OBJECTIVEThis paper aims to develop a monoclonal antibodies (MAbs)- based ELISA for detecting Chlamydophila pneumoniae (C. pneumoniae) antigens in humans with the variable domains (VD) 2 and 3 of the major outer membrane protein (MOMPVD2-VD3) and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae.

METHODSMOMPVD2-VD3 were overexpressed in Escherichia coli and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I (156 patients with typical respiratory illness clinically confirmed before) and Group II (57 healthy donors).

RESULTSIn Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II "healthy donors", 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively.

CONCLUSIONThe novel MOMPVD2-VD3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.

Animals ; Antibodies, Monoclonal ; Bacterial Outer Membrane Proteins ; immunology ; Chlamydophila Infections ; diagnosis ; microbiology ; Chlamydophila pneumoniae ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Mice ; Protein Structure, Tertiary

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.

OBJECTIVETo investigate the diagnostic and therapeutic effect of bronchofiberscopy in the management of severe thoracic trauma.

METHODSA retrospective study was conducted on 207 consecutive patients with severe thoracic trauma enrolled in our hospital between January 2008 and June 2012. During the period, 488 bronchofiberscopies and lavages were done. The bronchofiberscope was inserted through tracheal incision (282), nasal cavity (149) and oral cavity (57). Intensive SaO2 monitoring as well as blood gas analysis were performed pre-, intra- and postoperatively. Simultaneously oxygen therapy or ventilatory support was given. Sputum culture was done intraoperatively.

RESULTSDiagnosis in 207 cases was confirmed by bronchofiberscopy. The result of sputum culture was positive in 78 cases. Lavage was performed on 156 cases. SaO2 significantly increased after bronchofiberscopies as well as lavages and PaO2 obviously improved 2 h after surgery (both P less than 0.05). Heart rate and respiratory rate decreased. There was no bronchofiberscopy-related death.

CONCLUSIONBronchofiberscopy plays an important role in the diagnosis and treatment of severe thoracic trauma, which can not only timely diagnose bronchial injury and collect deep tracheal sputum for bacterial culture but also effectively remove foreign body, secretion, blood and sputum crust in the airway, manage obstructive atelectasis and pneumonia, and significantly improve respiratory function and treatment outcome.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bronchoalveolar Lavage ; Bronchoscopy ; Female ; Fiber Optic Technology ; Humans ; Injury Severity Score ; Male ; Middle Aged ; Retrospective Studies ; Thoracic Injuries ; diagnosis ; etiology ; surgery ; Treatment Outcome

PURPOSETraumatic diaphragmatic rupture (TDR) needs early diagnosis and operation. However, the early diagnosis is usually difficult, especially in the patients without diaphragmatic hernia. The objective of this study was to explore the early diagnosis and treatment of TDR.

METHODSData of 256 patients with TDR treated in our department between 1994 and 2013 were analyzed retrospectively regarding to the diagnostic methods, percentage of preoperative judgment, incidence of diaphragmatic hernia, surgical procedures and outcome, etc. Two groups were set up according to the mechanism of injury (blunt or penetrating).

RESULTSOf 256 patients with a mean age of 32.4 years (9-84), 218 were male. The average ISS was 26.9 (13-66); and shock rate was 62.9%. There were 104 blunt injuries and 152 penetrating injuries. Preoperatively diagnostic rate was 90.4% in blunt injuries and 80.3% in penetrating, respectively, P < 0.05. The incidence of diaphragmatic hernia was 94.2% in blunt and 15.1% in penetrating respectively, P < 0.005. Thoracotomy was performed in 62 cases, laparotomy in 153, thoracotomy plus laparotomy in 29, and combined thoraco-laparotomy in 12. Overall mortality rate was 12.5% with the average ISS of 41.8; and it was 21.2% in blunt injuries and 6.6% in penetrating, respectively, P < 0.005. The main causes of death were hemorrhage and sepsis.

CONCLUSIONSDiagnosis of blunt TDR can be easily obtained by radiograph or helical CT scan signs of diaphragmatic hernia. For penetrating TDR without hernia, "offside sign" is helpful as initial assessment. CT scan with coronal/sagittal reconstruction is an accurate technique for diagnosis. All TDR require operation. Penetrating injury has a relatively better prognosis.

Abdominal Injuries ; diagnostic imaging ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Diaphragm ; diagnostic imaging ; injuries ; Female ; Humans ; Male ; Middle Aged ; Multiple Trauma ; diagnostic imaging ; Retrospective Studies ; Rupture ; Thoracic Injuries ; diagnostic imaging ; Tomography, X-Ray Computed ; Wounds, Nonpenetrating ; diagnostic imaging ; Wounds, Penetrating ; diagnostic imaging

OBJECTIVETo clone the plasmid protein pORF8 of Chlamydia trachomatis and localize its expression in Chlamydia-infected cells.

METHODSpORF8 gene was amplified and cloned into pGEX-6p vector, and the pORF8 fusion protein was expressed in E.coli XL1 Blue. After purification with glutathione-conjugated agarose beads, the pORF8 fusion protein was used to immunize BALB/c mice to generate polyclonal antibodies against pORF8 protein. The antibodies obtained were used to localize the plasmid protein pORF8 in Chlamydia-infected cells with immunofluorescence assay (IFA).

RESULTSThe pORF8 gene 744 bp in length was successfully cloned and the GST fusion protein with a relative molecular mass of 54 000 was obtained. The cellular distribution pattern of the plasmid protein pORF8 was similar to that of the major outer membrane protein (MOMP), a known C. trachomatis inclusion body protein, but not to that of chlamydial protease-like activity factor (CPAF, a secreted protein).

CONCLUSIONThe plasmid protein pORF8 is localized on the bacterial organism as an inclusion body protein in C. trachomatis-infected cells. The cellular location of pORF8 protein can potentially provide important insights into the pathogenesis of C. trachomatis.

Animals ; Antibodies ; immunology ; Bacterial Proteins ; biosynthesis ; genetics ; Chlamydia Infections ; metabolism ; Chlamydia trachomatis ; chemistry ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HeLa Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Plasmids ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

Objective To explore whether aggressive treatment of primary focus can benefit nonsmall cell lung cancer(NSCLC)patients with synchronous brain metastasis,and search the appropriate treatment protocols.Methods The clinical data of 19 NSCLC patients with synchronous brain metastasis,received treatment at our Cancer Center from January 2000 to January 2009,were reviewed; their treatments and survival statuses were analyzed.Results Median survival time of these patients was 14.5 months; the 1-year survival rate was 52.6％,and 2-year survival rate was 17.5％.Patients had different survival rates when different treatments were given to the primary focus,and significant difference was noted(x2=10.532,P=0.005); after neurosurgical intervention,patients underwent thoracic operation and chemotherapy(24.9 months)had a significantly longer survival time than those underwent chemotherapy(14.5 months)or palliative therapy(5.4 months,P＜0.05).The survival time of patients with single metastases was 16.3 months,and that of those with multiple metastases was 5.4 months; and significant difference was noted between them(P＜0.05).Conclusion Aggressive therapy including neurosurgical intervention,pulmonary resection and chemotherapy should be recommended for NSCLC patients with synchronous brain metastasis,especially those with single brain metastasis.

OBJECTIVETo evaluate the value of 16-slice spiral CT in the demonstration of coronary artery and in the diagnose of coronary artery stenosis.

METHODSPlain and enhanced CT scans were performed with a 16-slice CT scanner (Sensation 16, Siemens, Germany) in 230 patients with suspected coronary heart disease (CHD). Parameters of the plain scan were: 120 kV, 133 mA, slice collimation 16 mm x 1.5 mm, rotation time 0.42 seconds, increment 1.5 mm, and slice width 3 mm. Parameters of the enhanced scan were: 120 kV, 500 mA, slice collimation 16 mm x 0.75 mm, rotation time 0.42 seconds, increment 0.5 mm, and slice width 1 mm. Enhanced CT scan was performed with a rapid intravenous injection of 100 mL iothalamate meglumine (Ultravist) (370 mgI/mL) or Omnipaque (350 mgI/mL) and 30 mL 0.9% NaCl chaser bolus at a flow rate of 3.5 mL/s. Calcium scoring with plain scan images and two and three dimensional reconstruction with enhanced scan images were made in all cases, among which 30 cases underwent conventional coronary angiography. Demonstration of coronary arteries and their stenosis were evaluated and the factors that might influence the image quality were analyzed.

RESULTSCoronary calcium scores were calculated and coronary artery was demonstrated in our study. In the evaluation of image quality with volume rendering technique (VRT) images, 78.3% of the images were of the first class, 12.2% the second class, and 9.6% the third class. Multi-planar reconstruction (MPR) and maximal intensity projection (MIP) were better than VRT in the demonstration of small branches. The image quality was related to the heart rate, with or without arrhythmia, and breath-hold ability of patients. Comparative study of the stenosis of coronary arteries in 30 cases showed that the sensitivity and specificity of 16-slice coronary CT angiography (CTA) to diagnose significant stenosis were 95.8% and 94.8% respectively.

CONCLUSIONAs a non-invasive and quick method, 16-slice coronary CTA is sensitive and specific to diagnose the stenosis of coronary arteries and can be used as a screening method in the diagnosis of CHD.

Coronary Angiography ; Coronary Disease ; diagnostic imaging ; Coronary Stenosis ; diagnostic imaging ; Female ; Heart Rate ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Tomography, Spiral Computed ; methods

Liver disease is one of the major problems affecting human health. Ultrasound plays an important role in diagnosis and treatment of diffuse and focal liver diseases. However, conventional ultrasound evaluation is subjective and provides limited information. Artificial intelligence (AI) technology may supplement the disadvantages of conventional ultrasound and has been widely used in the field of ultrasound in liver diseases. To date, remarkable progress has been achieved for the use of AI technology in the diagnosis, assessment of therapeutic efficacy and prognosis prediction of liver diseases. This paper reviews the research progress of ultrasound image-based AI technology in the diagnosis and treatment of diffuse and focal liver diseases.

Objective: To investigate the correlation between CLOCK and BMAL1 genes and MEN2 medullary thyroid carcinoma (MTC). Methods: Thirteen cases with MEN2 MTC and thirteen cases with non-MEN2 MTC were selected who were treated in the Yantai Yuhuangding Hospital between January 2013 and September 2021. Clinical indicators such as blood calcitonin level, tumor diameter and metastatic lymph node of patients were collected. The expression differences of CLOCK and BMAL1 between MEN2 MTC and para-carcinoma tissue as well as between MEN2 MTC and non-MEN2 MTC were detected by immunohistochemistry and qPCR. The correlation between lymph node metastasis and CLOCK or BMAL1 expression was analyzed. Protein-protein interaction (PPI) network analysis combined with qPCR and correlation analysis was used to explore the expression regulation relationship between RET and circadian clock genes. The rhythm disorder of MEN2 cells was verified by lipopolysaccharide cell stimulation experiment after dexamethasone rhythm synchronization. Results: MEN2 MTC exhibited typical RET gene mutation. The mean blood calcitonin level, the tumor diameter and the number of metastatic lymph nodes of patients with MEN2 MTC were higher than those of patients with non-MEN2 MTC (t value was 2.76, 2.53, 2.26, all P<0.05). Immunohistochemical results showed that the expression levels of CLOCK and BMAL1 in MEN2 MTC were higher than those in non-MEN2 MTC, while negatively expressed in para-cancerous thyroid follicle. qPCR displayed that the expression of CLOCK gene in cancer tissues was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.68 and 2.86, all P<0.05); the expression of BMAL1 gene in MEN2 MTC was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.21 and 2.35, all P<0.05). Correlation analysis showed that the expression levels of CLOCK and BMAL1 genes were positively correlated with the number of lymph node metastases in patients with MEN2 MTC (r=0.65, P<0.001; r=0.52, P=0.005). PPI network analysis indicated that the expression of CLOCK gene was positively correlated with the abnormal expression of RET gene (r=0.96, P<0.001). With lipopolysaccharide to stimulate cultured cells in vitro after dexamethasone rhythm synchronization, the expressions of CLOCK and BMAL1 in MEN2 MTC cells (0.47±0.22 and 2.60±1.48) at 12 hours of synchronization were significantly lower than those in para-cancerous tissues (1.70±1.62 and 8.23±2.52), the difference was statistically significant(t=5.04, P=0.007; t=3.34, P=0.029). Conclusion: CLOCK and BMAL1 are correlated with the occurrence and development of MEN2 MTC, and may be potential targets for the development of new therapeutic strategies for MEN2 MTC.
ARNTL Transcription Factors/genetics* ; CLOCK Proteins/genetics* ; Calcitonin ; Carcinoma, Neuroendocrine/genetics* ; Dexamethasone ; Humans ; Lipopolysaccharides ; Lymphatic Metastasis ; Multiple Endocrine Neoplasia Type 2a/genetics* ; Thyroid Neoplasms/surgery*

Objective To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD). Methods The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA). Results The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P < 0.05); however, the levels of plasma adrenaline in two groups had no statistical differences (P > 0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05). Conclusions As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.