β-lactam antibiotic binding sites in Streptococcus pneumoniae StkP kinase extracellular region / 中华微生物学和免疫学杂志

Objective:To analyze the binding ability of motifs in the serine/threonine kinase StkP extracellular region (EC-StkP) of Streptococcus pneumoniae to β-lactam antibiotics. Methods:Three motifs (SXXK) in the EC-StkP were mutated into AXXA, respectively or simultaneously. Four mutant plasmids (EC- stkp-AXXA1, EC- stkp-AXXA2, EC- stkp-AXXA3 and EC- stkp-AXXA4) were transfected into recipient cells for cloning and expression. SDS-PAGE combined with gel image analysis was used to detect the expression of the recombinant mutant proteins (EC-rStkP-AXXA1, EC-rStkP-AXXA2, EC-rStkP-AXXA3 and EC-rStkP-AXXA4). The expressed mutated proteins were extracted and purified by Ni-NTA affinity chromatography. The binding abilities of the mutant proteins to penicillin (PCN) and cefotaxime (CTX) were detected by isothermal titration calorimetry (ITC 200) and surface plasmon resonance (Biacore t200). Results:PCN and CTX could not bind to the expressed proteins with mutations in the first or the third motif (EC-rStkP-AXXA1, EC-rStkP-AXXA3, EC-rStkP-AXXA4). EC-rStkP-AXXA2 could weakly bind to CTX, but not to PCN.Conclusions:All three motifs in the EC-StkP of Streptococcus pneumoniae could bind to β-lactam antibiotics with the first and the third motifs being more important.

Experience on postoperative complications of laparoscopic pancreaticoduodenectomy / 中华外科杂志

Objective@#To summarize the incidence and characteristics of postoperative complications after laparoscopic pancreaticoduodenectomy(LPD), and to share our experience on management of complications.@*Methods@#The clinical data of 320 LPD performed by a single team in Sir Run Run Shaw Hospital and Zhejiang Provincial People′s Hospital between September 2012 and September 2017 were retrospectively analyzed, among which there were 196 males and 124 females with age of (60.2±11.6) years old.There were 306 patients who underwent standard LPD, and 14 patients who underwent extended LPD. The patients were divided into 2 groups of former 160 LPD and later 160 LPD according to the time order. By analyzing the differences of clinical outcomes between the two groups, especially focusing on the incidence of postoperative complications.The experience on management of complications was concluded. The prior surgical history of latter group was significantly higher than the former group(30.0%(48/160)vs. 18.8%(30/160), χ2=5.49, P=0.019), and the rest of baseline characteristics remained the comparable (P>0.05). For resectable lesions, LPD was performed by "No back" approach, following the principle of "From distal to cephalad, from ventral to dorsal, and from left to right" . As for the borderline resectable patients, LPD was performed by "Easy first" strategy. Student t test, χ2 test or Fisher test was used to analyzed the data between the two groups respectively.@*Results@#Of 320 LPD patients, 306 cases underwent standard LPD, 14 cases underwent LPD with resection of other organs.There were 278 LPD cases who followed "No back" approach, and 42 cases who followed "Easy first" strategy because of difficulty in creating the retro-pancreatic tunnel. And the overall morbidity was 32.2%(103/320) with reoperation rate of 5.3%(17/320). The perioperative mortality was 0.6%(2/320). The operation time of latter group was ((346.6±48.8)minutes), which was shorter than that of former group((358.0±54.4)minutes)(t=1.97, P=0.048). The blood loss of former and latter group remained comparable((207.9±135.8)ml vs.(189.6±121.4)ml, P=0.205). However, in subgroup analysis, the patients with blood loss less <200 ml of latter group decreased significantly from 59.4%(95/160) to 47.5%(76/160)(χ2=4.53, P=0.033). The overall morbidity of latter group was 28.8%(46/160), indicated a decrease from 35.6%(57/160) of former group without significant difference(P=0.188). Moreover, Grade A/B/C pancreatic fistula rate, Grade A/C bile leakage rate, Grade B/C postoperative hemorrhage rate of the later group tended to decrease, although they also didn′t reach a significant difference. However, the abdominal infection rate decreased significantly(χ2=3.93, P=0.047). The length of hospital stay remained comparable(P=0.156).@*Conclusions@#The most common complications after LPD were postoperative hemorrhage and pancreatic fistula. With specialized team and accumulated experience, the morbidity can decrease progressively by analyzing the leading cause and improving the technical skills.

Cloning and bioinformatics analysis of Giardia lamblia C2 strain SENP gene and prokaryotic expression of SENP catalytic domain / 中国人兽共患病学报

SUMOylation is a post‐translational modification involved in various cellular processes .SUMO‐specific protease (SENP) regulates SUMOylation by removing SUMO from conjugated substrates (deSUMOylation) and promoting maturation of SUMO precursor .In order to express Giardia lambia (C2 strain) SENP catalytic domain in E .coli ,the full‐length open reading frame of SENP was amplified by PCR from Giardia lamblia genome DNA .The PCR product about 1 620 bp was cloned into cloning vector pGM‐T .Sequencing result showed the sequence of SENP in C2 strain was identical with that in Gi‐ardia WB strain .Bioinformatics analysis showed that SENP protein possessed a 372 aa discontinuous ULP catalytic domain at C‐terminal .The catalytic domain of SENP was cloned into prokaryotic expression vector pET‐28a(+ ) .The recombinant vector pET‐28a(+ )‐SENPc was transformed into E .coli Rosetta(DE3) ,then the recombinant SENPc protein was expressed by IPTG induction .SDS‐PAGE and Western blot using anti‐His Tag antibody showed that the expression product of SENPc was a fusion protein with a molecular weight of 43 kD .The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SENP protein provide basis for further functional study of SENP .

MRV comparison of the angle between the right hepatic vein and the inferior vena cava for patients with membranous obstruction of the inferior vena cava / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To determine whether there are differences in both the right hepatic vein (RHV) morphology and the size of the angle between the inferior vena cava and the RHV in patients with membranous obstruction of the inferior vena cava (MOVC),in healthy individuals and in patients with cinhosis (HLC), in order to help guide development of an effective interventional treatment program.</p><p><b>METHODS</b>Consecutive patients (n=248) were divided into the following three groups: group A (control; n=94), group B (MOVC patients; n=68), group C (HLC patients; n=86). The angle between the hepatic vein and inferior vena cava was measured and defined as the T value. The morphology of the RHV was classified as N, U, or I. The difference of the constituent ratio was compared among the three groups for the T value and the angle type.Measurement data was calculated as x ± s,and groups were compared using one-way ANOVA; count data was calculated as relative number, and groups were compared using the chi-square test.</p><p><b>RESULTS</b>The average T value of group B was significantly higher than that of group A (56.1 ± 13.7 vs. 49.3 ± 7.8, P=0.010) and of group C (vs. 51.5 ± 10.0, P < 0.001); the difference was statistically significant (F=8.750, P < 0.001), but there was no significant difference between the groups A and C.N-type proportion of B group was 48.5% (33/68), greater than that of group A(16.0%,15/94) and C (16.3%, 14/86), x² = 20.1, x² =18.6.U-type proportion of B group was 11.8% (8/68), smaller than that of groups A (28.7%,27/94) and C (37.2%, 32/86), 2 2 = 6.70, x² =12.8, and the differences were statistically significant (P < 0.01). For groups A and C, the N and U types were not significantly different.</p><p><b>CONCLUSION</b>The angle between the RHV and the inferior vena cava in MOVC patients is morphologically different from that in healthy humans, with the angle value in MOVC patients being slightly larger. However, this difference is irrelevant to cirrhosis.</p>

A correlation study between high resolution CT appearances and expression of transforming growth factor-β,tumor necrosis factor-α in radiation-induced lung injury of rats / 中华放射学杂志

Objective To study the correlation between high resolution computed tomography manifestations and expression of transforming growth factor beta,tumor necrosis factor alpha in radiationinduced lung injury of rats,and to investigate the vslues of cytokine detection and HRCT scanning for the prediction and early diagnosis of radiation-induced lung injury.Methods Forty-eight Sprague-Dawley (SD)rats were randomly divided into eight groups,group A was normal control group.and group BH were irradiated with a single dose of 15 Gy to the lungs.HRCT scanning Wag performed before and 1 week,2,4,8,12,16,24 weeks after radiation in group A-H respectively.The expression of TGF-beta and TNF-alpha were detected with ELISA. All the rats were killed to observe pathological changes of their lungs.HRCT signs,levels of cytokine were simultaneously compared and analyzed.The t-test and Spenrman rank correlation were used for the statistics.Results Four HRCT signs were observed during the 24 weeks after radiation,including ground-glass opacity(1 case),patchy consolidation(8 cases),massive consolidation(7 cases)and fibrosis(3 cases).The average levels of TGF-beta in group B-H[(3.33±0.47),(3.20±0.65),(3.12±0.45),(3.54±0.80),(3.30±1.13),(2.49±0.67),(4.19±0.22)μg/L,respectively]were hisher than the control group[(0.45±0.14)μg/L.P＜0.05].At 24 weeks the average level increased to the highest peak[(4.19 4-0.22)μg/L,P＜0.05].At 1 week and 2,4,8,12 weeks after radiation,the average level of TNF-alpha in radiation group[(236.52±29.01),(214.91±34.53),(270.97±42.12),(208.83±86.51),(208.83±82.23)ng/L]was hisher than the control group[(31.78±0.92)ng/L,P＜0.05].The average level increased to the highest peak [(270.97±42.12)ng/L,P＜0.05]at 4 weeks.The average level at 16 and 24 weeks[(60.63±38.49),(32.07±1.05)ng/L]dropped to the level similar to the control group(P＞0.05).There were no rank correlations between HRCT manifestations and expression of TGF-beta and TNF-alpha(rs=0.5570 and 0.1013.P＞0.05).HRCT signs were correlated with pathological changes.Conciusions The monitoring of TGF-beta and TNF.alpha in the serum after irradiation can predict the development of radiation-induced lung injury.There are no rank correlations between HRCT manifestations and expression of TGF-beta and TNF-alpha.

Application of Multi-slice Spiral CT Angiography in Thoracic Vascular Emergency / 实用放射学杂志

Objective To evaluate the value of multi-slice spiral CT (MSCT) angiography in diagnosis of thoracic vascularemergency.Methods 25 cases suspected of thoracic vascular emergency were examined with MSCT. Two dimensional and three dimensionalimaging methods were used. Results Central pulmonary embolism 10 cases, aortic dissection 12 cases,aneurysm 2 cases and takayasuarteritis with stricture of kidney artery were found by two dimensional imaging(MPVR,CPR),and the extent, size and details of lesions couldbe showed well. The space relationship and calcifation of aorta were displayed in three dimensional imaging(SSD,VR,MIP). Therelationship between intraluminal embolism and the wall of pulmonary artery, as well as intimal flap and reentry of dissection could bevealuated by CTVE.Conclusion Associated with imaging post processing,MSCT angiography is an effective method in diagnosis of thoracic vascular emergency.