Safety of minimally invasive liver resection for resectable hepatocellular carcinoma complica-ted with portal hypertension: a multicenter study / 中华消化外科杂志

Objective:To investigate the safety of minimally invasive liver resection for resectable hepatocellular carcinoma (HCC) complicated with portal hypertension.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopathological data of 807 patients with resectable HCC who underwent minimally invasive liver resection in 8 medical centers, including Sir Run Run Shaw Hospital, Affiliated with the Zhejiang University School of Medicine et al, from June 2011 to November 2022 were collected. There were 670 males and 137 females, aged 58(50,66)years. Of the 807 patients, 173 cases with portal hypertension were divided into the portal hypertension group, and 634 cases without portal hypertension were divided into the non-portal hypertension group. Observation indicators: (1) propensity score matching and comparison of general data of patients between the two groups after matching; (2) intraoperative and post-operative situations; (3) subgroup analysis. Propensity score matching was done by the 1:1 nearest neighbor matching method, with the caliper setting as 0.001. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the rank sum test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was constructed using the non-parameter rank sun test. Results:(1) Propensity score matching and comparison of general data of patients between the two groups after matching. Of the 807 patients, 268 cases were successfully matched, including 134 cases in the portal hypertension group and 134 cases in the non-portal hypertension group. The elimination of the tumor diameter and robot-assisted surgery confounding bias ensured comparability between the two groups after propensity score matching. (2) Intraoperative and postoperative situations. The occlusion time of porta hepatis, cases with intraoperative blood transfusion, cases with postoperative complication, cases with complication >Ⅱ grade of Clavien-Dindo classification, cases of Clavien-Dindo classification as Ⅰ grade, Ⅱ grade, Ⅲ grade, Ⅳ grade, cases with liver related complication were 27.0(15.0,43.0)minutes, 33, 55, 15, 13, 29, 14, 1, 37 in the portal hypertension group, versus 35.0(22.0,60.0)minutes, 17, 25, 5, 14, 9, 4, 1, 13 in the non-portal hypertension group, showing significant differences in the above indicators between the two groups ( Z=-2.15, χ2=6.30, 16.39, 4.38, 20.72, 14.16, P<0.05). (3) Subgroup analysis. Results of subgroups analysis showed that in cases with major live resection, the operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 243.5(174.6,296.3)minutes, 200.0(150.0,600.0)mL, 7.5(6.0,13.0)days in the portal hypertension group, versus 270.0(180.0,314.5)minutes, 200.0 (75.0,450.0)mL, 7.0(5.5,10.0)days in the non-portal hypertension group, showing no significant difference in the above indicators between the two groups ( Z=-0.54, -1.73, -0.92, P>0.05). In cases with non-major live resection, the operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 170.0(120.0,227.5)minutes, 100.0(50.0,200.0)mL, 8.0(5.0,10.0)days in the portal hypertension group, versus 170.0(120.0,227.5)minutes, 100.0(50.0,200.0)mL, 7.0(5.5,9.0)days in the non-portal hypertension group, showing no significant difference in the above indicators between the two groups ( Z=-1.39, -0.10, 1.05, P>0.05). In cases with anatomical liver resection, the operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 210.0(150.0,285.0)minutes, 150.0(50.0,200.0)mL, 8.0(6.0,9.3)days in the portal hypertension group, versus 225.5(146.3,306.8)minutes, 100.0(50.0,250.0)mL, 7.0(6.0,9.0)days in the non-portal hypertension group, showing no significant difference in the above indica-tors between the two groups ( Z=-0.75, -0.26, -0.91, P>0.05). In cases with non-anatomical liver resection, the operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 173.5(120.0,231.5)minutes, 175.0(50.0,300.0)mL, 7.0(5.0,11.0)days in the portal hyper-tension group, versus 186.0(123.0,262.5)minutes, 100.0(50.0,200.0)mL, 7.0(5.0,9.5)days in the non-portal hypertension group, showing no significant difference in the above indicators between the two groups ( Z=-0.97, -1.12, -0.98, P>0.05). Conclusion:Minimally invasive liver resection or even major liver resection is safe and feasible for screened HCC patients complicated with portal hyper-tension, but attention should be paid to the prevention and treatment of postoperative complications.

Effect of 6-Shogaol on invasion and migration of triple negative breast cancer cells via Hedgehog/Glil signaling pathway / 中国药理学通报

Aim To explore the effects of the expression of the transcription faetor Glil of Hedgehog ( Hh ) signaling pathway and the 6-Shogaol mediated Hedge- hog/Glil pathway on the proliferation, invasion and migration in MDA-MB-231 eells of triple negative breast eaneer.Methods MDA-MB-231 eells were transfected by lentiviral vectors to stably overexpressed Glil gene.The overexpression efficiency of Glil was verified by qRT-PCR and Western blot.CCK-8 and EdlJ assays were used to detect the effect of Glil expression and 6-Shogaol on cell viability.Cell scratch assay and Transwell assay were used to detect the ability of migration and invasion.Western blot was used to detect the proteins expression of Hedgehog signaling pathway and other related genes.Results MDA-MB- 231-Glil overexpression cell line was successfully established.When Gli 1 gene was overexpressed, the invasion and migration ability of cells was significantly improved, anrl the expression of Hh signaling pathway gene Glil , EMT marker gene Vimentin, Hippo signaling pathway genes YAP and TEAD4 inereased.When the expression of Glil was inhibited by the Hh/Gli pathway inhibitor Gant61 , the proliferation, invasion and migration abilities were suppressed.When the eells were treated with 6-Shogaol, the abilities of proliferation, invasion and migration were inhibited as well as the proteins expression of Glil , Vimentin, YAP and TEAD4 deereased.Conclusions Glil gene ean promote the invasion and migration of MDA-MB-231 eells.6-Shogaol ean inhibit proliferation, invasion and migration of breast eaneer eells through Hedgehog signaling pathway, suggesting that transcription factor Glil may be one of the targets of 6-Shogaol.

Rapid screening of Salmonella by loop-mediated isothermal amplification combined with lateral flow dipstick / 上海预防医学

Objective:To establish a specific and sensitive method using loop-mediated isothermal amplification for rapid screening of Salmonella. Methods:The invA gene sequence of Salmonella was downloaded from GenBank. After homology comparison with DNAMAN software， amplification primers were designed in the conserved region， and a LAMP-LFD detection method was established. The reaction system was optimized， and the specificity and sensitivity of the method were verified. Results:The sensitivity of this method to detect Salmonella DNA was up to 1.0×101 copies/μL. The positive rate of anal swabs was the same as that of fluorescent PCR. Meanwhile， LAMP-LFD was easy to operate and did not need expensive instruments. The detection result could be obtained within 30 minutes. Conclusion:The LAMP-LFD method established in this study is rapid， simple， sensitive and specific， which is suitable for rapid screening of Salmonella.

Comparison of the effect of small incision cataract surgery whether combined with nucleus chopping or not in the treatment of cataract / 中国基层医药

Objective To compare efficacy of small incision cataract surgery whether combined with nucleus chopping or not in the treatment of cataract. Methods From March 2014 to September 2015,125 cases of age-related cataract(146 eyes) in the the First People′s Hospital of Xiantao were divided into 60 cases(75 eyes) of observation group and 65 cases(71 eyes) of control group by method of random sampling. The observation group accepted small incision cataract surgery combined with chopping nucleus. The control group only underwent small incision cataract surgery. The postoperative visual acuity,corneal astigmatism and operation time,and complications and so on in the two groups were compared. Results There was statistically significant difference in postoperative visual acuity after 1 week between the two groups(Z = -2. 078,P <0. 05),but there was no statistically significant difference in postoperative visual acuity after 1 month or 3 months between the two groups(Z= -0. 960,-0. 743,all P>0. 05). The postoperative corneal astigmatism after 1 week or 1 month between the observation group and the control group had statistically significant differences[(1. 33 ± 0. 45) D vs. (1. 52 ± 0. 49) D,(1. 03 ± 0. 42) D vs. (1. 18 ± 0. 44)D,t=2. 442,2. 108,all P<0. 05)],but there was no statistically significant difference in postoperative corneal astigmatism after 3 months between the two groups(t =0. 432,P >0. 05). There was no statistically significant difference in the operation time between the observation group and the control group[(11. 28 ± 2. 32) min vs. (11. 87 ± 2. 52)min,t=1. 473,P>0. 05]. One case of posterior capsular rupture occurred in the two groups,respec-tively. But serious complications such as lens nucleus escaped into vitreous cavity, explosive haemorrhage from the choroid or corneal endothelial decompensated had not been found. Conclusion The small incision cataract surgery combined with nucleus chopping has advantages in acquiring fast visual rehabilitation acuity in the early stage after operation,lower corneal astigmatism compared to that without nucleus chopping. Therefore,the small incision cataract surgery combined with nucleus chopping is worthy of clinical popularization and application.

Effects of Gemcitabine Combined with Radiotherapy and Chemotherapy on Angiogenesis and Invasion Ability of Cervical Cancer Cell / 中国药房

OBJECTIVE:To investigate the effects of gemcitabine combined with radiotherapy and chemotherapy on angiogen-esis and cell invasion ability of cervical cancer. METHODS:Totally 37 patients undergoing radical operation selected from Chongq-ing Kaizhou District People's Hospital during Jan. 2015-Jun. 2016 were divided into observation group(n=19)and control group (n=18). Control group was given 3-D conformal radiation and chemotherapy which included Cisplatin injection 70 mg/m2,ivgtt+Fluorouracil injection 600 mg/m2,ivgtt on 1st and 21st day after radiotherapy. Observation group was additionally given Gemcitabi-ne hydrochloride for injection 800 mg/m2,ivgtt,1st and 8th day after radiotherapy. A treatment course lasted for 28 d. The radical hysterectomy and pelvic lymphadenectomy were conducted after 2 courses of treatment. Clinical efficacy,microvessel density (MVD) of tumor,vascular endothelial growth factor (VEGF),Wnt1,Wnt3a,Wnt8 and β-catenin level were observed in 2 groups. The relationship of MVD with VEGF was analyzed,and the occurrence of ADR was recorded. RESULTS:The total re-sponse rate of observation group(84.2%)was significantly higher than control group,with statistical significance(P<0.05). The levels of MVD,VEGF,Wnt1,Wnt3a,Wnt8 and β-catenin in 2 groups were significantly higher than healthy tissue;above index-es of focus tissue in observation group were significantly lower than control group,with statistical significance (P<0.05). MVD was positively correlated with VEGF. There was no statistical significance in the incidence of ADR between 2 groups (31.6% vs. 27.8%,P<0.05). CONCLUSIONS:Gemcitabine combined with radiotherapy and chemotherapy can regulate the angiogenesis of cervical cancer cell and the activity of Wnt/β-catenin signal pathway,and reduce the invasion ability of cervical cancer cell,which may be the important mechanism of anti-tumor effect with good safety.

AG490 regulated the invasion and metastasis of gallbladder SGC -996 cells through JAK/STAT3 pathway / 中国基层医药

Objective To study the effect of AG490 on the invasion and metastasis of gallbladder SGC -996 cells regulated by JAK/STAT3,and discuss the related mechanism.Methods Cell viability treated with different concentrations of AG490(50,100,200μmol/L)was detected by MTT method.The ability of invasion and metastasis of gallbladder cells was evaluated by Transwell membrane count.The SGC -996 cell apoptosis was detected by flow cytometry.The protein expression of JAK/STAT3 pathway was detected by Western blot.Results The viability inhi-bition rates of different concentrations of AG490 for SGC -996 cells were (17.49 ±3.41)%,(38.66 ±4.57)%, (79.15 ±6.29)% respectively,and with the increasing of concentration,cell viability decreased obviously.Compared with the control group[(1.39 ±0.21)%],the differences were statistically significant(t =8.162,14.111,21.401, all P <0.01 ).The transfer ability inhibition rate of different concentrations of AG490 for SGC -996 cell were (23.18 ±4.53)%,(51.75 ±6.46)%,(81.32 ±7.13)% respectively,and with the increasing of concentration of AG490,the inhibition rate of invasion and metastasis enhanced.Compared with the control group [(1.46 ± 0.42)%],the differences were statistically significant(t =8.269,13.455,19.366,all P <0.01).The apoptosis rate for SGC -996 cells of different concentrations of AG490 were (13.34 ±4.33)%,(28.16 ±6.23)%,(53.61 ± 8.74)% respectively,and cell apoptosis increased with the increasing of concentration.Compared with control group [(0.97 ±0.52)%],the differences were statistically significant(t =4.913,7.533,10.414,all P <0.01).Different concentrations of AG490 can reduce expression of ZFX,STAT3 and Smad1 protein of JAK/STAT3 pathway of SGC -996 cells,compared with the control group,the differences were statistically significant(tZFX =2.154,3.041,4.185, tSTAT3 =7.348,14.892,17.774 and tSmad1 =3.474,5.241,7.718,all P <0.05).Conclusion AG490 can inhibit the invasion and metastasis of gallbladder SGC -996 cells,and the effect depends on dosage.Its mechanism may be relat-ed to the reduction of cell apoptosis and the protein expression of JAK/STAT3 pathway.

Clinical evaluation of short fiber ribbon combined with resin bonding technology for the treatment of food impaction between posterior teeth / 实用口腔医学杂志

Objective:To evaluate the clinical effects of short fiber ribbon combined with resin bonding technology for the treatment of food impaction between posterior teeth. Methods:98 cases of vertical food impaction between posterior teeth( total of 135 vertical food impaction units) were included. 73 units were treated by short quartz fiber ribbon combined with resin bonding technology( SQFRB) and 63 by resin bonding(RB). 12, 24 and 36 months after restoration, clinical effects were evaluated referring to the Modified United States Public Health Service (USPHS) Criteria, data were statistically analyzed. Results:12, 24 and 36 months after treatment the cure rate of SQFRB was 97. 3%, 97. 3% and 95. 9%, inefficacy rate was 0, 0 and 0;the cure rate of RB was 85. 5%, 82. 2% and 82. 2%, the inefficacy rate was 4. 8%, 11. 3% and 12. 9%, respectively(between groups, P<0. 05). Conclusion:Minimally inva-sive restorations using short fiber ribbon combined with resin bonding technology is effective in the treatment of vertical food impaction between posterior teeth.

Study on Effects and Mechanism of Rapamycin on Invasion and Metastasis of Cervical Cancer HeLa Cells / 中国药房

OBJECTIVE:To study the effects and mechanism of rapamycin on invasion and metastasis of cervical cancer HeLa cell. METHODS:HeLa cells were divided into control group and rapamycin low-dose,medium-dose and high-dose groups (10, 30,100 nmol/L). After treated for 48 h,cell viability was measured by MTT assay,and inhibitory rate was calculated;migration and invasion of cell was tested by Transwell assay. The expression of matrix metalloproteinase 2(MMP-2),MMP-9,Vimentin and E-cadherin,and phosphorylation of protein kinase B (Akt),mammalian target of rapamycin (mTOR) were detected by Western blot. RESULTS:Compared with control group,the inhibition rate of cell viability was increased in rapamycin groups(P<0.01);the number of invasion and metastasis cells decreased(P<0.01);the expression of MMP-2,MMP-9 and Vimentin were decreased (P<0.01 or P<0.05);the expression of E-cadherin was enhanced(P<0.01 or P<0.05);the phosphorylation of Akt and mTOR were reduced (P<0.01). CONCLUSIONS:Rapamycin could inhibit invasion and metastasis of HeLa cell via Akt/mTOR signal pathway.

Reacción en cadena de la polimerasa para la identificación de Salmonella spp. usando el gen invA / Polymerase Chain Reaction (PCR) for identification of Salmonella spp. using the invA gene

Se desarrolló un protocolo de PCR para identificar cepas de Salmonella de diferentes procedencias empleando un set de iniciadores PCR específico a Salmonella, para el reconocimiento de una secuencia de 119 pb del gen invA. Los resultados cumplieron con las máximas de la selectividad: inclusividad, pues todas las 14 cepas de Salmonella ensayadas presentaron señal positiva para el gen, exclusividad ya que ninguna cepa no Salmonella (Shigella, E. coli, E. coli O157:H7) mostró señal positiva, robustez porque en las 14 cepas de Salmonella se obtuvo la señal esperada, en un total de cuatro repeticiones con iguales condiciones de trabajo en cuatro días no consecutivos. Asimismo, fue reproducible, pues los resultados fueron idénticos al modificar las condiciones de reacción. Al comparar la PCR con el cultivo convencional, usando preenriquecimiento no selectivo en agua peptonada y enriquecimiento selectivo en caldo selenito-cistina, se apreció que la PCR fue 100% sensible y especifica, con 100% de inclusividad como de exclusividad.

A PCR protocol was developed to identify strains of Salmonella from different sources using a set of “primers” PCR specific to Salmonella, for the recognition of 119 bp invA gene sequence. The results met the maxims of selectivity: inclusiveness, all strains of Salmonella presented positive signal for the gene, exclusivity since no strain Salmonella (Shigella, E. coli, E. coli O157: H7) showed no signal positive robustness because in the 14 strains of Salmonella expected signal was obtained in a total of four repetitions with equal working conditions in four non-consecutive days. Also it was reproducible, results were identical by modifying the reaction conditions. Comparing PCR with conventional culture, using non-selective pre-enrichment in peptone water and selective enrichment in selenite-cystine broth, it was found that the PCR was 100% sensitive and specific, with 100% of inclusiveness, 100% exclusive and 100% of both positive and negative predictive value.

Physical and chemical properties of acetylated low molecular weight heparin and its antineoplastic effect in vitro / 中国药理学通报

Aims To synthesize acetylated low molecu-lar weight heparin( ALMWH) and to detect its physico-chemical properties and antineoplastic activity. Meth-ods LMWH was prepared by degradation of UFH with sodium periodate oxidation and sodium borohydride re-duction, then the LMWH was acetylated by acetic an-hydride where N, N′, -dicyclohexylcarbodiimide ( DCC ) and 4-dimethylaminopyridine ( DMAP ) were used as catalysts. X-ray diffraction(XRD)and Differ-ential Scanning Calorimetry ( DSC ) of ALMWH were obtained. The antiproliferative activity and anti-inva-sive activity were determined on MDA-MB-231 and MCE-7 human breast cancer cells. Results XRD a-nalysis showed that the LMWH degraded from UFH and ALMWH synthesized by acetylation of LMWH be-longed to amorphous structure, however, their DSC curves were significantly different. Compared with LM-WH, ALMWH had more powerful capacity for binding water and lowering anticoagulant activity, more signifi-cantly ALMWH exhibited stranger anti-proliferative and anti-invasive activity than LMWH, especially when it was used in low concentrations. Conclusion The syn-thesized ALMWH possesses a low anticoagulant activi-ty, certain anti-proliferative, anti-invasive and anti-metastatic activity. This study provides a basic method for screening of antineoplastic drug with low toxicity.

Loop-mediated isothermal amplification assay for the detection of Salmonella spp. in pig feces / 대한수의학회지

Salmonella are causative agents of gastroenteritis and systemic disease in animals. The invA gene was selected as a target sequence of loop-mediated isothermal amplification (LAMP) assay for diagnosis of Salmonella infection. The detection limits for broth dilution, spiked feces and enrichment were 10(4), 10(5) and 10(2) CFUs/mL, respectively. The LAMP assay developed in the present study may be a reliable method for detection of Salmonella spp. in pig feces.

Application of SYBR Green real-time PCR assay for the specific detection of Salmonella spp / 대한수의학회지

The aim of this study was to applicate and evaluate a SYBR Green real-time PCR for the specific detection of Salmonella spp. Specificity of the PCR method was confirmed with 48 Salmonella spp. and 5 non-Salmonella strains using invA gene primer. The average threshold cycle (C(T)) of Salmonella spp. was 11.83 +/- 0.78 while non-Salmonella spp. was 30.86 +/- 1.19. Correlation coefficients of standard curves constructed using C(T) versus copy number of Salmonella Enteritidis ATCC 13076 showed good linearity (R2 = 0.993; slope = 3.563). Minimum level of detection with the method was > 10(2) colony forming units (CFU)/mL. These results suggested that the SYBR Green real-time PCR might be applicable for the specific detection of Salmonella spp. isolates.

Estandarización de una PCR para la detección del gen invA de Salmonella spp. en lechuga / Standardization of a PRC method for the detection of the Salmonella spp. invA gene in lettuce

Salmonella spp. es un patógeno bacteriano muy importante causante de diarreas, que es transmitido tanto por la vía fecal-oral, como por alimentos y agua contaminados. En este trabajo se estandarizó una técnica de PCR en lechuga para la detección del gen invA de Salmonella spp.; dicho gen se relaciona con el proceso de invasión al epitelio intestinal. Con la PCR desarrollada en este trabajo se logró estandarizar un método que permite la amplificación del gen invA con una detección de 10² UFC/25 g. Este método acorta los tiempos de respuesta de los resultados presuntivos y brinda información complementaria al cultivo tradicional del patógeno. El estudio del gen invA establece el potencial patógeno del microorganismo presente en la muestra, lo que puede ser de utilidad para la salud pública.

Salmonella spp. is a very important bacterial pathogen that causes diarrhea and which is transmitted both through the fecal-oral pathway, as by contaminated food and water. In this study we standardized a PCR method in lettuce for the detection of the Salmonella spp. invA gene. This gene is related to the invasion of the intestinal epithelium process. With the PCR method developed in this study we were able to standardize a method which permits the amplification of the invA gene with a 10² CFU/25 g detection. This method shortens the response times of the presumptive results and gives complementary information to the traditional culture of the pathogen. The study of the invA gene establishes the pathogenic potential of the microorganism present in the sample, which can be useful for public health purposes.

Ultrasound-guided minimally invasive biopsy techniques in thyroid surgery / 国际外科学杂志

Objective To approach the guide of ultrasound-guided minimally invasive biopsy technique in diagnosis and treatment of thyroid diseases. Methods One hundred and seventeen cases (a total of 131 nodules, nodules 0. 5-1.5 cm in diameter, with an average diameter of approximately 0. 8 cm) with thyroid nodules , under Ultrasound-guided minimally invasive biopsy techniques from October 2008 to July 2009 were analyzed. Result One hundred and seventeen cases of patients with biopsy derived satisfaction were not ap-parent discomfort and complications after biopsy. Across 59 cases of surgical treatment in patients with biop-sy, and preoperative biopsy and postoperative pathologic is not fully consistent in only two cases, 58 patients without surgical treatment, who is combined with symptoms, palpation, serum thyroxine and color Doppler ultrasound and other laboratory examinations, and regularly review treated with symptomatic treatment, the symptoms, and color Doppler ultrasound imaging were much better under the treatment. 8 cases of patients in 117 cases of minimally invasive biopsy have cervical lymph node biopsy at the same time, biopsy patholo-gy and postoperative results are consistent with postoperative pathologic (7 cases of metastatic carcinoma, 1 case of reactive hyperplasia), accuracy was 100%. Conclusion Uhrasound-guided minimally invasive bi-opsy technique have important guiding for diagnosis and treatment of thyroid diseases, consistent with the high rate of clinical, especially for the preoperative diagnosis of small nodules, to avoid unnecessary over-treatment, can also reduce the misdiagnosis of thyroid cancer. While this technology is simple, safe and ac-curate, which have clinical value in the thyroid surgery.

Further Evaluation of Multiplex PCR for Rapid Detection of Salmonella Typhi / 감염과화학요법

Typhoid fever is a class I legally designated communicable disease in Korea; and if remains as an important public health problem in many developing countries. It takes at least 3-5 days to detect and identify Salmonella Typhi (S. Typhi) by classical diagnostic method. For this reason, multiplex PCR (mPCR) was evaluated in detecting and identifying S. Typhi. In this study, forty-three bacterial strains, which consisted of 42 Salmonella enterica serovars and one Citrobacter freundii. were used to evaluate the promptness of mPCR in detecting and identifying S. Typhi. mPCR was performed with four genes which were known for representing Salmonella spp and/or S. Typhi: invA, fliC-d, viaB and prt. invA and prt gene was amplified in all strains and viaB gene was in only S. Typhi. fliC-d gene was amplified in three serovars: S. Typhi, S. Schwarzengrund and S. Livingstone. After specificity test, mPCR was modified as triplex PCR with three genes (invA, fliC-d, and viaB) and the sensitivity test was performed against S. Typhi-inoculated stool samples. mPCR was able to detect S. Typhi cell suspension of 1x105 cfu/mL. We found that modified multiplex PCR was useful to detect S. Typhi from stool samples within 24h whereas it takes 3-5days to detect by classic diagnosis method.

Transcription and expression characteristics of Leptospira virulence-associated protein InvA / 中华微生物学和免疫学杂志

objective To determine the existence of virulence-associated invA gene in different genospeeies of Leptospira interrogans reference strains in China.and to understand the alterations of invA gene transcription and expression of L.interrogans strain Lai before or after infecting cells.Methods PCR was applied to detect the invA gene of four L.interrogans strains belonging to four different genospecies and L.biflexa strain Patoc Ⅰ.The entire invA genes from the L.interrogans strains were cloned and then sequenced.The prokaryotic expression system of invA gene of L.interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai was constructed.Using Ni-NTA affinity chromatography,the target recombinant protein rInvA was extracted and purified.Rabbits were immunized with rInvA to obtain antiserum and the titer of antiserum was determined by immunodiffusion test.A model of L interrogans strain Lai infecting human embryo kidney cell line HEK293 was established to detect the alterations of invA gene transcription and expression of the leptospiral strain before or after infecting the host cells by real-time fluorescent quantitative RTPCR and western blot assay.Results All the four tested L.interrogans strains had invA gene whereas L.biflexa strain Patoc Ⅰ not.The similarity of nucleotide and putative amino acid sequences of invA genes from the four L.interrogans strains belonging four different genospecies were 99.33%-100%and 98.66%-100%,respectively.The constructed prokaryotic expression system could efficiently express rInvA and the immunodiffusion titer of rabbit anti-rInvA serum was 1:16.After L.interrogans strain Lai infecting HEK293 cells for 30 min or above,the microbe could adhere the surface of the cells.On the 30 min after the infection,the mRNA level of invA gene of L.interrogans strain Lai was remarkably upregulated,and on the 45 min after infection the mRNA level presented a peak value and then graduated decreased.On the 45 min or 60 min after L.interrogans strain Lai infecting HEK293 cells,InvA protein could be detectable but before infection or after infection for over 90 min the InvA protein expression was negative.Conclusion The invA gene is a unique gene for pathogenic L.interrogans.The invA gene of L interrogans has characteristics of cell-touched expression and transient expression,which may have a close correlation with adhering and invading host cells.

Supracondylar and intercondylar femur comminuted fractures treated percutaneously with the GSH re-strograde femural nail / 中华创伤骨科杂志

Objective To find an ideal operative method of i nteral fixation for supracondylar a nd in-tercondylar femur comminuted fractures.Methods 30cases with supracondylar and inte rcondylar femur frac-tures were treated percutaneously with retrograde femoral intramedullary nail(GSH)in condition of close reduction technique with assistance of endosc ope and fluroscope.Results The mean 115ml blood loss was seen in 30cases without grafting intraoperatively.Thigh edema recovery and over90?knee flexion motion were seen one week after the operation.All fractu res were united in 6months.Postoperative following-up for 18months revealed the mean 125?knee motion,normal gait in all cases,no varus or valgus defo rmities and no osteoarthotitis.Conclusion The percutaneous tech.of retrograd e nailing with assistance of endosco pe and fluroscope is an ideal surgery method to treat supracondylar and intercondylar femur factures.

Histopathologic Findings After Experimental Penetrating Keratoplasty Using Expanded Polytetrafluoroethylene(Gore-Tex(R))in Rabbits: Treatment of Corneal Perforation Using Synthetic Materials. Report I

Although there had been many studies for evaluating an usefulness of expanded polytetrafluoroethylene(Gore-Tex(R)) as a temporary treatment of corneal perforation and a supporting skirt of keratoprosthesis, complications and instability of graft had limited the clinical applications. To find histopathologic factors that can contribute to the instability of graft and to try to modify the surgical methods with models of graft, serial examinations including biomicroscopy and histopathology were performed. According to the results, we were able to divide the causes of instability into two categories as a function of time. Epithelial down growth into the interface between graft and cornea with absence of epithelial coverage over the graft may cause leakage from anterior chamber in early period. Absence of fibrovascular invasion into Gore-Tex(R) resulting no cohesion between graft and cornea might be a factor of instability in late period. Based on these results, modification of surgical techniique and models of graft are needed. Several methods of modificiation are suggested in this report.