Research Progress on Microbial Community Succession in the Postmortem Interval Estimation.

The postmortem interval (PMI) estimation is a key and difficult point in the practice of forensic medicine, and forensic scientists at home and abroad have been searching for objective, quantifiable and accurate methods of PMI estimation. With the development and combination of high-throughput sequencing technology and artificial intelligence technology, the establishment of PMI model based on the succession of the microbial community on corpses has become a research focus in the field of forensic medicine. This paper reviews the technical methods, research applications and influencing factors of microbial community in PMI estimation explored by using high-throughput sequencing technology, to provide a reference for the related research on the use of microbial community to estimate PMI.

The pro-inflammatory and anti-inflammatory cytokine profile in peripheral blood of women with recurrent implantation failure.

Limited information is available on the balance state of pro- and anti-inflammatory cytokines in patients with recurrent implantation failure (RIF). This study assessed the pro- and anti-inflammatory cytokines in plasma of 34 patients with RIF, compared with those of 25 women with a successful pregnancy in the first IVF/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) cycle. The IFN-γ, IL-1ß, IL-6 and IL-4 concentrations were higher, whereas the TGF-ß1 concentration was lower in the RIF group compared with the control group. Furthermore, the ratios of pro-inflammatory and anti-inflammatory cytokines IFN-γ/IL-4, IFN-γ/IL-10, IFN-γ/TGF-ß1, IL-6/IL-10, IL-6/TGF-ß1, IL-1ß/TGF-ß1 and TNF-α/TGF-ß1 were higher in the RIF group (all P < 0.01). The results suggested a shift toward a pro-inflammatory state in peripheral blood of the patients with RIF.

Qualitative visual trichotomous assessment improves the value of fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography in predicting the prognosis of diffuse large B-cell lymphoma.

INTRODUCTION: Fluorine-18 fluorodeoxyglucose (18 F-FDG) positron emission tomography/computed tomography (PET/CT) is a powerful tool for monitoring the response of diffuse large B-cell lymphoma (DLBCL) to therapy, but the criteria to interpret PET/CT results remain under debate. We investigated the value of post-treatment PET/CT in predicting the prognosis of DLBCL patients when interpreted according to qualitative visual trichotomous assessment (QVTA) criteria compared with the Deauville criteria. METHODS: In this retrospective study, final PET/CT scans of DLBCL patients treated with rituximab-based regimens between October 2005 and November 2010 were interpreted using the Deauville and QVTA criteria. Survival curves were estimated using Kaplan-Meier analysis and compared using the log-rank test. RESULTS: A total of 253 patients were enrolled. The interpretation according to the Deauville criteria revealed that 181 patients had negative PET/CT scan results and 72 had positive results. The 3 year overall survival (OS) rate was significantly higher in patients with negative scan results than in those with positive results (91.6% vs. 57.5%, P<0.001). The 72 patients with positive scan results according to the Deauville criteria were divided into two groups by the interpretation according to the QVTA criteria: 29 had indeterminate results, and 43 had positive results. The 3 year OS rate was significantly higher in patients with indeterminate scan results than in those with positive results (91.2% vs. 33.5%, P<0.001) but was similar between patients with negative and indeterminate scan results (91.6% vs. 91.2%, P=0.921). CONCLUSIONS: Compared with the Deauville criteria, using the QVTA criteria for interpreting post-treatment PET/CT scans of DLBCL patients is likely to reduce the number of false positive results. The QVTA criteria are feasible for therapeutic outcome evaluation and can be used to guide risk-adapted therapy.

Cervical lymph node hyperplasia on [(18)F]-fluorodeoxyglucose positron emission tomography/computed tomography scan after treatment of children and adolescents with malignant lymphoma.

PURPOSE: To define imaging manifestations and clinical prognosis of cervical lymph node hyperplasia using [(18)F]-fluorodeoxyglucose positron emission tomography/computed tomography (FDG PET/CT) scanning after treatment of children and adolescents with malignant lymphoma. METHODS: Children and adolescent patients with malignant lymphoma who had high FDG uptake in their cervical lymph nodes via PET/CT after treatment, which was not due to tumor recurrence or residue, were retrospectively analyzed. RESULTS: Twenty-seven patients with a median age of 12 years were included; 11 had Hodgkin's disease and 16 had non-Hodgkin's lymphoma. The time from PET/CT scan to completion of therapy was 1-36 months, 85.2% (23/27) of which took place within 12 months. Three patients had confirmed lymph node follicular hyperplasia by biopsy, while all 27 patients achieved disease-free survival during the follow-up period. The maximum standardized uptake values (SUVmax) of cervical lymph nodes were 2.2-16.2 and the maximum short axis ranged from 0.3 to 1.2 cm. Cervical lymph node hyperplasia was noted in neck levels I-V, and neck level II bilaterally had the highest incidence (100%). Bilateral cervical lymph node hyperplasia was symmetrical in terms of both the SUVmax and affected locations. Thymic hyperplasia and nasopharyngeal lymphoid hyperplasia were both observed in 24 patients (88.9%). There was no relationship in terms of the SUVmax between cervical lymph nodes and thymic tissue, cervical nodes or nasopharyngeal lymphoid tissue. CONCLUSION: Cervical lymph node hyperplasia with high FDG uptake on PET/CT scans found after treating children and adolescents with malignant lymphoma can be benign processes. Awareness of this possibility may help avoid invasive procedures and over-treatment.

Complementary Roles of Squamous Cell Carcinoma Antigen and (18)F-FDG PET/CT in Suspected Recurrence of Cervical Squamous Cell Cancer.

PURPOSE: To assess the clinical value of FDG PET/CT and evaluate the complementary roles of serum squamous cell carcinoma antigen (SCCAg) and FDG PET/CT in the diagnosis of suspected recurrent of cervical squamous cell cancer. METHODS: Serum SCCAg levels were retrospectively reviewed in patients previously treated for cervical squamous cell carcinoma, who had suspected recurrence of cervical cancer and who had undergone FDG PET/CT scans. The clinical impact of elevated SCCAg (>1.5 ng/ml) and negative SCCAg (≤1.5 ng/ml) levels were analyzed based on the results of PET/CT and final diagnosis. RESULTS: The overall patient-based sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of PET/CT for the detection of tumor recurrence or malignancy were 100% (86/86), 80.8% (21/26), 95.5% (107/112), 94.5% (86/91) and 100% (21/21), respectively. Of the 112 patients included in this study, recurrence or malignancy was detected by PET/CT in 62 of the 64 patients with elevated SCCAg, compared to 24 of the 48 patients with negative SCCAg levels. The overall patient-based PPV, NPV, sensitivity and accuracy of SCCAg for the detection of tumor recurrence or malignancy were 96.9% (62/64), 50% (24/48), 72.1% (62/86) and 76.8% (86/112), respectively. The five false-positive PET/CT results were all associated with patients with negative SCCAg levels. The PPV of positive PET/CT-associated elevated SCCAg for the detection of tumor recurrence or malignancy was 100% (62/62). The NPV of negative SCCAg-associated negative PET/CT was 100% (19/19). CONCLUSIONS: Serum SCCAg evaluation and FDG PET/CT imaging can be complementary techniques in cases of suspected recurrent cervical squamous cancer. Positive PET/CT with elevated SCCAg can predict recurrence. Although PET/CT cannot confidently be deferred due to a negative SCCAg test, the possibility of a false-positive PET/CT in those cases may have diagnostic importance.

Increased circulating Th1/Th2 ratios but not other lymphocyte subsets during controlled ovarian stimulation are linked to subsequent implantation failure after transfer of in vitro fertilized embryos.

PROBLEM: To investigate alterations of circulating lymphocyte subsets in women undergoing controlled ovarian stimulation (COS) and survey their relations with pregnancy outcome. METHOD OF STUDY: Fifty-one women were included in this study, of which 24 women were successfully pregnant in the index IVF cycle (the successful pregnancy group, sPG) and 27 women failed to get pregnant (the non-pregnancy group, nPG). Peripheral blood was collected at three time points during COS: on the day before the first GnRHa administration (GnRHa day), on the day before the first Gn administration (Gn day), and on the day before HCG administration (HCG day). The intracellular Th1(IFN-γ and TNF-α)/Th2(IL-10) ratios in circulating Th cells after in vitro phorbol-12-myristate-13-acetate stimulation, the NK cytotoxicity, and the percentage of T, B and NK cell subsets were assessed by flow cytometer. A third group consisting of patients with recurrent implantation failure (RIF group, n = 18) was also enrolled, and the intracellular Th1/Th2 ratios were assessed at HCG day. RESULTS: Intergroup comparison: The ratios of IFN-γ/IL-10 and TNF-α/IL-10 at HCG day were significantly higher (P < 0.05) in nPG than those in sPG, while no change at GnRHa day or Gn day. Higher (P < 0.05) ratios of IFN-γ/IL-10 and TNF-α/IL-10 at HCG day were also observed in RIF group compared to those in sPG. No significant change (P > 0.05) was found in other assessed lymphocyte subsets or in the concentrations of progesterone, estradiol, luteinizing hormone, or follicle-stimulating hormone between the sPG and nPG at the three time points. Intragroup comparison: The ratios of IFN-γ/IL-10 and TNF-α/IL-10 at HCG day were significantly (P < 0.05) lower than those at GnRHa day in sPG, but not in nPG. No significant (P > 0.05) difference was found in other assessed parameters or at other time points. CONCLUSIONS: The ratios of Th1/Th2 were fluctuated during COS, and they might affect the pregnancy outcome of IVF. The circulating NK cytotoxicity and percentage of lymphocyte subsets were relatively stable during the COS.

Use of subsequent PET/CT in diffuse large B-cell lymphoma patients in complete remission following primary therapy.

Interim 18F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (I-PET/CT) is a powerful tool for monitoring the response to therapy in diffuse large B-cell lymphoma (DLBCL). This retrospective study aimed to determine when and how to use I-PET/CT in DLBCL. A total of 197 patients treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) were enrolled between October 2005 and July 2011; PET/CT was performed at the time of diagnosis (PET/CT0), after 2 and 4 cycles of chemotherapy (PET/CT2 and PET/CT4, respectively), and at the end of treatment (F-PET/CT). According to the International Harmonization Project for Response Criteria in Lymphoma, 110 patients had negative PET/CT2 scans, and 87 had positive PET/CT2 scans. The PET/CT2-negative patients had significantly higher 3-year progression-free survival rate (75.8% vs. 38.2%) and 3-year overall survival rate (93.5% vs. 55.6%) than PET/CT2-positive patients. All PET/CT2-negative patients remained negative at PET/CT4, but 3 were positive at F-PET/CT. Among the 87 PET/CT2-positive patients, 57 remained positive at F-PET/CT, and 32 progressed during chemotherapy (15 at PET/CT4 and 17 at F-PET/CT). Comparing PET/CT4 with PET/CT0, 7 patients exhibited progression, and 8 achieved partial remission. Comparing F-PET/CT with PET/CT0, 10 patients exhibited progression, and 7 achieved partial remission. In conclusion, our results indicate that I-PET/CT should be performed after 2 rather than 4 cycles of immunochemotherapy in DLBCL patients. There is a limited role for subsequent PET/CT in the detection of relapse in PET/CT2-negative patients, but repeat PET/CT is required if the PET/CT2 findings are positive.

[Application of 18F-FDG PET/CT in cervical cancer with elevated levels of serum squamous cell carcinoma antigen during the follow-up].

BACKGROUND AND OBJECTIVE: Accurate and early diagnosis of recurrence for cervical cancer after the treatment and aggressive salvage treatment could improve the prognosis of this disease. Serum squamous cell carcinoma antigen (SCCAg) is the most commonly used tumor marker for the detection of asymptomatic recurrence of cervical cancer. This study was to evaluate the application and value of (18)F-FDG PET/CT in cervical cancer with elevated of serum SCCAg level during the follow-up. METHODS: Thirty-one patients with cervical cancer with elevated serum SCCAg level during the follow-up undergoing (18)F-FDG PET/CT in Sun Yat-sen University Cancer Center between August 2005 and November 2008 were entered into this retrospective study. The pathological types, the serum SCCAg level, PET/CT results, results of other imaging modalities, pathological and clinical follow-ups were recorded. RESULTS: All 31 patients'pathological examination showed squamous cell carcinoma, including three adenosquamous carcinoma. Lesions of all patients were examined by PET/CT. Three patients had local recurrence in the uterus or vagina, 28 had metastatic disease. Of these 31 patients, three were confirmed to have local recurrent disease, 27 were verified to have metastatic disease and one was diagnosed as primary lung squamous cell carcinoma by pathological or clinical manifestations. The total detection rate of PET/CT for malignancy was 100% (31/31); the diagnostic accuracy of PET/CT for recurrent cervical cancer was 96.8% (30/31). The levels of serum SCCAg during the follow-up were 1.5-37.8 ng/ml. There was no relation between the level of serum SCCAg and the maximum standard uptake value (SUVmax) of PET/CT. Compared with other imaging modalities, PET/CT was more efficient in detecting recurrence and finding more lesions. CONCLUSIONS: An elevated level of SCCAg in cervical cancer during the follow-up indicates tumor recurrence. PET/CT is efficient in detecting the recurrence and has high diagnostic accuracy.

[18F-FDG PET/CT for the detection of primary tumors metastasizing to lymph nodes of the neck].

BACKGROUND AND OBJECTIVE: Lymph node metastases are commonly seen in malignant tumors of the head and neck. Detection of the primary tumors affects the quality of life and survival rates of these patients. This study was to evaluate the application of (18)F-FDG PET/CT in detecting primary tumors metastasizing to lymph nodes of the neck, and to assess the positive predictive value of (18)F-FDG PET/CT. METHODS: In total 93 patients with pathologically confirmed neck lymph node metastases from unknown primary tumors underwent (18)F-FDG PET/CT in Sun Yat-sen University Cancer Center between June 2005 to April 2008 were entered into this study. The primary tumors of patients were initially diagnosed according to different PET/CT standards as definite diagnosis, suspicious diagnosis and no signs of primary tumors. All diagnosis based on PET/CT images were verified by pathological exams or additional imaging tests. RESULTS: Forty cases made definite diagnosis by PET/CT were all confirmed by pathological or clinical exams, with a positive predictive value of 100%. Of 28 cases made suspicious diagnosis, 16 were pathologically confirmed, with a positive predictive value of 57.1%. Two patients, who were suggested lymph node metastases in mediastinum without the sign of the primary tumor by PET/CT, were clinically verified as primary mediastinal lung cancer. The primary tumors of another two patients were not detected by PET/CT, but were found under endoscopy. The total detection rate of PET/CT for the primary tumor was 60.2% (56/93). CONCLUSION: PET/CT is of important clinical value in detecting primary tumors metastasizing to lymph nodes of the neck.

[Determination of sperm DNA integrity by improved sperm chromatin structure assay test].

OBJECTIVE: To search for a simple, quick and accurate method for the sperm DNA integrity test by modifying the conventional sperm chromatin dispersion (SCD) test, and to investigate the reference value range of normal sperm DNA integrity by the improved SCD assay. METHODS: We modified the conventional SCD test in its procedures, reagents preparing and utensils to be used, compared the improved method with the conventional one, and detected 293 normal semen samples for sperm DNA integrity by the modified SCD test. RESULTS: An improved SCD test was established, and no statistical differences were found between the results of the modified and conventional methods. The reference value range was determined by the improved SCD assay for normal sperm DNA integrity, which was < 32.58% for abnormal sperm DNA integrity. CONCLUSION: The improved SCD test could detect sperm DNA integrity more quickly, and it reduced the cost in reagents.

[Role of 18F-FDG PET/CT in diagnosis and staging of nasopharyngeal carcinoma].

BACKGROUND & OBJECTIVE: Because (18)F-FDG PET/CT can display cancer lesion morphology, size and the relatinship of the cancer lesion to adjacent tissues, it is used in the diagnosis and classification of nasopharyngeal carcinoma (NPC). This study was to explore the application value of (18)F-FDG PET/CT in TNM staging by comparing the performance of MRI and PET/CT together with pathological results for some small lymph nodes. METHODS: Sixty-eight pathologically proven NPC patients were selected from Sep.2005 to Mar. 2007 in Sun Yat-sen University Cancer Center. All cases underwent both MRI and (18)F-FDG PET/CT examinations. PET/CT two-dimensional (2D) data were collected. Automatic dose tracking scanning and contrast-enhanced CT scanning were performed. (18)F-FDG was intravenously administrated at a dose of 3.7-5.5 MBq/kg. MRI T1W, T2W and T1W Gd-DTPA enhanced images were obtained. Neck lymph nodes of 10 NPC patients were pathologically examined. RESULTS: Nasopharyneal lesions of all 68 cases were all clearly displayed by both PET/CT and MRI. Among 138 small lymph nodes smaller than 1cm in diameter, which were positively detected by PET/CT, only 28.0% were suggested questionable by MRI scanning. Fourteen out of sixteen (87.5%) positive lymph nodes detected by PET/CT from 10 patients were pathologically confirmed. Both PET/CT and MRI revealed enlarged lymph nodes of the neck. Some lymph nodes whose proliferation was inhibited during radiotherapy suggested by PET/CT were displayed intensified images by contrast-enhanced PET/CT and MRI. Metastases to lung, bone and liver in eight stage IVb patients were clearly displayed by PET/CT, but not by MRI. The stage of 24 patients was adjusted after PET/CT scanning. CONCLUSIONS: (18)F-FDG PET/CT with automatic dose tracking scanning protocol and contrast-enhanced scanning can provide more comprehensive information than MRI in diagnosing and staging NPC.

[The effect of p38 on the cycloheximide-induced HL-60 cell death through mitochondria pathway].

OBJECTIVE: To study the effect of p38 on the cycloheximide (CHX)-induced HL-60 cell death through mitochondria pathway. METHODS: Inhibition of p38 pathway was by SB203580 (SB). Four groups were set up: control, SB only, CHX only and SB + CHX. Sub-diploid cell ratio was detected by PI staining flow cytometry at 6, 9, 12, 18, 24 h time points, and apoptotic cell ratio by Annexin V-FITC/PI double staining flow cytometry at 6 h and 18 h time points. High J-aggregate cells were evaluated by the J-aggregate contents, measurement of the J-aggregate (FL2) and J-monomer (FL1) by JC-1 flow cytometry, calculation of the delta psi m by FL2/FL1 and analysis of the delta psi m changes at 18 h time points. RESULTS: The sub-diploid cell ratio in CHX group was significantly higher than that in control group at 6 h time point, and the ratio in SB + CHX group was significantly higher than that in CHX group at 9 h time point. At 18 h time point the apoptotic cell ratios in both CHX and SB + CHX groups were significantly higher than those in control group (P < 0.01). There was no significant difference of apoptotic cell ratio between CHX group and SB + CHX group (P > 0.05). At 18 h time point the necrotic cell ratios in both CHX and SB + CHX groups were significantly higher than that in control group (P < 0.01); and that in SB + CHX group was significantly higher than that in CHX group (P < 0.01). The high J-aggregate cell ratios in CHX and SB + CHX groups were significantly lower than that in control group (P < 0.05), and that was signficantly lower in SB + CHX group than in CHX group (P < 0.01). For the FL2/FL1 value (delta psi m) CHX group (0.17 +/- 0.01) and SB + CHX group (0.05 +/- 0.003) were significantly higher than control group (0.38 +/- 0.02) (P < 0.01), and SB + CHX group was significantly lower than CHX group (P < 0.01). CONCLUSION: CHX can induce HL-60 cell apoptosis and the cell mitochondria depolarization, and the latter was intensified by inhibition of the p38 pathway. p38 pathway may related to the cell necrosis in the cycloheximide-induced HL-60 cell apoptosis model. s

[Study on the mitochondrial inner membrane potential of apoptotic thymocytes at cell and organelle level].

In this study, dexamethasone was used to induce mouse thymocyte apoptosis. The PI and Annexin V/PI staining flowcytometry methods were adopted to detect late and early phases apoptosis. Inner mitochondrial membrane potential (deltapsim) was examined by JC-1 and DiOC6(3)/PI staining flowcytometry. Direct JC-1 staining technology was applied to test the of deltapsim abstracted pure mitochondria. Results showed: DEX could significantly induce late and early phase mouse thymocyte apoptosis; at cell level, DEX was observed to reduce staining ability of deltapsim dependant fluorescence, J-aggregate and DiOC6(3), to drop down mitochondria number, but to cause no significant change of cell membrane integrity. Results of pure mitochondria detection showed most of them maintained normal deltapsim. According to above results, we concluded DEX could reduce mitochondria number when inducing mouse thymocyte apoptosis; and the remaining ones maintain normal function to meet the energy need for apoptotic process.

[Change of mitochondria during apoptosis of HL-60 cells induced by camptothecine].

AIM: To study the changes of mitochondrial mass and membrane potential (deltapsi(m)) during apoptosis of human promyelocytic leukemia cells (HL-60) induced by camptothecine(CPT). METHODS: HL-60 cells were stimulated with 4x10(-6) mol/L CPT. Apoptosis and necrosis of HL-60 cells were detected by annexin V-FITC/PI staining. Mitochondrial mass and membrane potential (deltapsi(m)) were measured by NAO and DiOC(6)(3) stanings, respectively. RESULTS: After 12 hour treatment with 4x10(-6) mol/L CPT, the early apoptotic rate of HL-60 cells was significantly increased compared with the control group (12.75+/-4.61)% vs (2.88+/-2.49)%,(P<0.01), and the necrotic rate was significantly elevated too, (3.48+/-1.67)% and (0.71+/-1.10)%, (P<0.01). The result of PI staining showed that the apoptotic rate of HL-60 cells at late phase (12 h) in CPT group was (3.52+/-1.07)%, whereas that in control group was (0.46+/-1.06)%. At the same time, we observed that the cells in G(2)/M phase were arrested. The percentage of cells in G(2)/M phase in CPT group and the control was (13.45+/-1.91)% and (22.46+/-2.19)% (P<0.01) respectively. The percentage of cells with low mitochondrial mass in CPT and control groups was (25.74+/-2.09)% and (4.53+/-1.26)%, respectively (P<0.01). Mitochondrial membrane potential in CPT group and control group was (17.71+/-5.23)% and (1.64+/-2.00)%, respectively. CONCLUSION: During CPT-induced apoptosis of HL-60 cells, mitochondrial mass and membrane potential drop significantly, but its depolarization heightens.

[Postoperative adjuvant therapy for 189 patients with completely resected non-small cell lung cancer of stage III(A)].

BACKGROUND & OBJECTIVE: Postoperative adjuvant therapy for patients with completely resected non-small cell lung cancer (NSCLC) of stage III(A) is a hot research topic. This study was to discuss the influence of postoperative adjuvant radiotherapy and chemotherapy on survival time of patients with completely resected NSCLC of stage III(A). METHODS: Clinical data of 189 patients with completely resected NSCLC of stage III(A) treated in our hospital from Jan. 1995 to Jan. 2000 were retrospectively analyzed to evaluate the influence of postoperative radiotherapy and chemotherapy on patients' survival time. RESULTS: Median survival time of 68 patients with squamous cell carcinoma was significantly higher than that of 121 patients with adenocarcinoma (43.8 months vs. 26.8 months, P=0.038). Of 68 patients with squamous cell carcinoma, median survival time of 21 patients treated by surgeon alone had no significant difference with that of 28 patients received postoperative radiotherapy (42.8 months vs. 45.5 months, P=0.068), and 19 patients received postoperative chemotherapy (42.8 months vs. 52.0 months, P=0.075). Of 121 patients with adenocarcinoma, median survival time of 41 patients treated by surgeon alone has no significant difference with that of 43 patients received postoperative radiotherapy (26.7 months vs. 26.6 months, P=0.555), and 37 patients received postoperative chemotherapy (26.7 months vs.37.9 months, P=0.242). CONCLUSIONS: Pathologic type is the main factor that may affect survival time of patients with resectable NSCLC of stage III(A). Although no statistical significance appeared, postoperative adjuvant chemotherapy tend to prolong patients' survival time.