Objective:To explore the value of FilmArray meningitis/encephalitis(ME)Panel in etiological diagnosis of infection in central nervous system(CNS) in Chinese children.Methods:Cerebrospinal fluid(CSF)obtained through lumbar puncture was collected from 145 patients with suspected CNS infection at Shanghai Children′s Medical Center Affiliated to Shanghai Jiaotong University School of Medicine from March 2019 to November 2019.All specimens were cultured simultaneously, which were detected by FilmArray ME Panel, and the results of cerebrospinal fluid culture and FilmArray ME Panel were compared.Results:Among 145 patients with suspected CNS infection, three samples were found to be positive after cerebrospinal fluid culture, and the positive rate was 2.1%(3/145). For the FilmArray ME Panel, 30 specimens were found to be positive, with a positive rate of 20.7%(30/145), and the difference of positive rate between the two methods was statistically significant( χ2=24.927, P<0.05). Among the samples FilmArray ME Panel tested positive with pathogen, 26 specimens were positive with virus making up 17.9%(26/145)and enterovirus(15.2%)was the primary pathogen.In addition, of the 142 specimens cerebrospinal fluid culture negative, 28 samples were tested positive by the FilmArray ME Panel, accounting for 19.7%(28/142). Conclusion:FilmArray ME Panel has the characteristics with high positive rate and could be time-saving.Meanwhile, FilmArray ME Panel has significant advantage in the detection of virus and improves the positive detection rate of virus.

OBJECTIVETo observe the effects of astragalus membranacaus injection on the activity of the intestinal mucosal mast cells (IMMC) and inflammatory response after hemorrahagic shock-reperfusion in rats.

METHODThirty-two Wistar rats were randomly divided into four groups: normal group, model group, low dosage group, (treated with astragalus membranacaus 10 g kg(-1)) and high dosage group (treated with astragalus membranacaus 20 g kg(-1)). Models of hemorrhage shock for 60 minutes and reperfusion for 90 minutes were created. The animals were administrated 3 mL therapeutic solution before reperfusion. At the end of study, intestinal pathology, ultrastructure of IMMC, and expression of tryptase were observed. The levels of MDA, TNF-a, histamine, and SOD activity of intestinal were detected, and the number of IMMC was counted.

RESULTThe degranulation of IMMC was seen in model group and was attenuated by astragalus membranacaus treatment. Chiu's score of model group was higher than that of the other groups. Astragalus membranacaus could attenuate the up-regulation of the Chiu' s score, the levels of MDA and TNF-alpha, expression of tryptase, and the down-regulation of SOD activity and histamine concentration. The Chiu's score and MDA content were negatively, while SOD activity was positively correlated to the histamine concentration respectively in the four groups.

CONCLUSIONAstragalus membranacaus can reduce small intestine mucosal damage by inhibiting the activity of IMMC after hemorrhage shock reperfusion.

Animals ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Injections, Intravenous ; Intestinal Mucosa ; metabolism ; pathology ; Intestine, Small ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mast Cells ; drug effects ; metabolism ; ultrastructure ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; pathology ; Shock, Hemorrhagic ; metabolism ; pathology ; Tryptases ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To establish a quantitative RT-PCR method with self-quenched fluorogenic probe for detection of bcr/abl mRNA in patients with chronic myeloid leukemia for providing a useful tool for diagnosis of CML,evaluation of therapeutic effect and monitoring of minimal residual disease(MRD). Methods bcr/abl gene from cultured K562 cells was amplified by conventional RT-PCR.The standard quantitative plasmid was constructed by A-T clone method.The self-quenched fluorogenic quantitative RT- PCR method(FQ-RT-PCR)for determination of bcr/abl mRNA was established successfully using the ABI PRISM 7000 PCR Detector.The linear range,sensitivity,stability,and repetitiveness of the method were determined.The marrow samples from 25 CML patients and 3 ALL patients were assessed.Results The sensitivity of the FQ-RT-PCR was 10 copies/?l recombined plasmid,and bcr/abl mRNA can be detected from 1 K562 cell in 10~5 normal cells.The linear range was 10~2-10~9 copies/?l recombined plasmid.The coefficient variation(CV)value was 2.1% in intra-assay and 6.1% in inter-assay.The median ber/abl mRNA expression level was 4.50?10~4 copies/?g RNA [(0.45-89.00)?10~4],5.45?10~4 copies/?g RNA [(2.95-19.30)?10~4 ],13.00?10~4 copies/?g RNA [(4.10-89.00)?10~4] and 2.35?10~4 copies/?g RNA [(0.45-5.12)?10~4] in 25 CML patients,11 patients in the incipient chronic phase,6 patients in blastic crisis,8 patients in chronic period after treatment,respectively.The bcr/abl mRNA level in blastic crisis was significantly higher than that in chronic phase(q= 3.41,P

OBJECTIVETo investigate cardiac function impairment and myocardial injury in rats with intestinal ischemia-reperfusion and the protective effect of cromolyn sodium.

METHODSThirty-two SD rats were randomized into 4 groups (n=8), namely the sham operation group, model group, 50 mg/kg cromolyn sodium group, and 25 mg/kg cromolyn sodium group. Intestinal damage was induced by clamping the superior mesenteric artery for 45 min followed by reperfusion for 60 min. Cromolyn Sodium was administrated intaperitoneally 15 min before reperfusion. The heart rate (HR), left ventricle pressure (LVSP), and the maximal/minimum rate of LVSP (+dp/dt(max), -dp/dt(max)) were sacrificed immediately before ischemia (baseline, T(0)), at 15 min (T(1)), 30 min (T(2)), 45 min (T(3)) of ischemia, and at 3 min (T(4)), 5 min (T(5)), 10 min (T(6)), 15 min (T(7)), 45 min (T(8)), 60 min (T(9)) of reperfusion. At the end of the experiment, the rats were executed and the hearts were immediately removed for observation of the pathological changes and determination of MDA contents and SOD activity.

RESULTSCompared with the baseline T(0), the HR, +dp/dt(max), -dp/dt(max) and the LVSP were decreased significantly at T(8) and T(9) in the model group and the two cromolyn sodium groups (P<0.05). Compared with the sham operation group, these indices were also significantly decreased at T(8) and T(9) in the model group and the two cromolyn sodium groups, but the model group had significantly lower levels for these indices at T(8) and T(9) than the two cromolyn sodium groups (P<0.05). The score of myocardial injury in the model group and the two cromolyn sodium groups were significantly higher than that of group A, and 50 mg/kg cromolyn sodium group had lower score than the model group (P<0.05). The rats in the model group had significantly higher MDA levels than those in the sham operation group and the 50 mg/kg cromolyn sodium group. SOD activities in the model group and 25 mg/kg cromolyn sodium group was lower than that in the sham operation group (P<0.05), but 50 mg/kg cromolyn sodium group had significantly higher SOD activities than the model group (P<0.05).

CONCLUSIONCromolyn sodium can protect the myocardium against intestal ischemia-reperfusion injury and improve the cardiac function.

Animals ; Cardiotonic Agents ; pharmacology ; Cromolyn Sodium ; pharmacology ; Female ; Heart ; drug effects ; physiopathology ; Heart Rate ; drug effects ; Intestines ; blood supply ; Male ; Malondialdehyde ; blood ; metabolism ; Myocardium ; metabolism ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; blood ; prevention & control ; Superoxide Dismutase ; blood ; metabolism ; Time Factors

OBJECTIVETo summarize the experience of lymph node dissection patterns in hand-assisted laparoscopic radical gastrectomy.

METHODSOne hundred and eleven patients with gastric carcinoma between December 2010 and September 2012 were operated by hand-assisted laparoscopic system designed by us. Clinical data were analyzed retrospectively. The lymph nodes were dissected from left to right together with total tumor resection(reverse lymph nodes scavenge pattern), then digestive tract was reconstructed.

RESULTSTotal gastrectomy, distal gastrectomy and proximal gastrectomy were performed in 57, 46 and 8 cases respectively. Combined cholecystectomy and lateral segment of left liver lobe were needed in 4 and 2 patients respectively, and 1 case underwent combined splenectomy and pancreatic body and tail resection. TNM staging of patients in I(, II(, III(A, III(B, and IIII( were 16, 8, 35, 14, and 38, respectively. Histological type was poorly differentiated in 78 cases, moderate differentiation in 26 cases and good differentiation in 7 cases. The incision length was(6.8±0.3) cm, blood loss was(238.4±113.6) ml, operative time was (171.9±23.3) min, number of removed lymph node was 17.2±5.7, hospital stay was (10.1±3.7) d, postoperative complication rate was 9.0%. One case died during perioperative time.

CONCLUSIONSHand-assisted laparoscopic D2 radical gastrectomy(reverse lymph nodes scavenge pattern) can avoid the multiple conversion of open-laparoscopic operation model, and is beneficial to the standardization for surgical procedure.

Gastrectomy ; Humans ; Laparoscopy ; Lymph Node Excision ; Lymph Nodes ; Neoplasm Staging ; Operative Time ; Postoperative Complications ; Retrospective Studies ; Stomach Neoplasms ; pathology ; surgery

OBJECTIVETo identify the mutation of human androgen receptor gene (AR) in a patient with complete androgen insensitivity syndrome (CAIS).

METHODSDNA sequences of 8 exons and their exon/intron boundaries of the AR gene in the patient were amplified by PCR and directly sequenced.

RESULTSDNA sequencing revealed a nonsense mutation in exon 1, resulting in a change of codon 441 GAA (glutamic acid) to a stop codon (TAA).

CONCLUSIONA novel mutation Glu441stop (GAA to TAA) of the androgen receptor gene leading to complete androgen insensitivity syndrome was identified in this study in a Chinese patient. It may help us further understanding the pathogenesis of CAIS.

Adult ; Androgen-Insensitivity Syndrome ; genetics ; Base Sequence ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Polymerase Chain Reaction ; methods ; Receptors, Androgen ; genetics ; Sequence Analysis, DNA ; methods

OBJECTIVETo evaluate the diagnostic utility of Warthin-Starry silver stain, immunohistochemistry and transmission electron microscopy in the detection of human Bartonella henselae infection and pathologic diagnosis of cat scratch disease (CSD).

METHODSThe paraffin-embedded lymph node tissues of 77 histologically-defined cases of cat scratch disease collected during the period from January, 1998 to December, 2008 were retrieved and studied using Warthin-Starry silver stain (WS stain) and mouse monoclonal antibody against Bartonella henselae (BhmAB stain). Five cases rich in bacteria were selected for transmission electron microscopy.

RESULTSUnder electron microscope, the organisms Bartonella henselae appeared polymorphic, round, elliptical, short rod or bacilliform shapes, ranged from 0.489 to 1.110 microm by 0.333 to 0.534 microm and often clustered together. Black short rod-shaped bacilli arranged in chains or clumps were demonstrated in 61.0% (47/77) of CSD by WS stain. The organisms were located outside the cells and lie mainly in the necrotic debris, especially near the nodal capsule. In 72.7% (56/77) of the cases, dot-like, granular as well as few linear positive signals were observed using BhmAB immunostain and showed similar localization. Positive results for both stains were identified in 59.7% (46/77) of the cases. When applying both stains together, Bartonella henselae was observed in 74.0% (57/77) of the case. The difference between the results obtained by WS stain and BhmAB immunostain was of statistical significance (P < 0.05).

CONCLUSIONSBartonella henselae is the causative pathogen of cat scratch disease. WS stain, BhmAB immunostain and transmission electron microscopy are helpful in confirming the histologic diagnosis. Immunostaining using BhmAB can be a better alternative than WS stain in demonstrating the organisms.

Adolescent ; Adult ; Aged ; Antibodies, Bacterial ; blood ; Bartonella henselae ; immunology ; isolation & purification ; ultrastructure ; Cat-Scratch Disease ; diagnosis ; microbiology ; pathology ; Child ; Child, Preschool ; Humans ; Immunohistochemistry ; methods ; Infant ; Lymph Nodes ; pathology ; ultrastructure ; Microscopy, Electron, Transmission ; Middle Aged ; Paraffin Embedding ; Staining and Labeling ; methods ; Young Adult

Buffaloes ; Cells, Cultured ; In Vitro Techniques ; Primary Cell Culture ; Stem Cells ; Tretinoin

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.