Objective To prokaryotically express the valosin-containing protein(VCP)of Schistosoma japonicum,and ana-lyze its VCP mRNA expressions in the cercaria,schistosomulum,adult worm(female and male worms)and egg. Methods RNA of S. japonicum eggs were extracted,and reversely transcribed into cDNA. The VCP gene of S. japonicum was amplified by using polymerase chain reaction(PCR),and subcloned into the prokaryotically expressed vector pET15b. The recombined plasmid was transformed into BL21 cells,and the expression of the target gene was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG). The recombinant protein was yielded through the purification of inclusion body,and identified by using sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The RNA(s)of cercaria,schistosomulum,female adult worm,male adult worm,and egg of S. japonicum were extracted,digested with DNase,purified,and reversely transcribed into cDNA. The mRNA expressions of the VCP gene in various developmental stages of S. japonicum were determined by using fluorescence-based quantitative real-time PCR. Results The VCP gene of S. japonicum was yielded by PCR amplification,and the recombinant pro-tein was obtained through recombinant plasmid expression and purification of inclusion body. The highest VCP mRNA expression in S. japonicum cercaria was detected by the fluorescence-based quantitative real-time PCR,while low expressions were found in the schistosomulum,egg,female and male adult worms. Conclusion The recombinant protein encoded by the VCP gene of S. ja-ponicum is successfully obtained,and the VCP mRNA expression is determined in various developmental stages of S. japonicum.

Objective To enhance the protective immunity effects against Schistosoma japonicum infection by priming with cocktail DNA vaccines and boosting with cocktail protein vaccines in infected BALB/c mice.Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml,and mixed with pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs by equal volume to form the cocktail DNA vaccine,and also mixed with recombinant proteins SjC23-HD,SjCTPI,and NP30 by equal volume to form the cocktail protein vaccine.Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups(A,B,C,D,E).In Group A(control group),each mouse was immunized with 100 ?l saline solution by intramuscular(i.m.);in Group B(pcDNA3.1 control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6;in Group C(pcDNA3.1 and cocktail protein group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccines group),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6;in Group E(cocktail DNA vaccines plus cocktail proteins),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9.Four weeks after the last DNA immunization or two weeks after protein boosting,all the mice were challenged with(40?1)cercariae of Schistosoma japonicum by abdominal skin penetration at the same time.Forty-two days post-challenge,the mice were sacrificed and perfused,and the numbers of recovered worms and eggs in liver were counted.The blood was collected from the tail veins of all the mice two days before the first immunization and challenge,respectively,the serum was prepared for detection of IgG,IgG1 and IgG2a.Two days before the challenge,the spleen cells of two mice from each group were cultured and stimulated with ConA and soluble egg antigen(SEA),and the supernatant was collected for detection of IL-2,IL-4 and IFN-?.Results The worm reduction rates in Group C,D and E were 17.70%,32.88% and 45.35%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P

Objective To know the contamination status of Giardia lamblia and Cryptosporidium in drinking water of Jiangsu Province,so as to provide the evidence for producing hygiene and safety drinking water. Methods A total of 28 water plants of 13 cities in Jiangsu Province were selected,and the source water(10 L),chlorinated water(100 L)and tap water(100 L) were collected separately in each site. The water samples were then treated by filtration,washing,centrifuging concentration, immune magnetic separation,and immunofluorescent assay,to detect the numbers of Giardia cysts and Cryptosporidium oocysts. Results Totally 84 samples from 13 cities were collected,including 28 source water,28 chlorinated water and 28 tap water samples. Among the chlorinated water and tap water samples,no Giardia cysts and Cryptosporidium oocysts were found. Howev-er,Giardia cysts were detected in 3(10.71％,3/28)source water samples(Yancheng,Lianyungang,Changzhou cities),with the density of 1 cyst/10 L of all. Cryptosporidium oocysts were also detected in 3(10.71％,3/28)source water samples(Nan-jing,Zhenjiang,Yangzhou cities),with the density of 1 oocyst/10 L of all. Conclusions The source water in partial areas of Ji-angsu Province has been contaminated by Giardia and Cryptosporidium. To ensure the safety of drinking,the regulation of source water and surveillance of drinking water should be strengthened.

Objective To study the protective effect of codon optimized TPI DNA vaccine against Schistosoma japonicum infection.Methods Sixty female BALB/c mice were randomly divided into 5 groups.The mice were injected through musculus quadriceps fexoris with 100 ?g pcDNA 3.1 control(Group A), pcDNA3.1-TPI(Group B), pcDNA 3.1-TPI-mHSP70(Group C), pcDNA3.1-TPI.opt(Group D), and pcDNA3.1-TPI.opt-mHSP70(Group E) respectively.All mice were immunized for three times with an interval of two weeks.The mice were challenged with(40?1) cercariae of S.japonicum per mouse by abdominal skin penetration 4 weeks after the last immunization, and sacrificed at 42 days post-challenge, the number of worms or hepatic eggs was counted.Blood was taken for the detection of IgG, IgG1, and IgG2a 2 days before immunization and before challenge, respectively.Spleen cells of 2 mice from each group were cultured and stimulated with ConA and rSjCTPI peptide, and the supernatant was collected for detection of IL-2, IL-4, IL-5, IFN-?, and TNF by flow cytometry.Results ELISA showed that the mice in groups B, C, D, and E produced specific IgG and IgG1, IgG2a antibody isotypes, and the ratio of IgG2a/IgG1 was 1.73, 2.06, 2.44, and 3.09, respectively.The levels of IL-2, IFN-? and TNF in groups D and E were higher than that of groups B and C.The worm reduction rate and hepatic egg reduction rate in groups D(36.03%, 41.7%) and E(39.03%, 46.85%) were higher than those of groups B(26.28%, 28.35%) and C(28.38%, 31.39%)(P

Objective To investigate the correlation between reproductive hormone concentration and the amplitude and latency of speech-evoked auditory brainstem response (speech-ABR) in young adults, and to explore the effects of reproductive hormone on the speech processing ability of young people.Methods Speech-ABR of thirty five normal hearing young adults, including seventeen females (27.29±1.83 years old) and eighteen males (28.17±2.50 years old) were recorded.The speech syllable /da/ was transmitted as a stimulus sound to the right ears through insert earphones in speech-ABR test.All participants had air conduction hearing thresholds of 20 dB HL or better across the standard audiometric frequencies (250~8 000 Hz) in both ears, and click-ABRs were also within normal limits.At the same time, the concentrations of estradiol and testosterone in the serum were examined.Results ① Females had a shorter latency than males in transient responses (waves V, A and O) and sustained responses (waves D, E and F) of speech-ABR (P<0.05, respectively).The amplitude of transient response (waves V and A) and sustained response (waves D, E and F) in females was also significantly larger than that in males (P<0.05, respectively), except for amplitude of peak O (P>0.05).The V/A slope in females was significantly steeper than that in males (P<0.05).② Estradiol levels in females (118.77±102.66 pg/ml) were significantly higher than that in males (52.91±14.77 pg/ml) (P<0.05), and the total testosterone concentration in females (457.65±140.82 pg/ml) was significantly lower than that in males (3 677.37±1 155.80 pg/ml) (P<0.05).③ A correlation analysis between speech-ABR and estradiol or total testosterone showed that all peak latencies of speech-ABR in transient responses (waves V, A and O) and sustained responses (waves D, E and F) were negatively correlated with the estradiol concentration (P<0.05 respectively), in which the correlation coefficient was between 0.2~0.4.All peak latencies of speech-ABR were positively correlated with the total testosterone concentration (P<0.05 respectively), in which the correlation coefficient was between 0.4~0.7.④ The amplitudes of speech-ABR increased with estradiol concentration growing, the wave V and estradiol concentrations were positively correlated (P<0.05).The estradiol concentrations showed a significant negative correlation with wave A, D, E, F and O wave (P<0.05 respectively), with a correlation coefficient between 0.2~0.7.On the contrary, the amplitudes of speech-ABR decreased with the increasing of total testosterone concentration, and the wave V, wave A, V/A slope and total testosterone concentration were moderately correlated (P<0.05),with a correlation coefficient between 0.4-0.6.The correlation between the amplitudes of D wave and total testosterone concentration was not statistically significant (P>0.05), and the correlation between wave E and wave F and total testosterone concentration was weakly correlated (P<0.05).In addition, the amplitudes of the wave O were also independent with testosterone levels (r=0.133, P>0.05).Conclusion There are correlations between the level of reproductive hormone and the amplitude and latency of speech-ABR.It is one of the reasons for the gender difference in the brainstem speech coding ability of normal young adult.

Objective To enhance the protective immunity effects of nucleic acid vaccines against Schistosoma japonicum infection by electroporation(EP)in vivo in infected BALB/c mice.Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml.pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs were mixed by equal volume to form the cocktail DNA vaccine,and also mixed with recombinant proteins SjC23-HD,SjCTPI,and NP30 by equal volume to form the cocktail protein vaccine.Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups(A,B,C,D,E).In Group A(control group),each mouse was immunized with 100 ?l saline solution by intramuscular(i.m.);in Group B(pcDNA3.1/EP control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 followed by EP in vivo for three times at week 0,3,6;in Group C(pcDNA3.1/EP plus cocktail protein vaccine group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 followed by EP for three times at week 0,3,6 and boosted with 100 ?l cocktail protein vaccine plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccine/EP group),each mouse was immunized(i.m.)with 100 ?l cocktail DNA vaccine followed by EP for three times at week 0,3,6;in Group E(cocktail DNA vaccine/EP plus cocktail protein vaccine group),each mouse was immunized(i.m.)with 100 ?l cocktail DNA vaccine followed by EP for three times at week 0,3,6 and boosted with 100 ?l cocktail protein vaccine plus 100 ?l FCA by subcutaneous at week 9.Four weeks after the last DNA immunization or two weeks after protein boosting,all the mice were challenged with(40?1)cercariae of Schistosoma japonicum by abdominal skin penetration.Forty-two days post-challenge,the mice were sacrificed and perfused,and the numbers of recovered worms and eggs in liver were counted.The blood was collected from the tail veins of all the mice two days before the first immunization and challenge,respectively,the serum was prepared for detection of IgG,IgG1 and IgG2a.Two days before the challenge,the spleen cells of two mice from each group were cultured and stimulated with ConA and soluble egg antigen(SEA),and the supernatant was collected for detection of IL-2,IL-4 and IFN-? by flow cytometre.Results The worm reduction rates in Group C,D and E were 18.09%,45.00% and 57.09%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P

OBJECTIVETo evaluate the value of the clinical use of CT perfusion imaging (CTPI) and CT subtraction angiography (CTSA) for diagnosing acute ischemic cerebrovascular disease (AICVD).

METHODSTwenty-four patients with AICVD onset within 24 hours were examined with regular CT, CTPI, and CTSA. Some cases received CTPI, magnetic resonance imaging (MRI), magnetic resonance angiography (MRA), digital subtraction angiography (DSA) or single photon emission computer tomography (SPECT) during follow-up examinations.

RESULTSOf the 24 cases, 11 had negative results from regular CT scans 3 - 6 hours after onset of stroke in 6 cases, 6 - 12 hours in 3 cases, and 12 - 24 hours in 2 cases. Ten of these cases were then confirmed by CTPI as having ischemic lesions, 2 with middle cerebral artery occlusion (MCAO), and 1 case with transient ischemic attack (TIA) with CTPI negative. Of the 24 cases, 13 had positive results from regular CT, 9 were diagnosed with ischemic lesions larger by using CTPI than regular CT, 1 case had MCAO and 1 had internal carotid artery occlusion (ICAO). There were 4 cases with ischemic lesions observed with regular CT having nearly the same range as that of lacunar infarctions using CTPI. Another 4 cases had more than 2 lesion areas. The peak time (PT), mean transit time (MTT) and relative flow (RF) of 24 cases were markedly different. The sides of ischemic lesions compared to each other and the core of the lesion compared to peripheral zones were also altered significantly (P < 0.01).

CONCLUSIONSCombined CTPI with CTSA can detect acute ischemic lesions at early and hyper-early stages and could distinguish between TIA, lacunar infarction and a larger area of infarction. Using semiquantitative blood perfusion analysis status, CTPI with CTSA could define position, area and range of the ischemic lesion and penumbra. These scans can also analyze the brain blood perfusion status. It is important to early diagnose the occlusion of the entire division of the internal carotid artery or middle cerebral artery and it is meaningful to assess prognosis and assignment of therapy.

Adult ; Aged ; Angiography, Digital Subtraction ; Brain Ischemia ; diagnostic imaging ; Cerebral Angiography ; Cerebrovascular Circulation ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed

Objective: This study was designed to identify the relative factors, bacteriological profile and their antibiotic susceptibility pattern of neonatal sepsis. Methods: A retrospective survey was conducted on the clinical information, pathogen identification and antibiotic sensitivity results of 425 newborns with neonatal sepsis admitted to Nanjing Maternity and Child Health Care Hospital from 2010 to 2017. Of the 425 positive blood-cultures, 148 (34.82%) were early-onset neonatal sepsis (EOS) and 277 (65.18%) were late-onset neonatal sepsis (LOS). Clinical information and pathogen identification were compared between EOS and LOS. Antibiotic sensitivity of gram negative organisms (G^-) and gram positive organisms (G⁺) were also detected. Results: The rates of premature delivery (78.70%, n=218), low birth weights (67.15%, n=186) and cesarean delivery (59.57%, n=165) were significantly increased in LOS (P<0.05) compared with those rates in EOS, which were 41.89% (n=62), 37.84% (n=56) and 46.62% (n=69). Parturients fever (18.24%, n=27) and meconium like amniotic fluid (25.68%, n=38) were significantly increased in EOS (P<0.05) compared with those rates in LOS, which were 7.94% (n=22) and 5.42% (n=15). Among the identified pathogen, the incidence of G^- and G⁺ bacteria were 216 (50.83%) and 201 (47.29%) respectively, and the rest was Candida glabrata (1.88%, n=8). Escherichia coli 68 (16.00%) was the predominant isolate followed by Klebsiella pneumoniae (13.18%, n=56). The detection rate of Hemolytic staphylococcus (10.81%, n=16) was significantly increased in EOS (P<0.001) compared with LOS (1.44%, n=4). However, the incidence of Klebsiella pneumonia (5.88%, n=44) was higher in LOS (P=0.024) compared with EOS (8.11%, n=12). Most of the gram positive isolates exhibited high resistance to penicillin (90.32%-100.00%) and cephalosporin group antibiotics (25.00%-100.00%). Similarly, the majority of the gram negative isolates showed higher resistance to ampicillin (83.33%-100.00%), but susceptible to aminoglycosides (0-11.76%), quinolones (0-17.65%) and β-lactams (0-5.88%). Conclusion Among the study population, the percent of preterm, low birth weight and cesarean section were higher in LOS while parturients fever and meconium-like amniotic fluid were higher in EOS. The pathogens with the highest detection rate were Escherichia coli and Klebsiella pneumoniae. The results of antibiotic susceptibility test showed that common pathogens had high resistance to commonly used antibacterial drugs.

Objective To investigate the core emergency response capabilities of medical workers for corona virus disease 2019 (COVID-19) public health emergency, in order to provide references for improving emergency level. Methods By means of convenient sampling, the general information questionnaire and the medical staff's 2019-new coronavirus (2019-nCoV) core emergency capability questionnaire were used to conduct this survey, and the status of 2019-nCoV core emergency capability of 629 clinical medical staff was analysed. Results The score of core emergency response capability of medical staff for 2019-nCoV was (135.60±21.73)points, with a score rate of 73.3%; the score of preventive capability dimension was (11.69±1.96)points, with a score rate of 77.93%; the score of preparation ability dimension was (20.79±4.18)points, and the score rate was 69.30%; the score of rescue ability was (103.11±16.93)points, with a scoring rate of 73.57%. Conclusion The core emergency response capability of medical worker for 2019-nCoV and its dimensions are at a medium level. At the present stage, the training of core knowledge of medical personnel 2019-nCoV should be strengthened, effective measures should be taken to improve the alleviation of weakness in rescue, and the self-psychological adjustment of medical personnel should be strengthened to improve the core emergency response capability of medical worker.

Objective To construct a vimentin glycosylated mutant plasmid,and to study its impact on rat adrenal chromaffin cell (PC12) cell differentiation.Methods The vimentin glycosylated mutant plasmid was constructed,and the gel electrophoretic was performed and the sequencing identification was performed. The empty plasmid pcDNA3.1b (control group),vimentin plasmid (Vim group) and vimentin glycosylated mutant plasmid (Vim mut group) were used to transfect PC12 cells,which were cultured for 0,3 d in the con-dictions of treatment and non-treatment of exogenous oligosaccharides (Cyclo-ManN pro) respectively,and the cell differentiation records and quantitative analysis (the cell average neurite length and differentiated neu-rite cells percentage) were performed.Results A recombinant plasmid containing a glycosylation site muta-tion site 19(T-G),97(A-G),100(T-G) was successfully constructed (about 1404 bp in size).Before and after Cyclo-ManN pro treatment,the average neurite length of PC12 cells and the percentage of differentiated neu-rite cells in the Vim group were greater than those in the control group and the Vim mut group[(61.98±19.03)μm vs.(51.09±14.45)μm,(51.49±14.78)μm,(6.60±0.25)％ vs. (4.27±0.18)％,(4.76±0.33)％;(78.01±18.31)μm vs. (69.98±12.85)μm,(68.45±13.84)μm,(10.62±0.25)％ vs. (8.11±1.22)％,(5.89±0.60)％],the difference was statistically significant (P<0.05).Conclusion The glycosyla-tion modification of vimentin could be used for PC12 cell differentiation.