Reading Ability in Patients With Tuberous Sclerosis Complex: Results of Chinese Character Reading and Reading Comprehension Tests.

Background: Most tuberous sclerosis complex (TSC) patients have neurological disorders and are at high risk of academic difficulties. Among academic skills, reading ability is the most important academic skill. The study applied the Chinese character fluency test to measure the word recognition and reading comprehension of TSC children to observe whether they have the characteristics of reading disability, as an indicator of the spectrum of reading ability in TSC patients. Methods: The patients were assessed using the Chinese character fluency test and reading comprehension test to explore the differences in reading ability in terms of gender, age, epilepsy history, genotype, and intelligence level. Results: Of the 27 patients, the assessment of reading accuracy showed statistical differences between intellectual level > 80, PR (p = 0.024), and pass numbers (p = 0.018). For the fluency assessment, there was a difference between different intellectual level (p = 0.050). In the reading comprehension test, there was differences for intellectual level in positivity (p = 0.07) and pass numbers (p = 0.06). Conclusion: The Chinese character fluency and reading comprehension test measure the word recognition and reading comprehension and the spectrum of reading ability in TSC patients. All individuals with TSC, especially those with below average of intellectual ability, should be considered for potential academic difficulties.

Continuous low-dose everolimus shrinkage tuberous sclerosis complex-associated renal angiomyolipoma: a 48-month follow-up study.

Tuberous sclerosis complex (TSC) is a rare disease that causes multisystem benign neoplasm, induced by dysregulation of the mammalian target of the rapamycin pathway (mTOR). This study aimed to examine the effects of continuous low-dose everolimus, a potent and selective inhibitor of mTOR, on the treatment of TSC-associated renal angiomyolipoma (AML). Between July 2013 and August 2017, 11 patients with TSC-AML were enrolled for an everolimus therapy protocol. An oral everolimus dose starting at 2.5 mg daily was gradually increased to 5.0 mg daily. All patients were evaluated using MRI or CT scanning at baseline, 12, 24, 36 and 48 months after the start of treatment for measuring changes of renal AML mass volume. Everolimus therapy resulted in significant shrinkage of TSC-AML volume after 48 months follow-up. Serum levels of everolimus were subdivided into group I (<8 ng/mL, n=6) and group II (>8 ng/mL, n=5). The volume reduction rates were 10.6%-65.2% in group I and 42.5%-70.6% in group II. To evaluate the response to treatment, three of six (50%) were responders in group I, and all the patients in group II (5/5, 100%) were responders. The differences in AML volume reduction between the groups were statistically significant at 12 months (p=0.011), 24 months (p=0006), 36 months (p=0.014) and 48 months (p=0.05). These results suggest that continuous low-dose everolimus therapy (2.5-5 mg daily) might be effective in shrinking TSC-AML volume and minimizes adverse effects and subsequent reducing medical costs.

Trend of seizure remission in patients with tuberous sclerosis complex: A retrospective medical review.

BACKGROUND: Seizures in tuberous sclerosis complex (TSC) tend to be intractable over time and become a subsequent psychological burden for the patients. The purpose of the current study was to describe the onset, phenotype, and factors associated with seizure remission in patients with TSC. METHODS: Patients diagnosed with TSC between 2009 and 2015 completed a questionnaire interview and underwent a systematic evaluation, including a medical review of their epilepsy history and neurobehavioral disorder assessment. RESULTS: Of the 61 patients, 50 patients (82.0%) had a positive seizure history. The active (n = 34) and seizure remission (n = 16) groups showed significant differences in age, neurobehavioral disorder, history of refractory epilepsy, and onset age (p < 0.001, p < 0.05, p < 0.05, and p < 0.05, respectively). The remission rates were 33.3% and 38.5% for those aged 6-18 years and over 18 years, respectively (p for trend = 0.01). CONCLUSION: Seizure remission can occur in adulthood. It shows a high correlation with patient age, minor refractory epilepsy, and neurobehavioral disorders.

Ensifer shofinae sp. nov., a novel rhizobial species isolated from root nodules of soybean (Glycine max).

Two bacterial strains isolated from root nodules of soybean were characterized phylogenetically as members of a distinct group in the genus Ensifer based on 16S rRNA gene comparisons. They were also verified as a separated group by the concatenated sequence analyses of recA, atpD and glnII (with similarities ≤93.9% to the type strains for defined species), and by the average nucleotide identities (ANI) between the whole genome sequence of the representative strain CCBAU 251167T and those of the closely related strains in Ensifer glycinis and Ensifer fredii (90.5% and 90.3%, respectively). Phylogeny of symbiotic genes (nodC and nifH) grouped these two strains together with some soybean-nodulating strains of E. fredii, E. glycinis and Ensifer sojae. Nodulation tests indicated that the representative strain CCBAU 251167T could form root nodules with capability of nitrogen fixing on its host plant and Glycine soja, Cajanus cajan, Vigna unguiculata, Phaseolus vulgaris and Astragalus membranaceus, and it formed ineffective nodules on Leucaena leucocephala. Strain CCBAU 251167T contained fatty acids 18:1 ω9c, 18:0 iso and 20:0, differing from other related strains. Utilization of l-threonine and d-serine as carbon source, growth at pH 6.0 and intolerance of 1% (w/v) NaCl distinguished strain CCBAU 251167T from other type strains of the related species. The genome size of CCBAU 251167T was 6.2Mbp, comprising 7,581 predicted genes with DNA G+C content of 59.9mol% and 970 unique genes. Therefore, a novel species, Ensifer shofinae sp. nov., is proposed, with CCBAU 251167T (=ACCC 19939T=LMG 29645T) as type strain.

The ultrastructural characterization of mitochondria-rich cells as a response to variations in salinity in two types of teleostean pseudobranch: milkfish (Chanos chanos) and Mozambique tilapia (Oreochromis mossambicus).

The pseudobranchs of two euryhaline teleost species, the milkfish (Chanos chanos) and the Mozambique tilapia (Oreochromis mossambicus), were studied after acclimization to different salinities using optical and electron microscopy. The milkfish pseudobranch was the lamellae-free type, with separate lamellae along the filaments containing two groups of mitochondria (Mt)-rich cells: chloride cells (CCs) and pseudobranch type cells (PSCs). Conversely, the tilapia pseudobranch was the embedded type, covered with connective tissues and with only one group of Mt-rich PSCs. Chloride cells were identified according to the apical openings and branched tubular networks around randomly distributed and diversely shaped Mt. Pseudobranchs type cells, however, were characterized according to the orderly arrangement of parallel tubules around closely packed Mt; both the tubules and the Mt were distributed in the vascular side of the cell, but were absent from the apical region. Compared with those of seawater (SW)-acclimated milkfish, the pseudobranchial lamellae of freshwater (FW) specimens were longer on average, and the Mt of the CCs had fewer cristae, were less electron-dense, and were often vacuolated. The Mt in the PSCs of FW-acclimated milkfish and tilapia were larger and more electron-dense than those of their SW-acclimated counterparts; in addition, more tubules were found to aggregately surround the Mt and basolateral membranes in the PSCs of fish from the hypo-osmotic environment. Conversely, the PSCs of tilapia were periodic acid-Schiff (PAS)-positive, and Mt in PSCs were concentrated with more parallel arrays of the tubule system than those of milkfish. Therefore, salinity-dependent changes in the ultrastructures of PSCs suggest their potential role in energy metabolism of both lamellae-free and embedded pseudobranchs, whereas the PAS-positive staining characteristics suggest a role in releasing or storaging polysaccharides in the embedded pseudobranch. J. Morphol. 278:390-402, 2017. © 2017 Wiley Periodicals, Inc.

Effects of everolimus on tuberous sclerosis complex-associated renal angiomyolipoma: A preliminary report.

AIM: Tuberous sclerosis complex (TSC) presents with multisystem benign neoplasm induced by dysregulation of the mammalian target of rapamycin pathway. This study aimed to examine the effects of oral everolimus at either 2.5 or 5.0 mg daily on the treatment of TSC-associated renal angiomyolipoma (AML). METHODS: Between July 2012 and August 2015, patients with TSC-associated renal AML were selected for everolimus therapy protocol. An oral everolimus starting dose at 2.5 mg was administered daily, and was gradually increased to 5.0 mg daily. All patients were evaluated using magnetic resonance imaging or computed tomography scanning at baseline, 12, 24, and 36 months after the start of treatment for measuring the changes of renal AML mass volume. RESULTS: Eight patients were finally enrolled for analysis in this study. Everolimus treatment had a statistically significant effect on the renal AML volume reduction during follow-up (P < 0.05). Renal AML mass volume reduction rates were 10.5-45.3% in four patients with everolimus 2.5 mg and 40.7-73.1% in four patients with everolimus 5.0 mg daily; the difference was statistically significant between the two groups (P < 0.05). Longitudinal follow-up for response to everolimus showed volume reduction rates to be around 10.5-73.1% in the initial 6-24 months after everolimus treatment, which remained stable during follow-up up to 36 months. CONCLUSION: The results suggest that an oral everolimus is effective and provides a non-invasive way to treat TSC-associated renal AML, and patients are likely to require maintenance therapy to continue to derive benefit.

Does Maternal Vitamin D Deficiency Increase the Risk of Preterm Birth: A Meta-Analysis of Observational Studies.

There are disagreements among researchers about the association between vitamin D deficiency during pregnancy and preterm birth (PTB). Therefore, we conducted a meta-analysis of observational studies to evaluate this association. We performed a systematic literature search of PubMed, MEDLINE and the Cochrane Library through August 2015 with the following keywords: "vitamin D" or "cholecalciferol" or "25-hydroxyvitamin D" or "25(OH)D" in combination with "premature birth" or "preterm birth" or "PTB" or "preterm delivery" or "PTD" or "prematurity". Our meta-analysis of 10 studies included 10,098 participants and found that pregnant women with vitamin D deficiency (maternal serum 25 (OH) D levels < 20 ng/mL) experienced a significantly increased risk of PTB (odds ratio (OR) = 1.29, 95% confidence intervals(CI): 1.16, 1.45) with low heterogeneity (I² = 25%, p = 0.21). Sensitivity analysis showed that exclusion of any single study did not materially alter the overall combined effect. In the subgroup analyses, we found that heterogeneity was obvious in prospective cohort studies (I² = 60%, p = 0.06). In conclusion, pregnant women with vitamin D deficiency during pregnancy have an increasing risk of PTB.

Association between the growth rate of subependymal giant cell astrocytoma and age in patients with tuberous sclerosis complex.

PURPOSE: The most common neurological complications associated with tuberous sclerosis complex (TSC) include intractable seizures that begin in infancy and subependymal giant cell astrocytoma (SEGA) complicated by hydrocephalus with increasing age. Information on SEGA growth of TSC patients is limited. This study aimed to examine the TSC-SEGA growth rates by periodic neuroimaging. METHODS: This study evaluated the TSC-SEGA growth rates by serial neuroimaging. Fifty-eight patients with TSC underwent systematic evaluation, including a review of medical history and serial brain neuroimaging. RESULTS: While magnetic resonance imaging was more sensitive in detecting cortical tubers than computed tomography (73.1 vs. 0 %, p < 0.001), its efficacy in identifying intracranial lesions was comparable to that of computed tomography (96.2 vs. 100 %, p = 0.658). Significant tumor growth was observed in children (p = 0.012) and adults (p = 0.028) during follow-up periods, respectively (median for children 23.5 months, interquartile range 18-40 months and median for adults 23 months, interquartile range 12-34 months). Further, the SEGA growth rate in children was significantly higher than that in adults (75.6 vs. 16.5 %, p = 0.03). CONCLUSIONS: The results of the study show that SEGA has a significantly higher growth rate in children using serial follow-up brain imaging, suggesting the importance of performing follow-up neuroimaging at yearly intervals in childhood to identify and prevent potential comorbidities.

Comparisons of two types of teleostean pseudobranchs, silver moony (Monodactylus argenteus) and tilapia (Oreochromis mossambicus), with salinity-dependent morphology and ion transporter expression.

There are essentially four different morphological types of pseudobranchs in teleosts, including lamellae-free, lamellae semi-free, covered, and embedded types. In the euryhaline silver moony (Monodactylus argenteus), the pseudobranch belongs to the lamellae semi-free type, which is characterized by one row of filaments on the opercular membrane and fusion on the buccal edge. The pseudobranchial epithelium of the moony contains two types of Na(+), K(+)-ATPase (NKA)-rich cells: chloride cells (CCs) and pseudobranch-type cells (PSCs). Our results revealed increased expression of NKA, the Na(+), K(+), 2Cl(-) cotransporter (NKCC), and the cystic fibrosis transmembrane conductance regulator (CFTR) for Cl(-) secretion and CCs profiles in the pseudobranchs of seawater (SW)-acclimated silver moonies, which indicates the potential role of pseudobranchs containing CCs in hypo-osmoregulation. In contrast, the pseudobranch of the Mozambique tilapia (Oreochromis mossambicus) belongs to the embedded type, which is covered by the connective tissues and only contains PSCs but not CCs. No sign of NKCC and CFTR-immunoreactivity (IR) was found in the pseudobranchs of SW and freshwater (FW) tilapia. However, higher NKA protein expression and larger sizes of NKA-IR PSCs were found in the pseudobranchs of FW-acclimated tilapia. Moreover, in the FW-acclimated moony, NKA-IR PSCs also exhibited higher numbers and larger sizes than in the SW individuals. Taken together, similar responses in low-salinity environments in different types of pseudobranchs indicated that the salinity-dependent morphologies of PSCs might be involved in critical functions for FW teleosts.

Targeting of surface alpha-enolase inhibits the invasiveness of pancreatic cancer cells.

Pancreatic Ductal Adenocarcinoma (PDAC) is a highly aggressive malignancy characterized by rapid progression, invasiveness and resistance to treatment. We have previously demonstrated that most PDAC patients have circulating antibodies against the glycolytic enzyme alpha-enolase (ENO1), which correlates with a better response to therapy and survival. ENO1 is a metabolic enzyme, also expressed on the cell surface where it acts as a plasminogen receptor. ENO1 play a crucial role in cell invasion and metastasis by promoting plasminogen activation into plasmin, a serine-protease involved in extracellular matrix degradation. The aim of this study was to investigate the role of ENO1 in PDAC cell invasion. We observed that ENO1 was expressed on the cell surface of most PDAC cell lines. Mouse anti-human ENO1 monoclonal antibodies inhibited plasminogen-dependent invasion of human PDAC cells, and their metastatic spreading in immunosuppressed mice was inhibited. Notably, a single administration of Adeno-Associated Virus (AAV)-expressing cDNA coding for 72/1 anti-ENO1 mAb reduced the number of lung metastases in immunosuppressed mice injected with PDAC cells. Overall, these data indicate that ENO1 is involved in PDAC cell invasion, and that administration of an anti-ENO1 mAb can be exploited as a novel therapeutic option to increase the survival of metastatic PDAC patients.

Five new phorbol esters with cytotoxic and selective anti-inflammatory activities from Croton tiglium.

Five new phorbol esters, (four phorbol diesters, 1-4, and one 4-deoxy-4α-phorbol diester, 5), as well as four known phorbol esters analogues (6-9) were isolated and identified from the branches and leaves of Croton tiglium. Their structures were elucidated mainly by extensive NMR spectroscopic, and mass spectrometric analysis. Among them, compound (1) was the first example of a naturally occurring phorbol ester with the 20-aldehyde group. Compounds 2-5, and 7-9 showed potent cytotoxicity against the K562, A549, DU145, H1975, MCF-7, U937, SGC-7901, HL60, Hela, and MOLT-4 cell lines, with IC50 values ranging from 1.0 to 43 µM, while none of the compounds exhibited cytotoxic effects on normal human cell lines 293T and LX-2, respectively. In addition, compound 3 exhibited moderate COX-1 and COX-2 inhibition, with IC50 values of 0.14 and 8.5 µM, respectively.

Comparison of the detection rates of RT-PCR and virus culture using a combination of specimens from multiple sites for enterovirus-associated encephalomyelitis during enterovirus 71 epidemic.

Enterovirus (EV) 71 may cause severe neurological illness in the pediatric population. The present study aimed to compare the detection rates of reverse transcription-polymerase chain reaction (RT-PCR) with pan-EV/EV71 type- specific primers and virus culture (VC) for the identification of EV and EV71 using specimens from multiple sites. In total, specimens from throat/rectal swabs, cerebrospinal fluid (CSF), and blood from 66 patients diagnosed with EV encephalomyelitis were subjected to both RT-PCR and VC for detecting the presence of pan-EV and EV71. The results revealed that the positive RT-PCR rate was higher in throat swabs (60.6%) and rectum swabs (50.0%) than in CSF (16.7%) and blood (15.6%). The same trend was also observed in case of VC: throat swabs (22.7%), rectum swabs (10.6%), and blood (3.0%). The detection rate of EV encephalomyelitis by RT-PCR was 90.9% for all samples, 63.6% of which were subtyped as EV71. The detection rates of RT-PCR were superior to those of VC, and identification using specimens from throat/rectal swabs yielded higher positive results. These findings may help physicians to identify the etiologies at an early stage during EV71 epidemics and to make emergent medical decisions for minimizing patient morbidity and mortality.

Characterization and biological evaluation of six new dimeric lignans with an unusual α,ß-unsaturated ketone motif from Zanthoxylum simulans.

Investigation of the bark of Zanthoxylum simulans afforded six new dimeric lignans zanthpodocarpins C-H (1-6) bearing an unusual α,ß-unsaturated ketone group. The new structures of 1-6 were determined by using detailed spectroscopic analysis. All of the isolated compounds were examined for their inhibitory effects against rat joint synovial cell and splenocyte proliferation. Compounds 1-6 showed potent anti-inflammatory activities with IC50 values ranging from 18.6 to 36.1µM, and 13.8 to 74.3µM.

The lamellae-free-type pseudobranch of the euryhaline milkfish (Chanos chanos) is a Na(+), K(+)-ATPase-abundant organ involved in hypoosmoregulation.

In teleosts, the pseudobranch is hemibranchial, with a gill-like structure located near the first gill. We hypothesized that the pseudobranch of the milkfish might exhibit osmoregulatory ability similar to that of the gills. In this study, the obtained Na(+), K(+)-ATPase (NKA) activity and protein abundance profiles showed that these parameters were higher in the pseudobranchs of the seawater (SW)- than the freshwater (FW)-acclimated milkfish, opposite the situation in the gills. The pseudobranch of the milkfish contained two types of NKA-immunoreactive cells, chloride cells (CCs) and pseudobranch-type cells (PSCs). To further clarify the roles of CCs and PSCs in the pseudobranch, we investigated the distributions of two ion transporters: the Na(+), K(+), 2Cl(-) cotransporter (NKCC) and the cystic fibrosis transmembrane conductance regulator (CFTR). NKCC on the basolateral membrane and CFTR on the apical membrane were found only in pseudobranchial CCs of SW-acclimated individuals. Taken together, the results distinguished NKA-IR CCs and PSCs in the pseudobranch of milkfish using antibodies against NKCC and CFTR as markers. In addition, increases in the numbers and sizes of CCs as well as in NKA expression observed upon salinity challenge indicated the potential roles of pseudobranchs in hypo-osmoregulation in this euryhaline teleost.

Progression of periodontal inflammation in adolescents is associated with increased number of Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Fusobacterium nucleatum.

OBJECTIVE: The study aims to evaluate the change of related subgingival periodontopathogens among different stage of gingivitis in adolescent and assess the relationship between periodontopathogens and the progression of periodontal inflammation. METHODS: A total of 77 subgingival plaque samples from 35 adolescent individuals were divided into three groups including gingivitis group (mild, 15 samples; moderate, 16 samples; severe, 15 samples), chronic periodontitis group (15 samples) and healthy group (15 samples). Real-time PCR was used to quantitate Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Fusobacterium nucleatum in subgingival plaque samples. RESULTS: All species, except for F. nucleatum, were detected in samples from gingivitis and periodontitis groups in significantly greater number than in those from healthy group (P < 0.05). In gingivitis groups, the number of P. gingivalis, T. forsythensis, and F. nucleatum in moderate and severe gingivitis groups was significantly higher than in mild gingivitis group (P < 0.05). After merging moderate gingivitis and severe gingivitis groups into moderate-to-severe gingivitis group, the four periodontopathogens were detected in samples from periodontitis group in significantly greater number than in those from moderate-to-severe gingivitis group (P < 0.05). CONCLUSION: The number of P. gingivalis, P. intermedia, T. forsythensis, and F. nucleatum in subgingival plaque increases with progression of periodontal inflammation in adolescents. Examination of periodontopathogens number in adolescents may be of some value for monitoring of periodontal disease development.

[Symbiotic matching between soybean cultivar Luhuang No. 1 and different rhizobia].

Soybean plants could establish symbiosis and fix nitrogen with different rhizobial species in the genera of Sinorhizobium and Bradyrhizobium. Studies on the symbiotic matching between soybean cultivars and different rhizobial species are theoretically and practically important for selecting effective strains used to inoculate the plants and improve the soybean production and quality. A total of 27 strains were isolated and purified from a soil sample of Huanghuaihai area by using the soybean cultivar Luhang No. 1, a protein-rich cultivar grown in that area, as the trapping plants. These strains were identified as members of Sinorhizobium (18 strains) and Bradyrhizobium (9 strains) based on the sequence analysis of housekeeping gene recA. Two representative strains (Sinorhizobium fredii S6 and Bradyrhizobium sp. S10) were used to inoculate the seeds of Luhang No. 1 alone or mixed, in pots filled with vermiculite or soil, and in the field trial to investigate their effects on soybean growth, nodulation, nitrogen fixation activity, yield, contents of protein and oil in seeds. The results demonstrated that strain S6 showed better effects on growth-promotion, yield of seeds and seed quality than strain S10. Thus strain S6 was finally regarded as the effective rhizobium matching to soybean Luhuang No. 1, which could be the candidate as a good inoculant for planting the soybean Luhuang No. 1 at a large scale in the Huanghuaihai area.

[Study on chemical constituents of Zanthoxyli cortex's ethyl acetate extract].

OBJECTIVE: To study the chemical constituents of Zanthoxyli Cortex. METHODS: The chemical constituents were isolated and purified by silica gel and HP-20, MCI gel, Sephadex LH -20 column chromatography, RP-18 and PTLC. Their structures were elucidated by the analysis of spectral data and chemical properties. RESULTS: Ten compounds were isolated from EtOAc extract and their structures were identified as: asarinin (I), fargesin (II), eudesmin (III), (1R, 2R, 5R, 6S)-2-(3,4-dimethoxyphenyl)-6-(3,4-dihydroxyphenyl)-3,7-dioxabicyclo[3.3.0]-octane(IV), dimethoxysamin(V), rel-(1R,5R,6S)-6-(3,4-dimethoxyphen-yl)-3,7-dioxabicyclo-[3.3.0]-octan-2-one(VI), Magnone A(VII), beta-sitoste-rol( VIII), beta-armyrin(IX), beta-amyrone(X). CONCLUSION: These compounds isolated from Zanthoxyli Cortex's Ethyl acetate extract are all known compounds. Fargesin(II) and beta-amyrone(X) are isolated from Zanthoxyli Cortex for the first time.

[Recombinant plasmid ZLW/pEGFP-C2 transfection into schistosomula of Schistosoma japonicum].

OBJECTIVE: To study the efficiency of ZLW/pEGFP-C2 plasmid transfection into Schistosoma japonicum schistosomula and observe its in vitro effect of anti-schistosomula. METHODS: Recombinant plasmid ZLW/pEGFP-C2 was transfected into mechanically transformed schistosomula by immersion in 0.75% DMSO and high concentration of plasmid. Enhanced green fluorescent protein (EGFP) transfected cells were observed under inverted fluorescence microscope. At 48 hours after culture, total RNA and proteins from transfected schistosomula were extracted, and the presence of the transgenes (ZLW and EGFP) in schistosomula were confirmed by RT-PCR and Western blotting. At 24, 48, 72, and 96 hours after transfection, the schistosomula were counted by light microscope with methylene blue staining. pEGFP-C2 empty plasmid group and TBS group served as controls. RESULTS: The transfection rate was about 10%. The fluorescence of ZLW/EGFP protein was mainly localized in the tegument of the worms, especially abundant around oral sucker and ventral sucker. The expected size of 259 bp fragment was successfully amplified by RT-PCR and confirmed by DNA sequencing. Western blotting analysis showed that ZLW/EGFP was expressed in schistosomula. No statistically significant difference was established for schistosomula mortality among ZLW/pEGFP-C2 group (14.0%, 48.8%), pEGFP-C2 group (15.9%, 45.7%) and TBS group (16.9%, 50.3%) at 24 and 48 hours after transfection (P > 0.05). At 72 hours after transfection the mortality rate of ZLW/pEGFP-C2 group (92.7%) was significantly higher than that of pEGFP-C2 group (73.2%) (P < 0.01), and after 96 h the mortality in ZLW/pEGFP-C2 group increased to 100%. CONCLUSION: ZLW/pEGFP-C2 plasmid has been introduced into juvenile S. japonicum by immersion in 0.75% DMSO and high concentration of plasmid, and was expressed in the parasite.