Clinical effect of two fixation methods by anterolateral approach in the treatment of Rüedi-Allg?wer Ⅱ Pilon fracture / 中国医师进修杂志

Objective:To investigate the clinical effect of primary debridement combined with external fixation and secondary sequential internal fixation in the treatment of Rüedi-Allg?wer Ⅱ Pilon fracture.Methods:The clinical data of 36 patients with Rüedi-Allg?wer Ⅱ Pilon fracture from January 2017 to December 2019 in the People′s Hospital of Quzhou City, Zhejiang Province were retrospectively analyzed. Among them, 16 patients were treated with primary debridement with calcaneal traction and secondary internal fixation (internal fixation group), and 20 patients were treated with primary debridement combined with external fixation and secondary sequential internal fixation (combined fixation group). The operative time, intraoperative blood loss, postoperative drainage volume, time of full weight bearing, fracture healing time, American Society of Foot and Ankle Surgery (AOFAS) posterior ankle foot function score, visual analogue score (VAS), reduction quality (Burwell-Charnley score) and incidence of complication were compared between 2 groups.Results:The patients were followed up for 6 to 18 (10.7 ± 2.8) months. The time of full weight bearing and fracture healing time in combined fixation group were significantly shorter than those in internal fixation group: (7.2 ± 1.9) weeks vs. (9.4 ± 2.1) weeks and (3.4 ± 0.8) months vs. (4.1 ± 1.2) months, and there were statistical differences ( P<0.05 or <0.01); there were no statistical differences in operative time, intraoperative blood loss and postoperative drainage volume between 2 groups ( P>0.05). There was no statistical difference in AOFAS posterior ankle foot function score 1 month after surgery between 2 groups ( P>0.05); the AOFAS posterior ankle foot function score 3 and 6 month after surgery in combined fixation group was significantly higher than that in internal fixation group: (86.4 ± 1.7) scores vs. (75.7 ± 1.2) scores and (93.6 ± 2.2) scores vs. (82.1 ± 1.9) scores, and there was statistical difference ( P<0.05). There was no statistical difference in VAS between 2 groups ( P>0.05). There were no statistical differences in rate of reduction satisfaction and incidence of complication between 2 groups ( P>0.05). Conclusions:The primary debridement combined with external fixation and secondary sequential internal fixation for the treatment of Rüedi-Allg?wer Ⅱ Pilon fracture is conducive to the rapid recovery, which is worthy of extensive clinical promotion.

Immediate ear reconstruction by superficial temporal fascia flap combined with avulsion auricular tissue / 中华整形外科杂志

Objective@#To explore the application and effect of superficial temporal fascia flap combined with avulsion auricular tissue in emergency auricular restoration.@*Methods@#From June 2015 to December 2015, 6 patients with auricular large area complete avulsion were underwent treatment in Department of Plastic Surgery of General Hospital of Shenyang Military. After thorough debridement, the auricular cartilage scaffold of the avlusion ear and skin was completely stripped. The auricular cartilage was repositioned on its anatomical site and subsequently covered by superficial temporal fascia flap. The free skin was stripped as full-thickness graft to cover the surface of reconstructed ear.@*Results@#All 6 patients with auricle large area complete avulsion achieved immediate repair under emergency condition. The operations were successfully completed and the ears were healed primarily. The patients were followed-up for one year. Five patients with partial auricular avulsion achieved obvious reconstructed auricle profile. The color of reconstructed ear was close to the surrounding skin and the cranioauricular angle was nearly normal. Patients and their families were very satisfied. One patient of total auricular reconstruction had auricular contracture. The auricle profile was not obvious with small size, morphological changes and external auditory canal stenosis.@*Conclusions@#Avulsion auricle and temporal superficial fascia flap can be used to repair partial auricle defects as a first-stage repair with ideal results. It is the best choice for large auricle defects in emergency cases.

Cloning, eukaryotic expressing and function analysis of Schistosoma japonicum apoptosis gene Sjcaspase3 / 生物工程学报

For further research of the apoptosis mechanism of Schistosoma japonicum (S. japonicum). The cDNA encoding Sjcaspase3 of Schistosoma japonicum was amplified by polymerase chain reaction (PCR) technique, which contained 900 nucleotides and encoded 299 amino acids. The theory molecular weight and isoelectric point (PI) of the deduced protein is 33.5 kDa and 6.39, respectively. Real-time PCR was used to analyze the transcription profiles of Sjcaspase3 at different development stages of S. japonicum. The results showed that this gene was expressed in all stages of S. japonicum with the highest expression in 21d worms, and the level of gene transcription in 42 d female worms was higher than that of male worms. The recombinant plasmid pXJ40-FLAG-Sjcaspase3 was constructed and transfection into Hela cells successfully. Real-time PCR and Western blotting analysis showed Sjcaspase3 was successfully expressed in Hela cells. Enzyme activity analysis revealed that recombinant Sjcaspase3 possessed the activity to cut substrate DEVD. Flow cytometry proved that Sjcaspase3 could induce early apoptosis of Hela cells. The results provide the basis for proceeding further study on the biological function of Sjcaspase3 and better understand the apoptosis mechanism of S. japonicum.

Effects of early drainage tube occlusion on blood loss after total knee arthroplasty / 中国组织工程研究

BACKGROUND:Artificial knee joint replacement in older patients often combines with basic diseases, such as hypertension and diabetes. Perioperative blood loss is an important factor affecting the safety of replacement. OBJECTIVE: To explore the effect of the early closure of drainage tube on blood loss after primary total knee arthroplasty. METHODS: We randomly selected 90 patients with osteoarthritis of the knee who underwent primary total knee arthroplasty in the Affiliated Hospital of Qingdao University from January 2014 to July 2015. The patients were randomly divided into three groups (n=30). In the 4-hour occlusion group, the drainage tube was closed for 4 hours in early stage of replacement. In the 2-hour occlusion group, the drainage tube was closed for 2 hours in early stage of replacement. In the control group, the drainage tube was not closed. Because of the use of tourniquet during surgery, the amount of intraoperative blood loss was considered as 0 mL. Drainage blood loss after surgery was recorded. Total blood loss was calculated according to Gross formula through patient height, weight and preoperative and postoperative hematocrit. Hidden blood loss was gotten by subtracting the visible blood loss from total loss. Under the observation of postoperative joint sweling and subcutaneous ecchymosis, knee Hospital for Special Surgery score was recorded at 6 weeks after replacement, and compared among groups. RESULTS AND CONCLUSION:Statistical analysis indicated that significant differences in total blood loss and dominant blood loss were detected among the three groups (P 0.05). The incidence of joint sweling and subcutaneous ecchymosis was increased in the 4-hour occlusion group (P < 0.05). Above results confirmed that drainage tube occlusion can decrease total blood loss and dominant blood loss after total knee arthroplasty, but cannot reduce hidden blood loss. 2-hour occlusion after total knee arthroplasty is an ideal choice, but the amount of hidden blood loss should be carefuly considered.

Clinical effect of laparoscopy combined with choledochoscopy on choledocholithiasis / 中华老年医学杂志

Objective To explore the clinical effect of laparoscopy combined with choledochoscopy on choledocholithiasis.Methods Totally 134 elderly patients with choledocholithiasis were treated in our hospital from Jan 2013 to Dec 2014, who were randomly divided into observation group and control group (n=67 for each), treated with laparoscopy combined with choledochoscopy, and traditional surgery, respectively.The operation time, bleeding volume, exhaust time, in-hospital stay, complications and residual stones rate were compared between the two groups.Results The operation time was higher in observation group than in control group [(124.6±21.2) min vs.(94.7± 17.9) min, t=8.821, P＜0.001].The bleeding volume were less in observation group than in control group[(43.8±10.4) ml vs.(113.5±37.6) ml, t=14.624, P＜0.001].The exhaust time and in hospital time were decreased in observation group than in control group[(27.6 ±5.5) h vs.(43.4±8.1) h, (7.4±2.4) d vs.(10.3±2.8) d, t=13.209 and 6.437, P＜0.001 for both].The incidences of postoperative pain and other complications were lower in observation group than in control group [6.0％ vs.28.4％, 16.4％ vs.43.3％, x2=11.810and 11.547, P=0.001 for all].Conclusions The laparoscopy combined with choledochoscopy has advantages to minimize the surgical injury, reduce the bleeding volume and promote the postoperative recovery in treating choledocholithiasis in elderly patients.

Comparison of clinical data of lung cancer between elderly and middle-aged patients accepting operations / 中国癌症杂志

Background and purpose:The morbidity and mortality of lung cancer are currently the highest malignant tumor in China and the world. Most onset age of the illness is after 60 years old. Therefore, surgical resection of lung cancer in the elderly is very worthy of concern. This study aimed to investigate the clinical features, epidemic characteristics and conditions in perioperative period between the elderly and middle-aged patients with lung cancer, and provide the reference for clinical treatment.Methods:Totally 1 019 patients with lung cancer who were admitted to the hospital and accepted the operations in department of thoracic surgery in Shanghai Pulmonary Hospital from Jan. 2007 to Dec. 2012 were analysed retrospectively. The clinical data including gender, pathological type, TNM stage, intraoperative amount of bleeding and post-operative length of hospitalization were compared.Results:There was a signiifcant difference in sexual factors between these two groups (P?0.05), and compared with the middle-aged group, the proportion of male was more higher in the elderly group (76.91%vs 52.81%). Adenocarcinoma was the most common and squamous carcinoma was the next in both two groups. The constituent ratio of the pathlogical type between the elderly group and the middle-aged group was statistically signiifcant (P?0.05). The squamous carcinoma in the elderly group was higher than that in the middle-aged group (37.5%vs 15.6%). On the contrary, adenocarcinoma was more common in the middle-aged group (72.8%vs 50.7%). StagesⅡa,Ⅱb, andⅢa were more common in the elderly group and stagesⅠa, andⅠb were the most clinical stage in the middle-aged group. The clinical stage between two groups was statistically significant (P?0.05).The intraoperative amount of bleeding was higher and the post-operative length of hospitalization was longer in the elderly group, with a signiifcant difference as compared with that in the middle-aged group(P?0.05). And there was a signiifcant difference in incidence of accompanying diseases between the two groups, compared with the middle-aged group, the proportion of accompanying diseases was more higher in the elderly group(58.6%vs42.3%).Conclusion:Elderly patients with lung cancer are more common in males, with adenocarcinoma being the most common. The cancer mostly belongs to a medium or advanced stage. Elderly patients have the trend with more amount of bleeding in operation and lengh of stay.

Cloning, expression of gene SjOST48 from Schistosoma japonicum and evaluation of the immunoprotective efficacy of rSjOST48 in mice / 生物工程学报

To identify SJCHGC01743 gene of Schistosoma japonicum and evaluate the potential of the recombinant protein as a new vaccine candidate for schistosomiasis, polymerase chain reaction (PCR) technique was used to amplify the cDNA of the gene and real-time RT-PCR was used to analyze the transcription profiles of SJCHGC01743 at different development stages. Recombinant plasmid was successfully constructed and transformed into competent Escherichia coli BL21 (DE3). Then the recombinant protein was expressed, purified and emulsified with ISA206 adjuvant to immunize BALB/c mice for three times. The immunogenicity was confirmed by Western blotting and tissue localization was detected by indirect immunofluorescent assay. The specific antibody level was detected by ELISA. The immunoprotection of rSjOST48 was evaluated by the reduction in worm and egg counts in mice. A cDNA with 1 248 nucleotides was isolated from 28-day-old schistosomes cDNAs by PCR. Sequence analysis revealed that SJCHGC01743 was a 48-kDa subunit of the oligosaccharyltransferase complex (OST48) and named as SjOST48. Real-time PCR analysis indicated that this gene was expressed in all investigated stages and had the highest expression level in 28 d worms, the level of gene transcription in female worms was significantly higher than that of male worms. Then recombinant plasmid pET28a(+)-SjOST48 was successfully constructed and expressed in E. coli BL21 (DE3). Western blotting analysis showed that rSjOST48 had good immunogenicity. Indirect immunofluorescent analysis revealed that SjOST48 was mainly distributed on the tegument of the worms. The result of ELISA indicated that the rSjOST48 vaccinated group could induce a significant increase in the level of specific IgG, IgG1 and IgG2a. An immunoprotection experiment showed that the vaccination of rSjOST48 in mice induced 32.62% (P < 0.05) reduction in the numbers of worms and 57.61% (P < 0.01) in eggs in liver, compared with that of the control group. This study provides the foundation for proceeding further research on the biological function of SjOST48 and screening new vaccine candidates for schistosomiasis.

Cloning, expression and protective efficacy evaluation of radiation sensitive protein 23 (RAD23) from Schistosoma japonicum / 生物工程学报

Radiation sensitive protein 23 (RAD23) is a nucleotide excision repair (NER) protein that plays an important role in Ubiquitin-proteasome pathway (UPP). Schistosoma japonicum radiation sensitive protein23 (SjRAD23) cDNA sequences were amplified by PCR and cloned into pET28a (+) vector to construct recombinant expression plasmid pET28a(+)-SjRAD23. The recombinant protein was expressed as both inclusion bodies and the supernatant in Escherichia coli BL21 (DE3) cell. Immunofluorescence observation shows that SjRAD23 was mainly distributed on the tegument surface of the worms. ELISA assay reveals that specific IgG, IgG1 and IgG2a antibodies could be detected in the sera of rSjRAD23 immunized mice. Western blotting analysis shows that the recombinant SjRAD23 could be recognized by serum specific to soluble adult worm antigen of S. japonicum. BALB/c mice vaccinated with rSjRAD23 combined with 206 adjuvant revealed 35.94% worm reduction and 40.59% liver egg reduction when compared with that of the adjuvant control

Cloning,expression and immuno-protection analysis of a gene encoding tro-ponin T of Schistosoma japonicum(SjTnT) / 中国血吸虫病防治杂志

Objective To clone cDNA encoding troponin T of Schistosoma japonicum(SjTnT),and evaluate the protective efficacy induced by recombinant SjTnT in BALB/c mice against S. japonicum challenge infection. Methods The SjTnT gene was amplified from 28-day-schistosome cDNAs by PCR and then subcloned into pET28a(+). The recombinant SjTnT protein (rSjTnT)was expressed in Escherichia coli BL21(DE3)cells. The serum specific to rSjTnT was prepared by immunized BALB/c mice with the recombinant antigen,and the immunogenicity of rSjTnT was detected by Western blotting and ELISA. The immuno-protective efficacy induced by rSjTnT in BALB/c mice was evaluated according to the reduction in worm and egg counts. Results The cDNA encoding SjTnT was successfully cloned and expressed in E. coli. Western blotting showed that rSjTnT had a good immunogenicity. The high level of specific IgG antibodies was detected,and 33.89% worm reduction and 43.94% liver egg reduction were obtained in mice vaccinated with rSjTnT combined with Seppic 206 adjuvant compared with those in the adjuvant control group. Conclusions rSjTnT could induce partial immuno-protection against S. japonicum infec-tion in BALB/c mice. This study provided a basic for understanding the biological function of SjTnT.

Characterization and immunoprotective effect of SjIrV1, a 66 kDa calcium-binding protein from Schistosoma japonicum / 生物工程学报

Calcium-binding protein is an indispensable protein which performs extensive and important functions in the growth of Schistosoma japonicum. Based on our primary study on tegument surface proteins of S. japonicun, a cDNA encoding a 66 kDa calcium-binding protein of S. japonicum (Chinese strain) was cloned, sequence analysis revealed that it was identical with that of SjIrV1 of Philippines strains S. japonicum. The expression of SjIrV1 were detected by Real-time PCR, using cDNA templates isolated from 7, 14, 21, 28, 35 and 42 days worms and the results revealed that the gene was expressed in all investigated stages, and the mRNA level of SjIrV1 is much higher in 42 d female worms than that in 42 d male worms. The cDNA containing the open reading frame of IrV1 was subcloned into a pET28a (+) vector and transformed into competent Escherichia coli BL21 for expression. The recombinant protein was purified using a Ni-NTA purification system, and confirmed by high performance liquid chromatography (RP-HPLC) and tandem mass spectrometry (MS/MS). Western blotting analysis showed that recombinant SjIrV1 (rSjIrV1) could be recognized by the S. japonicum infected mouse serum and the mouse serum specific to rSjIrV1, respectively. Immunofluorescence observation exhibited that SjIrV1 was mainly distributed on the tegument of the 35-day adult worms. ELISA test revealed that IgG, IgG1 and IgG2a antibodies are significantly increased in the serum of rSjIrV1 vaccinated mice. The study suggested that rSjIrV1 might play an important role in the development of S. japonicum.

Investigation report of deviation of nasal septum about personnels who take part in physical examination in the enterprise and facilities of Qinhuangdao / 中国基层医药

ObjectiveTo explore the morbidity rate of deviation of nasal septum about personnels who take part in physical examination in the enterprise and facilities of Qinhuangdao,and clinic symptom.MethodsTransverse questionnaire investigation and normal physical examination were adopted.2604 personnels who take part in physical examination were choiced.ResultsMorbidity rate of deviation of nasal septum was 17.6％.The rate of men' s exceeds that of women' s.ConclusionMorbidity rate of deviation of nasal septum was rather high,clinic symptom of snuffle was primary.

Effects of different doses of dexmedetomidine on MACBAR of sevoflurane in patients undergoing lower abdominal surgery / 中华麻醉学杂志

Objective To investigate the effects of different doses of dexmedetomidine on the minimum alveolar concentration of sevoflurane for blunting responses to skin incision ( MACBAR ) in patients undergoing lower abdominal surgery. Methods Sixty ASA Ⅰ or Ⅱ patients, aged 25-55 yr, weighing 45-75 kg, undergoing electire lower abdominal surgery under general anesthesia, were randomly divided into 4 groups ( n = 15 each): control group (Do group) and 3 dexmedetomidine groups (D1, D2 and D3 groups). The patients were unpremedicated.Dexmedetomidine was not used in group D0. A loading dose of dexmedetomidine 0.1μg/kg was injected iv over 10 min, and then dexmedetomidine was infused at a rate of 0.4, 0.8 and 1.2 μg· kg- 1 · h - 1 for 30 min in groups D1-3 respectively. Anesthesia was induced with inhalation of 8 % sevoflurane. Laryngeal mask airway was inserted when BIS value decreased to 45-55. The patients were mechanically ventilated with inhalation of sevoflurane and a mixture of 50% nitrous oxide and 50% oxygen, and the fresh gas flow was set at 1 L/min. In D0-3 groups, the initial end-tidal concentrations of sevoflurane were 3.0%, 2.5%, 2.0% and 1.5% respectively. The patients' response to skin incision was described as effective if MR or MAP increased by ＜ 15%, or ineffective (MR or MAP increased by ≥ 15%). When the response was effective, the end-tidal concentration of sevoflurane was decreased in the next patient, when ineffective, increased, and the ratio between the two successive concentrations was 0.9.The MRCBAR of sevoflurane was determined by up-and-down method, and 95% confidence interval was calculated.Results The MRCBAR (95% confidence interval) of sevoflurane was 2.85% (2.44%-3.32%), 1.91%(1.61%-2.26%), 1.52% (1.31%-1.77%), and 1.34% (1.15%-1.57%)in D0-3 groups respectively. The MRCBAR of sevoflurane was significantly lower in D1-3 groups than in D0 group, and in D2 and D3 groups than in group D1 (P ＜0.05＝. There was no significant difference in MRCBAR of sevoflurane between D2 and D3 groups (P ＞0.05) .ConclusionContinuous infusion of dexmedetomidine at 0.4, 0.8 and 1.2 μg·kg-1 ·h-1 for 30 min results in a decrease in MACBAR of sevoflurane and enhances the inhibitory effect of sevoflurane on the stress response, and in a dose-dependent manner.

Role of mitochondrial ATP-sensitive potassium channels in attenuation of ischemia-reperfusion injury by lidocaine pretreatment in isolated rat heart / 中华麻醉学杂志

Objective To investigate the role of mitochondrial ATP-sensitive potassium(mito-KATP)channels in attenuation of ischemia-reperfusion(I/R)injury by lidocaine pretreatment in the isolated rat heart.Methods Adult female Wistar rats weighing 220-250 g were anesthetized with intraperitoneal 3% pentobarbital 35 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%CO2 at 37 ℃.Twenty-four isolated rat hearts with I/R injury were randomly divided into 3 groups(n = 8 each):group I/R,lidocaine group(group L)and lidocaine + glibenclamide group(group LG).After 10 min of equilibration,group C,L and LG received 20 min of perfusion with K-H solution,K-H solution containing lidocaine 2.5 mg/L and K-H solution containing lidocaine 2.5 mg/L + glibenclamide(a blocker of mito-KATP channels)10 μmol/L,respectively,then subjected to 30 min of ischemia followed by 60 min of reperfusion.HR,left ventricular developed pressure(LVDP),+ dp/dtmax and - dp/dtmax were recorded at the end of equilibration(T0)and at 15,30,45 and 60 min of reperfusion(T1-4).Coronary effluent was collected at T0 and T4 for determination of lactate dehydrogenase(LDH)and creatine kinase(CK)activities.Myocardial tissues were obtained from cardiac apex at T4 for determination of Na+ -K+ -ATPase and SOD activities and MDA and Ca2+ contents.Results Compared with group I/R,HR,LVDP,+ dp/dtmax and - dp/dtmax were significantly increased,CK and LHD activities were decreased,Na+ -K+-ATPase and SOD activities were increased,and MDA and Ca2+ contents were decreased in group L(P ＜0.05).Compared with group L,HR,LVDP,+ dp/dtmax and -dp/dtmax were significantly decreased,CK and LHD activities were increased,Na+ -K+ -ATPase and SOD activities were decreased,and MDA and Ca2+ contents were increased in group LG(P＜0.05).Conclusion The mechanism by which lidocaine pretreatment attenuates I/R injury to the isolated rat heart is related to mito-KATP channel opening.

Cloning, expression and characterization of a gene encoding alpha2 subunit of the proteasome in Schistosoma japonicum / 生物工程学报

The 26S proteasome is a proteolytic complex responsible for the degradation of the vast majority of eukaryotic proteins. Regulated proteolysis by the proteasome is thought to influence cell cycle progression, transcriptional control, and other critical cellular processes. A novel Schistosoma japonicum gene (GenBank Accession No. AY813725) proteasome alpha2 subunit (SjPSMA2) was cloned. Sequence analysis revealed that the ORF of SjPSMA2 gene contains 708 nucleotides encoding 235 amino acids, and the molecular weight was estimated to be 25.84 kDa. Real-time PCR analysis showed that this gene expressed in 7 d, 13 d, 18 d, 23 d, 32 d and 42 d schistosoma. The mRNA level of SjPSMA2 was lower in 7 d and 23 d schistosomulum than that in other stages. The SjPSMA2 cDNA fragment was subcloned into an expression vector pET28a(+) and transformed into E. coli BL21 (DE3) cells. After induction with IPTCQ the 30 kDa fusion protein was produced as included bodies. Western-blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation, and the protein in native could be detected. After immunization of BALB/c mice with the fusion protein, the reduction rates of worm counts and liver egg counts were 12.33% and 35.23%. ELISA results revealed that the vaccinated group showed a significant increase in the level of IgG antibody. This study provided an important basis for investigating the regulation mechanism of the proteasome during the development of Schistosoma japonicum.

Cloning and expressing of cyclophilin B gene from Schistosoma japonnicum and the analysis of immunoprotective effect / 生物工程学报

The present study was intend to clone and express the cDNA encoding Cyclophilin B (CyPB) of Schistosoma japonicum, its preliminary biological function and further immunoprotective effect against schistosome infection in mice. RT-PCR technique was applied to amplify a full-length cDNA encoding protein Cyclophilin B (Sj CyPB) from schistosomula cDNA. The expression profiles of Sj CyPB were determined by Real-time PCR using the template cDNAs isolated from 7, 13, 18, 23, 32 and 42 days parasites. The cDNA containing the Open Reading Frame of CyPB was then subcloned into a pGEX-6P-1 vector and transformed into competent Escherichia coli BL21 for expressing. The recombinant protein was renaturated, purified and its antigenicity were detected by Western blotting, and the immunoprotective effect induced by recombinant Sj CyPB was evaluated in Balb/C mice. The cDNA containing the ORF of Sj CyPB was cloned with the length of 672 base pairs, encoding 223 amino acids. Real-time PCR analysis revealed that the gene had the highest expression in 18-day schistosomula, suggesting that Sj CyPB was schistosomula differentially expressed gene. The recombinant protein showed a good antigenicity detected by Western blotting. Animal experiment indicated that the vaccination of recombinant CyPB protein in mice led to 31.5% worm and 41.01% liver egg burden reduction, respectively, compared with those of the control. A full-length cDNA differentially expressed in schistosomula was obtained. The recombinant Sj CyPB protein could induce partial protection against schistosome infection.

Cloning and expression of Sj423 gene encoding the egg-shell precursor protein of Schistosoma japonicum / 中国人兽共患病学报

The gene fragment encoding the egg-shell precursor protein of Schistosoma japonicum was amplified with RT-PCR by using PCR primer designed according to the 423 bp cDNA fragment of the Philippine strain of S.japonicum, the corresponding mDNA fragment of Chinese strain as template and then the 5' and 3' ends of this gene cDNA were amplified with 5' RACE and 3' RACE by using a series of primers designed according to the result of sequencing. Result of sequence analysis showed that this fragment, named as Sj423, contained a complete open reading frame (ORF) of gene encoding the egg-shell precursor protein of S.japonicum.(Chinese strain). As demonstrated by sequencing analysis. No intron could be detected in this gene fragment. This gene was subsequently expressed in E.coli after cloning into the expression vector pET28c(+). The molecular mass of the expressed product of this gene was 20.9 kDa as revealed by SDS-PAGE analysis, and Western blot analysis showed that the recombinant protein expressed could react well with the rabbit antiserum against the worm antigen of S.japonicum;indicating the good antigenicity of this expressed product.

Screening of T7 phage-display cDNA library from liver of Microtus fortis with extracts of schistosomulum and characterization of the positive clones / 生物工程学报

Microtus fortis is naturally resisitent to Schistosoma japonicum. In order to find schistosome-resistence-related genes of Microtus fortis, a T7 phage-display cDNA library from liver of Microtus fortis was screened with the soluble lysate of schistosomula. The specific phages were enriched 375-fold after 3 rounds of biopanning. Ninety-two positive clones picked at random were sequenced and 19 ESTs including 6 unreported genes were obtained. Compared with the negative phage clone control, five positive clones, No.4 (GenBank Accession No.: EW968294), No.13 (GenBank Accession No.: EW968303), No.14 (GenBank Accession No.: EW968304), No.15 (GenBank Accession No.: EW968305) and No.18 (GenBank Accession No.: EW968308) could induce significantly higher schistosomula mortality rate when co-cultivated with schistosomula. According to the function analysis and the shistosomula-killing effect in vitro, the genes encoding CASP8 and FADD-like apoptosis regulator isoform protein, alpha-2-HS-glycoprotein, M4 protein, R3H domain (binds single-stranded nucleic acids) isoform 2 and 3 previously unreported proteins (No.14, No.15 and No.18) obtained here, were schistosomiasis-resistence-related genes of Microtus fortis.

Cloning, expressing and characterizing of a phosphoglycerate mutase gene of Schistosoma japonncum / 生物工程学报

Phosphoglycerate mutase (PGAM) is a key enzyme in glycolytic pathways. With PCR technique based on an EST identified in our lab, a novel gene named SjPGAM (GenBank Accession No. EU374631) was cloned. Sequence analysis revealed that the ORF of SjPGAM gene contained 753 nucleotides, encoding 250 amino acids, and the molecular weight was about 28.26 kD. Real-time PCR analysis showed that the mRNA level of SjPGAM was much higher in the 14 days and 19 days schistosomula than other stages, suggesting that the gene was a schistosomula stage differential expression gene. The SjPGAM cDNA fragment was subcloned into an expression vector pET-28a (+) and transformed into Escherichia coli BL21 cells. In the presence of IPTG, the 31 kD fusion protein was expressed in included bodies. Western blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation. The study provides important basis for investigating the mechanism of the PGAM in the glycolytic pathways of Schistosoma japonnicum.

Observations on protective immunity in mice induced by recombinant Schistosoma japonicum Sj14FABP through several adjuvant formulations / 中国血吸虫病防治杂志

Objective To test the protective immunity in mice induced by recombinant Schistosoma japonicum Sj14FABP through several adjuvant formulations. Methods The recombinant Schistosoma japonicum Sj14FABP was prepared by expression in E. coli as a GST fusion protein (rSj14/GST) and used to vaccinate outbred Kunming mice by using complete Freund's adjuvant (FCA)/incomplete Freund's adjuvant (FIA), Bacillus Calmette-Guerin (BCG) and the immunostimulating complex (ISCOM) as adjuvant respectively. Results The purified recombinant protein rSj14/GST was immunogenic in mice, and 34.3% and 36.0% worm reduction rates were obtained in outbred Kunming mice immunized intradermally with BCG adjuvant and immunized subcutaneously with ISCOM adjuvant respectively, compared with non-vaccinated control group. However, intramuscularly vaccination with rSj14/GST in FCA/FIA was not protective, although the high level of IgG antibody was induced. Conclusion Both BCG and ISCOM are suitable adjuvants for rSj14/GST.

Screen-Printing Enzyme Electrode-Immobilizing the Enzyme with Silk Fibroin / 分析化学

This paper reports the technique to develop a disposable screen-printing base carbon electrode for the utilization in the study of enzyme electrode. The silk fibroin was applied to immobilize the mushroom extracted protein, which contained abundant polyphenol oxidase, on the surface of base electrode when it was chemically moldified by ferrocene. The voltammetric current of this enzyme electrode responded to the concentration of substrate such as catechol or dopamine. The linear range is from 2.0× 10-8 mol/L to 2.0× 10-2 mol/L, it reaches to a 95%steady value of current within less than 30 seconds, and the RSD equals to 2.7%. The service life of this enzyme electrode is at least 10 days and it will be preserved for a longer period at a humid and cool condition. All of these performances of the enzyme electrode make clear that its commercial development would be very possible.