The viable but noncuturable (VBNC) state is a survival strategy when bacteria are exposed to en- vironment stress. The VBNC state is part of the life cycle of non-differentiating bacteria, and it has a far-reaching impact on traditional bacteriology. Cells in the VBNC state fail to grow on the routine bacterio- logical media, here its significance in human health and environment science are detailed. Cells entering the VBNC state exhibit dwarfing and a number of metabolic changes in respiration rates and macromolecular synthesis. This paper summarized the variations in DNA and protein comparing to the culturable cells. We also discussed the ability of cells to resuscitate from the VBNC state and return to an actively metabolizing and culturable form. Some new methods for monitoring the VBNC state were listed. Finally the future was suggested.

To standardize the harvest ways of Dendrobium officinale and improve the quality and yield of D. officinale, a field experiment was carried out to study the effect of two kinds of harvest ways, which were keeping some of the axial shoot and harvesting all of the shoot by the end of the year. Then, the agronomic traits and yield were measured and the contents of polysaccharides and extractum were determined. The results showed that the harvest ways significantly affected the growth of D. officinale. Keeping some of the axial shoot could significantly improved the number of sprout, stem length, internode number and the internodal length, which also triggered increase the weight of fresh stems, leaves and the total of them and dry stems in per unit area, but it could not promote the stem diameter and the polysaccharide content in stems. Keeping some of the axial shoot moderately was conducive to the improvement of the production of medicinal materials in the process of harvesting by promoting the germination and growth of new buds, and to ensure the polysaccharide content by regulating the illumination and the density of cultivation.
Agriculture ; methods ; Dendrobium ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development

Objective:The recombinant human retinal pigment epithelium-derived factor(PEDF)protein to be obtained and the angiogenesis of the rPEDF to be identified.Methods: PEDF gene gene was amplified by PCR and cloned into pET32a,rPEDF protein was expressed in E.coli BL21 and confirmed by SDS-PAGE and Western blot.The rPEDF was purified by Ni-NTA on denature condition.The concentration of the rPEDF was determined by Bradford method.The angiogenesis of the rPEDF was determined by chick chorioallantoic membrane(CAM) method.Results: The expression plasmid pET32a-PEDF was constructed successfully.The rPEDF was expressed with stable efficiency in E.coli BL21.The results of the CAM experiment showed that the rPEDF had notable angiogenesis effect in the concentration 0.4、0.04 ng/ml,but had no effect in 4 ng/ml.Conclusion:The PEDF gene was cloned and expressed efficiently,the angiogenesis of the rPEDF to be identified and the activity was worked in certain range.The results can facilitate studying its function and spreading its application.

OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

Men with non-obstructive azoospermia (NOA) can achieve fertility by testicular sperm extraction (TESE) coupled with intracytoplasmic sperm injection (ICSI), the key to which is the successful retrieval of sperm from the testis. Although improved testicular sperm extraction techniques have increased the chances of sperm retrieval, to predict preoperatively the success of sperm retrieval from NOA patients remains challenging. A non-invasive diagnostic technique predicting the presence of sperm in the testis would be useful for avoiding possible surgical intervention. At present, some preoperative variables, such as serum FSH, inhibin B level, testis volume, genetic analysis, histopathology on diagnostic biopsy, Raman Spectroscopy, and molecular and protein markers, have provided new insights into the chances of successful sperm retrieval in NOA males. This review aims to evaluate the preoperative factors currently available for predicting the outcomes of sperm retrieval from NOA patients.
Azoospermia ; therapy ; Biomarkers ; Biopsy ; Genetic Testing ; Humans ; Inhibins ; blood ; Male ; Sperm Injections, Intracytoplasmic ; Sperm Retrieval ; Spermatozoa ; cytology ; Testis ; cytology

OBJECTIVE: To eludicate the risk factors of mechanical ventilation and prolonged mechanical ventilation in patients with severe multiple injuries. METHODS: Consecutive patients with severe multiple injures who were treated in Peking University People's Hospital Trauma Medical Center between December 2016 and December 2019 were enrolled in this restropective chart-review study. According to mechanical ventilation and ventilatory time, the patients were divided into mechanical ventilation (MV) group and non-mechanical ventilation (NMV) groups, prolonged mechanical ventilation (PMV) group and shortened mechanical ventilation (SMV) groups. Clinical data such as gender, age, base excess, mechanism of injury, Glasgow Coma Scale (GCS), abbreviated injury scale (AIS) and injury severity score (ISS) were collected. To indentify the risk factors of mechanical ventilation and prolonged mecha-nical ventilation, univariate and multivariate Logistic analyses were carried out. RESULTS: In the present study, 112 patients (82 male, 30 female) with severe multiple injuries having a median age of 52 (range: 16-89 years) and a median ISS of 34 (range: 16-66) were enrolled. The primary mechanism of injury was traffic accident injury and falling injury. In the study, 62 and 50 patients were assigned to MV and NMV groups, respectively. Logistic analysis showed that GCS (OR=0.72, 95%CI: 0.53-0.92, P=0.03), base excess (OR=0.56, 95%CI: 0.37-0.88, P=0.002) and multiple rib fracture (OR=1.72, 95%CI: 1.60-2.80, P=0.012) were independent significant risk factors for mechanical ventilation after severe multiple injuries. Within the mechanical ventilation group, 38 and 24 patients were assigned to PMV and SMVgroups, respectively. Compared with the SMV group, the PMV group had a higher ISS and higher rate of severe head trauma. The length of hospital stay of PMV group was longer than that of SMV groups. Meanwhile, the incidence of tracheotomy in PMV group was high. CONCLUSIONS GCS, base excess and rib fracture might be independent risk factors for mechanical ventilation. Higher ISS and lower GCS might prolong the ventilatory time and the length of hospital stay. Meanwhile, the incidence of tracheotomy was high in PMV group because of the longer ventilatory time and poor consciousness.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Injury Severity Score ; Male ; Middle Aged ; Multiple Trauma ; Respiration, Artificial ; Retrospective Studies ; Risk Factors ; Young Adult

OBJECTIVETo investigate the clinicopathological features of primary gastrointestinal non-Hodgkin's lymphomas (PGI-NHL) and their prognostic values.

METHODSThe clinical and pathological data of 216 patients diagnosed as PGI-NHL from Zhejiang Cancer Hospital were analyzed retrospectively. χ² test, log-liner model analysis, COX proportional hazard regression analysis and Life-table survival analysis were used to analyze the survival status of the patients by SAS 8.2 software, and Log-rank test was performed to couple the overall survival rates with different prognostic factors.

RESULTSTotally, the age of onset was 8 to 89 years with the median age of 56.5 years. Male versus female was 1.27∶1(121∶95). The most frequently involved location was stomach (147 cases, 68.1%), followed by ileocecus (25 cases, 11.6%), large intestine (20 cases, 9.3%), small intestine (17 cases, 7.9%) and multiple GI involvement (5 cases, 2.3%). 182 cases were classified as B cell lymphomas, 22 cases as T cell lymphomas, and 12 cases not classified exactly due to insufficient data. The 3-year and 5-year survival rates of the patients were 69.4% and 53.3%, respectively. Univariate analysis revealed that age＞60 years, ECOG≥2, high LDH level, stage Ⅲ-Ⅳ, IPI≥2, T cell type and intestinal involvement were predictors for poor prognosis. IPI≥2, T cell type and intestinal involvement were independent adverse predictors for prognosis by multiple COX proportional hazard regression analysis. Among different treatment groups, cases received chemotherapy combined with local radiotherapy gained the best survival status.

CONCLUSIONB-cell lymphoma was the main pathological type in PGI-NHL; IPI≥2, T-cell type and intestinal involvement are independent adverse predictors for prognosis; chemotherapy combined with local radiotherapy might be the choice of approach for advanced stage and aggressive PGI-HNL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Gastrointestinal Neoplasms ; drug therapy ; pathology ; Humans ; Lymphoma, Non-Hodgkin ; drug therapy ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult

Purpose To study the clinical features, immunophenotypes and prognostic factors of primary breast diffuse large B-celllymphoma (PBDLBCL). Methods The clinical pathological data of 49 cases of PBDLBCL during January 2006 to December 2016 were retrospectively analysed, and the basic clinical and pathologic data, pathologic types and the immunohistochemical slides by EnVision method for staining were summarized. Results 47 cases were women and 2 cases were men. The age ranged from 24 to 79 year old with the median age of 48 year old. On microscopic observation, tumor cells were large to medium-sized which characterized as diffuse infiltration between the lobules of mammary gland, around the duct, interstitial and fat tissue, some were single file cord pattern. The immunophenotype showed 37 cases were of non-GCB, 12 cases were GCB type. Ki-67 index were greater than 40％. According to Ann Arbor staging, 16 cases were stage I EA, 28 cases were stage Ⅱ EA, 5 cases were stage Ⅳ E. IPI score: 30 cases with 0 ～1 score, 10 cases with 2 score, 9 cases with 3 score. Patients were followed up from 5 to 146 months, The 3-year overall survival (OS) rate was 51.2％ and 5-year OS rate was 36.7％, Single factor analysis showed that there were statistically significant difference in clinical stage, levels of LDH, IPI score, BCL-2 protein expression, and BCL-6 protein expression in 3 and 5 years of OS rate. The multiple factor analysis of Cox regression showed that the increase of IPI risk classification was the independent adverse prognostic factor of primary breast diffuse large B-cell lymphoma. Conclusion The diagnosis of PBDLBCL is confirmed by pathological biopsy and immunohistochemical markers. The immunophenotype was mainly non-GCB type. Comprehensive treatment including surgery, chemotherapy and radiotherapy is appropriate. The prognosis should be comprehensively evaluated by multiple factors. IPI increase risk classification is the independent adverse prognostic factor.

OBJECTIVETo explore the value of dual-time-point (18)F-fluorodeoxyglucose integrated positron emission and computed tomography ((18)F-FDG PET-CT) in differentiation of malignant from benign gastrointestinal diseases.

METHODSSixty five patients with suspected gastrointestinal lesions underwent dual-time-point (18)F-FDG PET-CT imaging. Standardized uptake value (SUV) was calculated for semi-quantitative assessment. The SUV of the two acquisitions were signed SUV(early) and SUV(delayed), respectively. Then the change of SUVmax (ΔSUVmax) was calculated. The ROC curves of the SUV(early), SUV(delayed) and ΔSUV were drawn to find the best cut-off point value for differential diagnosis, and then the sensitivity, specificity, positive predictive value, negative predictive value and accuracy were calculated, respectively.

RESULTSOf the malignant lesions, the SUVmax in delayed imaging were significantly higher than those in early imaging, while there were no significant differences of SUVmax between the two images of the benign lesions. The ΔSUVmax of the malignant lesions were significantly higher than that of the benign ones. Taking the SUVmax higher than 9.2 in early imaging as positive diagnostic criteria, the sensitivity was 72.7%, the specificity was 85.7%, the positive predictive value was 91.4%, the negative predictive value was 60.0%, and the accuracy was 76.9%. Taking the SUVmax higher than 10.9 in delayed imaging as positive diagnostic criteria, the sensitivity was 75.0%, the specificity was 90.5%, the positive predictive value was 94.3%, the negative predictive value was 63.3%, and the accuracy was 80.0%. Taking the ΔSUVmax higher than 5.1% as positive diagnostic criteria, the sensitivity was 95.5%, the specificity was 85.7%, the positive predictive value was 93.3%, the negative predictive value was 90.0%, and the accuracy was 92.3%. The accuracy of dual-time-point (18)F-FDG PET-CT imaging was significantly higher than that of single-time point (18)F-FDG PET-CT imaging.

CONCLUSIONDual-time-point (18)F-FDG PET-CT imaging is a useful method for differentiating malignant from benign gastrointestinal diseases, and it is superior to the single-time point (18)F-FDG PET-CT imaging.

Adenocarcinoma ; diagnosis ; pathology ; Adult ; Aged ; Aged, 80 and over ; Colitis ; diagnosis ; pathology ; Colorectal Neoplasms ; diagnosis ; pathology ; Diagnosis, Differential ; Female ; Fluorodeoxyglucose F18 ; Follow-Up Studies ; Gastritis ; diagnosis ; pathology ; Gastrointestinal Diseases ; diagnosis ; pathology ; Gastrointestinal Neoplasms ; diagnosis ; pathology ; Humans ; Male ; Middle Aged ; Positron-Emission Tomography ; methods ; Predictive Value of Tests ; Proctitis ; diagnosis ; pathology ; Proctocolitis ; diagnosis ; pathology ; ROC Curve ; Radiopharmaceuticals ; Sensitivity and Specificity ; Stomach Neoplasms ; diagnosis ; pathology ; Tomography, X-Ray Computed ; methods

OBJECTIVETo investigate the usefulness of (18)F-FDG PET/CT imaging in restaging, evaluating the treatment outcome, monitoring relapse and predicting prognosis of T-cell lymphoma.

METHODSRetrospective analysis of PET/CT image results of thirty-four patients with T-cell lymphoma, and to evaluate its clinical significance in restaging, treatment efficiency, relapse monitor and prognosis prediction.

RESULTSClinical restaging among the 20 stage I and II patients, 6 were ascended, 9 descended and 5 unchanged. Restaging among the other 14 stage III and IV patients, 3 were ascended, 4 descended and 7 unchanged. There were 12 patients in complete remission (CR), 11 in partial remission (PR), 2 in stable disease (SD) and 9 in progressive disease (PD) among all the 34 patients. There is obvious statistical difference of the standardized uptake value (SUV) between the efficacy group and the inefficacy group after treatment of 6 courses at least in 25 patients among all the 34 patients (P = 0.009). There is obvious statistical difference of the SUV value before and after treatment in 8 patients among all the 34 patients (P = 0.000). There is obvious statistical difference in the survival time between the efficacy group and the inefficacy group after treatment of 6 courses at least in 25 patients among all the 34 patients (P = 0.015).

CONCLUSIONS(18)F-FDG PET/CT imaging plays an very important role in guiding clinical restaging, evaluating the treatment outcome, monitoring relapse and predicting prognosis of T-cell lymphoma. It is helpful to establish personalized treatment planning.

Adolescent ; Adult ; Aged ; Female ; Fluorodeoxyglucose F18 ; Humans ; Lymphoma, T-Cell ; diagnostic imaging ; therapy ; Male ; Middle Aged ; Positron-Emission Tomography ; methods ; Prognosis ; Retrospective Studies ; Tomography, X-Ray Computed ; methods ; Treatment Outcome ; Young Adult