mice from those in BALB/c mice, respectively. The results also found more differential metabolites in female worms than in male worms between SCID mice and BALB/c mice. The enriched metabolic pathways of the differential metabolites in male worms between SCID mice and BALB/c mice included taurine and hypotaurine metabolism, glycerophospholipid metabolism, sphingolipid metabolism, arachidonic acid metabolism, alpha-linolenic acid metabolism, etc. And the enriched metabolic pathways of differential metabolites in female worms included pyrimidine metabolism, sphingolipid metabolism, arachidonic acid metabolism, glycerophospholipid metabolism, tryptophan metabolism, etc. The findings in this study suggested an association between the developmentally stunted schistosome and their perturbed metabolites and metabolic pathways, which provided a new insight into the regulation mechanisms of growth and development of S. japonicum worms from the metabolic level, and clues for discovery of drugs or vaccines against the parasites and disease.

We developed a calcium phosphate cement that could be molded into any desired shape due to its chewing-gum-like consistency after mixing. The powder component of the cement consists of tricalcium phosphate (TCP). The liquid component consists of chitosan, citric acid and glucose solution. In this study, we used four groups of cement to investigate the mechanical properties and biocompatibility of the new biomaterial in vivo. The setting times of the cements were 5-30 min. X-ray diffraction analysis showed that the products were hydroxyapatite (HA) and dicalcium phosphate anhydrous. When the concentration of citric acid was increased, the compressive strength of specimen increased. The animal experiments showed that the material was nontoxic and osteoinductivity.

TNFα played a dominant role in the development and progression of rheumatoid arthritis (RA). Clinical trials proved the efficacies of anti-TNFα agents for curing RA. However, most researchers were concentrating on their abilities of neutralizing TNFα, the potencies of different anti-TNFα agents varied a lot due to the antibody-dependent cell-mediated cytotoxicity (ADCC) or complement dependent cytotoxicity (CDC). For better understanding and differentiating the potentiality of various candidate anti-TNF reagents at the stage of new drug research and development, present study established a cell model expressing the transmembrane TNFα for usage in in vitro ADCC or CDC assay, meanwhile, the assay protocol described here could provide guidelines for screening macromolecular antibody drugs. A stable cell subline bearing transmembrane TNFα was first established by conventional transfection method, the expression of transmembrane TNFα was approved by flow cytometer, and the performance of the stable subline in ADCC and CDC assay was evaluated, using human peripheral blood mononuclear cells as effector cells, and Adalimumab as the anti-TNFα reagent. The stable cell subline demonstrated high level of surface expression of transmembrane TNFα, and Adalimumab exerted both ADCC and CDC effects on this cell model. In conclusion, the stable cell line we established in present research could be used in ADCC or CDC assay for screening antibody drugs, which would provide in-depth understanding of the potencies of candidate antibody drugs in addition to the traditional TNFα neutralizing assay.

Previously, the application of cisplatin in chemotherapy was limited due to the significant side effects on normal cell growth. In the present study, the concomitant application of emodin with cisplatin was demonstrated to ameliorate cisplatin-induced oxidative stress and markedly suppress tumor cell proliferation for the first time. Human osteosarcoma MG-63 cells were treated with cisplatin alone or in combination with emodin. The cell viability was determined by MTS assays and the augmentation of reactive oxygen species were determined by fluorogenic probes; in addition, a stable MG-63 subline bearing antioxidant response element (ARE)-driven luciferase expression was developed to monitor the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2)-ARE signaling pathway. The results indicated that cisplatin or emodin may inhibit MG-63 cell proliferation in a time- or dose-dependent manner, respectively. Concomitant treatment with cisplatin and emodin demonstrated synergic anti-tumor effects. Cisplatin augmented reactive oxygen species in the MG-63 cells, followed by the translocation of Nrf2 from the cytoplasm into the nucleus, which triggered ARE-driven luciferase expression. The addition of emodin diminished the previously described phenomenon, resulting in decreased ROS augmentation, translocation of Nrf2 and ARE-driven luciferase activity. In conclusion, emodin could ameliorate cisplatin-induced oxidative stress and protect the cells from oxidative stress-induced damage. The findings of the present study provide a novel strategy for the treatment of osteosarcoma using emodin and cisplatin.

Early serous carcinoma in fallopian tube or serous tubal intraepithelial carcinoma (STIC), an early lesion limited to the epithelium of the fallopian tube and firstly identified from specimen obtained by prophylactic salpingo-oophorectomy, has provided insight into pelvic high grade serous carcinoma (HGSC). Increasing evidence indicates that STIC is a likely precursor for HGSC and several studies have focused on this lesion and its clinical significance. This review addresses recent advances in recognizing STIC and its correlation with HGSC and ovarian carcinogenesis. It also describes evidence regarding the fallopian tube as a source of some HGSCs, the protocol for optimizing histological evaluation of the tubes, the spectrum of tubal lesions from benign to noninvasive carcinoma, changes in diagnostic criteria from purely morphologic characteristics to a combination of morphologic features and molecular biomarkers, and new studies about potential biomarkers. However, the direct evidence regarding STIC as the precursor of HGSC is still tantalizing due to other possibilities that may also explain the origin of pelvic HGSC. Further molecular genetic studies are required to address this important question.

To investigate the effects of miR-135a on HOXA10 expression, proliferation and apoptosis of SKOV3 cells.(1) Through computer-aided algorithms,the predicted target gene of miR-135a (HOXA10)were determined. (2) miR-135a mimics, miR-135a inhibitor and negative control were transfected into SKOV3 cells, respectively.Reverse transcription (RT)-PCR, western blot analysis were used to examine the expression levels of HOXA10 at different times (24, 48 and 72 hours). (3) A luciferase reporter assay was used to confirm the direct regulation between miR-135a and HOXA10. (4) SKOV3 cells proliferation at different times (24,48 and 72 hours) was detected by methyl thiazolyl tetrazolium (MTT) assay [quantified by absorbance(A)]. Western blot was used to examine the expression of apoptosis-associated protein bcl-2, bax and caspase-3 in SKOV3 cells after 48 hours transfection.(1) HOXA10 was predicted to be the target gene of miR-135a by computer-aided algorithms. (2) RT-PCR shown that HOXA10 mRNA levels were decreased over time (24, 48 and 72 hours) after miR-135a mimics transfection in SKOV3 cells (0.94 ± 0.04 vs 0.78 ± 0.03 vs 0.70 ± 0.03, P0.05). While, the expression of HOXA10 mRNA was increased over time after miR-135a inhibitor transfection (1.14 ± 0.05 vs 1.16 ± 0.03 vs 2.60 ± 0.08,P0.05). After transfected with miR-135a mimics or miR-135a inhibitor over 48 and 72 hours, the HOXA10 expression levels in SKOV3 cells were significantly lower or higher than each control group, respectively (all P0.01). Western blot analysis of HOXA10 expression in SKOV3 cells confirmed the results of RT-PCR detected. (3) After cotransfection of miR-135a plasmid and pMIR-REPORT luciferase plasmid containing HOXA10, luciferase reporter assays showed that the luciferase activity reduced by 67.8% (P0.01). (4) MTT showed that SKOV3 cells growth after miR-135a mimics transfection for 48 and 72 hours were significantly lower than those in control group (0.38 ± 0.03 vs 0.52 ± 0.05, 0.67 ± 0.05 vs 0.75 ± 0.06;respectively,all P0.05).While, SKOV3 cells transfected with miR-135a inhibitor for 72 hours grew significantly faster than that in control group (0.95 ± 0.05 vs 0.75 ± 0.06, P0.01). After miR-135a mimics transfection, the level of bcl-2 protein was significantly lower than that in control group (0.28 ± 0.06 vs 0.76 ± 0.09,P0.01). The activity of caspase-3 was significantly higher than that in control group (115.0 ± 2.4 vs 95.4 ± 2.1, P0.01). While, there was no statistical difference of bax expression (P = 0.142). However, after miR-135a inhibitor transfection, the expression level of bcl-2 protein was significantly higher than that in control group (0.92 ± 0.03 vs 0.76 ± 0.09, P = 0.037) and the activity of caspase-3 was significantly lower than that in control group (59.5 ± 4.1 vs 95.4 ± 2.1, P0.01). There was also no statistical difference of bax expression (P = 0.066).miR-135a may play an important role in cell proliferation and apoptosis of ovarian cancer cells by regulating HOXA10 and its downstream pathways.

To prepare a goat model of tibial bone hole defect suitable for studies of bone defect repair using tissue-engineered injectable bone materials.A circular hole bone defect 1.2 cm in diameter was induced below the tibial medial plateau of the goat. X-ray, histological inspection, and image analysis were carried out to evaluate the validity of the model in simulating limb bone defect for the study of tissue-engineered injectable bone materials.At 4 and 8 weeks after the operation, neither X-ray nor histological examination showed obvious bone tissues in the bone defect. Image analysis showed a area of new bone tissue formation of (8.79 - or + 3.63)% in the total defect area at 4 weeks, which increased to (15.41 - or + 4.21)% at 8 weeks.The goat model of tibial bone hole defect established in this study is suitable for studying the ability of injectable bone materials for repairing limb bone defect, and offers a simple and reliable means to simulate the local condition of bone regeneration and mechanical environment of bone defect in the limbs.