Detection performance of PCR for Legionella pneumophila in environmental samples: a systematic review and meta-analysis.

BACKGROUND: Legionellosis remains a public health problem. The most common diagnostic method to detect Legionella pneumophila (L. pneumophila) is culture. Polymerase chain reaction (PCR) is a fast and accurate method for this detection in environmental samples. METHODS: Four databases were searched for studies that evaluated the detection efficiency of PCR in L. pneumophila. The quality evaluation was conducted using Review Manager 5.3. We used Meta-DiSc 1.4 software and the Stata 15.0 software to create forest plots, a meta-regression, a bivariate boxplot and a Deeks' funnel plot. RESULTS: A total of 18 four-fold tables from 16 studies were analysed. The overall pooled sensitivity and specificity of PCR was 94% and 72%, respectively. The positive likelihood ratio (RLR) and negative likelihood ratio (NLR) was 2.73 and 0.12, respectively. The result of the diagnostic odds ratio (DOR) was 22.85 and the area under the curve (AUC) was 0.7884. CONCLUSION: Establishing a laboratory diagnostic tool for L. pneumophila detection is important for epidemiological studies. In this work, PCR demonstrated a promising diagnostic accuracy for L. pneumophila.

The effect of environmental factors on the differential expression of miRNAs in patients with chronic obstructive pulmonary disease: a pilot clinical study.

Objective: The objective of the study was to analyze the effect of environmental factors on the differential expression of microRNAs in the peripheral blood of migratory and local patients in northern People's Republic of China and on clinical symptoms of local patients in northern People's Republic of China with COPD. Methods: A total of 118 patients in the northern region and 8 migratory patients were enrolled in this prospective study. We collected general information. Blood samples were collected from 9 patients in the Beijing group, from 8 patients in the migratory group and from 9 healthy control subjects. After extracting the total RNA from these 3 groups, serum miRNA was identified by Solexa sequencing. We collected COPD assessment test (CAT) and Modified British Medical Research Council (mMRC) scores at different levels of air pollution and also collected the number of exacerbations over the year prior to the baseline and in the year preceding the follow-up. Results: In total 9 miRNAs were differentially expressed. When air quality index (AQI) >100, the CAT and mMRC scores at baseline were significantly higher than those when the AQI ≤100 (P<0.001). When AQI >100, the follow-up CAT and mMRC scores were significantly higher than those when AQI ≤100 (P<0.001). Follow-up mMRC scores were significantly higher than baseline scores (P=0.04). When AQI ≤100, the baseline CAT score of the group with fewer symptoms was 6.50 (4.00-8.75). However, when AQI >100, the baseline CAT score of this fewer symptoms group was 10.00 (6.25-12.00). The median CAT score was close to 10. When AQI ≤100, the follow-up CAT score of the fewer symptoms group was 8.00 (4.25-12.00). However, when AQI >100, the follow-up CAT score of the fewer symptoms group was 9.50 (6.00-16.75). The median CAT score was close to 10. Conclusion: Environmental factors may cause differential expression of miRNAs in the peripheral blood of migratory and local patients in northern People's Republic of China. Air pollution may aggravate clinical symptoms of patients with COPD.

Pulmonary function tests findings and their diagnostic value in patients with IgG4-related disease.

BACKGROUND: IgG4-related disease (IgG4-RD) is an immune-mediated fibroinflammatory disorder that can affect most organs. To date, there have been no detailed assessments of pulmonary function in patients with IgG4-RD. In this study, we investigated pulmonary function in IgG4-RD patients and evaluated the value of pulmonary function tests (PFTs) in diagnosing IgG4-related respiratory disease (IgG4-RRD). METHODS: This was a retrospective study of 17 patients with IgG4-RD. The patients were divided into two groups: IgG4-RRD group and IgG4-related disease extrapulmonary involvement (IgG4-RDEI) group. The PFT results were compared between the two groups. RESULTS: All patients in the IgG4-RRD group had pulmonary dysfunction. Five of 8 (62.5%) patients in the IgG4-RDEI group had pulmonary dysfunction, despite having normal thoracic computed tomography scans and no respiratory symptoms. Patients in both groups showed restrictive ventilatory dysfunction and abnormal diffusing capacity, and two patients in the IgG4-RRD group had obstructive ventilatory dysfunction. The incidence of diffusing capacity of the lung for carbon monoxide per liter of alveolar volume (DLCO/VA) decrease were significantly higher in the IgG4-RRD group than in the IgG4-RDEI group (P=0.029). DLCO/VA were significantly higher in the IgG4-RDEI than in the IgG4-RRD group (P=0.044), but otherwise, there were no significant differences. We report the first finding of a negative correlation between pulmonary diffusing capacity and total serum concentrations of IgG and IgG subclasses (IgG4, IgG3 and IgG2). CONCLUSIONS: DLCO/VA plays an important role for detecting lung involvement in IgG4-RD patients. The patient with high serum IgG may be more prone to respiratory involvement.

Pulmonary function test findings in patients with acute inhalation injury caused by smoke bombs.

BACKGROUND: This study aimed to determine the effects of smoke bomb-induced acute inhalation injury on pulmonary function at different stages of lung injury. METHODS: We performed pulmonary function tests (PFTs) in 15 patients with acute inhalation injury from days 3 to 180 after smoke inhalation. We measured the trace element zinc in whole blood on days 4 and 17, and correlations of zinc levels with PFTs were performed. RESULTS: In the acute stage of lung injury (day 3), 3 of 11 patients with mild symptoms had normal pulmonary function and 8 patients with restrictive ventilatory dysfunction and reduced diffusing capacity. Some patients also had mild obstructive ventilatory dysfunction (5 patients) and a decline in small airway function (6 patients). For patients with severe symptoms, PFT results showed moderate to severe restrictive ventilatory dysfunction and reduced diffusing capacity. PaCO2 was significantly higher (P=0.047) in patients with reduced small airway function compared with those with normal small airway function. Whole blood zinc levels in the convalescence stage (day 17) were significantly lower than those in the acute stage (day 4). Zinc in the acute stage was negatively correlated with DLCO/VA on days 3, 10, and 46 (r=-0.633, -0.676, and -0.675 respectively, P<0.05). CONCLUSIONS: Smoke inhalation injury mainly causes restrictive ventilatory dysfunction and reduced diffusing capacity, and causes mild obstructive ventilatory dysfunction and small airway function decline in some patients. Zinc is negatively correlated with DLCO/VA. Zinc levels may be able to predict prognosis and indicate the degree of lung injury.

Estrogen plays a critical role in AAV2-mediated gene transfer in ovarian cancer.

AIM: The aim of our study was to develop an effective gene delivery system for ovarian cancer gene therapy. METHODS: The expression of heparin sulfate proteoglycan (HSPG) and integrins alpha(upsilon)beta(3) and alpha(upsilon)beta(5) were analyzed with flow cytometry on 2 human ovarian cancer cell lines (OVCAR-3 and SKOV-3ip). The gene transduction efficiencies were evaluated with recombinant adeno-associated viral vector (rAAV)2-green fluorescent protein or rAAV2-lactase Z followed by flow cytometry or cytohistochemistry staining. The effect of 17beta-estradiol on ovarian cancer cell proliferation, HSPG, the expressions of integrins alpha(upsilon)beta(3) and alpha(upsilon)beta(5), and adeno-associated viral vector (AAV)2-mediated gene transduction were determined. RESULTS: In the present study, we found: (1) a variation in HSPG and the expressions of integrins alpha(upsilon)beta(3) and alpha(upsilon)beta(5) between OVCAR-3 and SKOV-3ip; (2) that 17beta-estradiol was shown to significantly stimulate cell proliferation and integrin beta(5) expression in certain ovarian cancer cell lines; and (3) integrintargeted A520/N584RGD-rAAV2, which has alternative interactivity with integrins and abrogates the binding capacity HSPG, showed much higher gene transduction efficiency in ovarian cancer cells than rAAV2 in the presence/absence of 17beta-estradiol. Moreover, this RGD-modified rAAV2 exerted more efficient transduction in ovarian cancer cells in response to 17beta-estradiol. CONCLUSION: Our findings implied that A520/N584RGD-rAAV2 may offer great potential for ovarian cancer treatment in vivo.

Role and significance of pelvic four-contrast defecography in the diagnosis of outlet obstructive constipation / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study the changes in pelvic floor morphology and relationships of the pelvic organs in patients with outlet obstructive constipation (OOC).</p><p><b>METHODS</b>Fifty-nine OOC patients and 12 healthy volunteers were examined by simultaneous pelvic four-contrast defecography, including pelvicography, vaginal opacification, voiding cystography and defecography. The levels of perineum, peritoneum and bladder, and anorectal angle were measured and recorded.</p><p><b>RESULTS</b>Among the 59 OOC patients, 46 cases of internal rectal prolapse (IRP), 29 cases of pelvic floor hernia, 7 cases of rectocele (RC), 7 cases of spastic pelvic floor syndrome (SPFS), 5 cases of descending perineum syndrome were diagnosed by pelvic four-contrast defecography. Six cystoceles and 10 uterine prolapses were also found by pelvic four-contrast defecography. Compared with healthy volunteers, OOC patients had significantly large anorectal angles during defecation (P<0.05), abnormality descending of perineum during rest and defecation (P<0.05, P<0.01), and a deep pouch of Douglas during defecation (P<0.01). OOC patients combined with urinary system symptoms had an abnormal descent of the bladder during rest and defecation (P<0.05, P<0.01).</p><p><b>CONCLUSION</b>Pelvic four-contrast defecography is an effective method for the diagnosis of IRP, RC, pelvic floor hernia, peritoneocele, cystocele and uterine prolapse, and is helpful in the selection of proper surgical procedures for the treatment of OOC.</p>

Targeting gene-virotherapy for cancer.

Gene therapy and viral therapy for cancer have therapeutic effects, but there has been no significant breakthrough in these two forms of therapy. Therefore, a new strategy called "targeting gene-virotherapy", which combines the advantages of gene therapy and viral therapy, has been formulated. This new therapy has stronger antitumor effects than either gene therapy or viral therapy. A tumor-specific replicative adenovirus vector ZD55 (E1B55KD deleted Adv.) was constructed and various single therapeutic genes were inserted into ZD55 to form ZD55-gene. These are the targeting gene-virotherapy genes. But experiments showed that a single gene was not effective in eliminating the tumor mass, and therefore two genes were separately inserted into ZD55. This strategy is called "targeting dual gene-virotherapy" (with PCT patent). Better results were obtained with this strategy, and all the xenograft tumor masses were completely eliminated in all mice when two suitable genes producing a synergetic or compensative effect were chosen. Twenty-six papers on these strategies have been published by researchers in our laboratory. Furthermore, an adenoviral vector with two targeting promoters harboring two antitumor genes has been constructed for cancer therapy. Promising results have been obtained with this adenoviral vector and another patent has been applied for. This antitumor strategy can be used to kill tumor cells completely with minimum damage to normal cells.

[Relationship between cytotoxicity of two types of TNF-alpha and intracellular free calcium concentration in target cells].

AIM: To compare the changes of free calcium concentration in target cells killed by transmembrane tumor necrosis factor-alpha(TM-TNF-alpha) and secretory TNF-alpha(S-TNF-alpha). METHODS: The cytotoxicity of two types of TNF-alpha was tested by bioassay. The intracellular Ca(2+) concentration was determined by Frua-2. RESULTS: TM-TNF-alpha had cytotoxic effect on all 6 kinds of target cells, whereas S-TNF-alpha could kill only two of them. The cytotoxic activity of both types of TNF-alpha was accompanied by a dramatically increase of intracellular free Ca(2+) concentration. The intracellular Ca(2+) concentration in the target cells treated with S-TNF-alpha was obviously reduced by pretreating target cells with 10 micromol/L calcium chelator EGTA for 30 minutes (P<0.01) and the cytotoxicity of S-TNF-alpha was significantly inhibited (P<0.01), while the pretreatment with EGTA had no effect on the intracellular Ca(2+) concentration in the target cells treated with TM-TNF-alpha and the cytotoxicity of TM-TNF-alpha. CONCLUSION: These results suggest that both types of TNF-alpha increase Ca(2+) concentration in target cells by promoting the redistribution of intracellular free calcium and only S-TNF-alpha seems to be able to accelerate the influx of extracellular calcium into the target cells.

[Participation of ICAM-1 in the cytotoxicity of TM-TNF-alpha mediated by TNF receptor I].

AIM: To identify membrane-associated molecules involved in the tumoricidal cytotoxicity of TM-TNF-alpha and to explore the molecular mechanism underlying the tumoricidal cytotoxicity of TM-TNF-alpha and differences of biological effects between TM-TNF-alpha and sTNF. METHODS: Antibody blocking test and RT-PCR were used to evaluate the role of ICAM-1 and VCAM-1 in the cytotoxicity of TM-TNF-alpha mediated by TNFR I or TNFR II. RESULTS: After the ICAM-1 expressed on the membrane of MCF-7 cells(expressing TNFRI) was blocked, the cytotoxicity of TM-TNF-alpha to this cell lines decreased significantly, while blocking VCAM-1 had no effect. Blocking ICAM-1 or VCAM-1 expressed on the membrane of HL-60 cells(expressing TNFRII)had no significant effects on the cytotoxicity of TM-TNF-alpha to this cell lines. CONCLUSION: ICAM-1 participates in the cytotoxicity of TM-TNF mediated by TNFRI.