Application of Automatic Nucleic Acid Extractor Combined with Vacuum Concentrator in Forensic Science / 法医学杂志

Objective To explore the application value of automatic nucleic acid extractor combined with vacuum concentrator in forensic DNA extraction. Methods Gradient samples of human peripheral venous blood were collected at 40, 80, 120, 160, 200, 240, 280 and 320 fold dilution. The samples of each gradient were treated with no inhibitor, black oil, rust, fruit acid, tin foil and indigo, respectively. The automatic nucleic acid extractor was used for DNA purification and extraction of the above samples. The extracted DNA eluent （6 μL） was taken for amplification directly, and the rest was concentrated by vacuum concentrator. DNA was amplified and examined using the Investigator 26plex QS kit before and after concentration. Results Only gradient samples treated with fruit acid obtained complete STR typing results at 40 fold dilution. The other 5 methods obtained complete STR typing results at 40-160 fold dilution. The results of STR typing after DNA concentration showed that the average peak height and detection rates of gene loci both increased to a certain extent, but the effect was not obvious. Conclusion The automatic nucleic acid extractor has an efficient inhibitor removal ability and high extracting efficiency of DNA. The vacuum concentrator can concentrate DNA samples to a certain extent. Combining the automatic nucleic acid extractor with the vacuum concentrator can improve the examination success rate of forensic materials.

Effect of Shaoyaotang on Expressions of TLR4, NF-κB p65 and IL-6 in Rats with Damp-heat Ulcerative Colitis / 中国实验方剂学杂志

Objective:To explore the mechanism of Shaoyaotang in the treatment of ulcerative colitis (UC) based on toll-like receptor 4 (TLR4)/nuclear factor kappaB (NF-κB) signaling pathway. Method:A total of 50 Wistar rats were selected, including half male and half female. The damp-heat UC rat model was replicated by the methods of the combination of diseases and syndromes and the combination of 2, 4, 6-nitrobenzene sulfonic acid (TNBS) and ethanol. After the successful modeling, the model rats were randomly divided into model group, salazulesulfonate group, and low, medium and high-dose Shaoyaotang groups, and 10 rats (half male and half female) were selected as the blank control group. Low, medium and high-dose Shaoyaotang groups were given 6, 12, 24 g·kg-1 by gavage, and salazonyl arsenic group was given 1 g·kg-1 by gavage. Blank control group was given the equal volume of normal saline for 21 consecutive days. Colon samples were collected after the last administration, and the expressions of TLR4, NF-κB p65 and IL-6 mRNA in colon tissues were detected by fluorescent quantitative polymerase chain reaction (Real-time PCR）, and the expressions of TLR4, NF-κB p65 and IL-6 protein in colon tissues were detected by Western blot. Result:Compared with the blank control group, the relative expressions of TLR4, NF-κB p65, IL-6 mRNA and protein in the model group were significantly increased (P<0.05). Compared with the model group, the expression levels of TLR4, NF-κB p65 and IL-6 mRNA and protein in the salazopyridine group and Shaoyaotang groups were significantly decreased (P<0.05). Conclusion:Shaoyaotang can inhibit the development of UC by regulating the expressions of TLR4, NF-κB p65 and IL-6 mRNA and proteins in the TLR4/NF-κB pathway.

Quantitative analysis of residual ethanol content in corneal epithelium flaps in LASEK / 眼科新进展

Objective To detect and analyze of residual ethanol in abandoned flaps after laser subepithelial keratomileusis (LASEK) with ethanol infiltration methods.Methods Together 20 patients (40 eyes) undergoing LASEK were recruited in the study.After infiltrated with 20％ ethanol and rinsed in equilibration solution,the corneal epithelial free flap was isolated and removed in time for sealing,and then procedures were continuously completed.Finally,observation of the skin flap production,postoperative irritation symptoms,epithelial healing,visual recovery and postoperative haze situation was performed,and then the amount of ethanol in the epithelial flap was measured.Results There was no failure in making the intact corneal flaps.The sensory score of postoperative irritation was 2.52 ± 1.46.Neonatal epithelial with 1 grade was observed in 32 eyes,2 grade in 8 eyes 5 days after surgery,while corneal haze with 0.5 grade was occurred in 3 eyes,1 grade in 2 eyes 12 weeks after surgery.There were ethanol residues in corneal epithelium in the abandoned flaps,with the amount of ethanol residues of (0.205 2 ± 0.041 0) μL in each flap.Conclusion It is found that a certain amount of ethanol residue in the corneal epithelium after LASEK with ethanol infiltration equilibration solution rinse,which may be one reason of the corneal irritation symptoms and corneal haze.

China Patient-centered Evaluative Assessment of Cardiac Events Prospective Study of Acute Myocardial Infarction: Study Design / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Despite the rapid growth in the incidence of acute myocardial infarction (AMI) in China, there is limited information about patients' experiences after AMI hospitalization, especially on long-term adverse events and patient-reported outcomes (PROs).</p><p><b>METHODS</b>The China Patient-centered Evaluative Assessment of Cardiac Events (PEACE)-Prospective AMI Study will enroll 4000 consecutive AMI patients from 53 diverse hospitals across China and follow them longitudinally for 12 months to document their treatment, recovery, and outcomes. Details of patients' medical history, treatment, and in-hospital outcomes are abstracted from medical charts. Comprehensive baseline interviews are being conducted to characterize patient demographics, risk factors, presentation, and healthcare utilization. As part of these interviews, validated instruments are administered to measure PROs, including quality of life, symptoms, mood, cognition, and sexual activity. Follow-up interviews, measuring PROs, medication adherence, risk factor control, and collecting hospitalization events are conducted at 1, 6, and 12 months after discharge. Supporting documents for potential outcomes are collected for adjudication by clinicians at the National Coordinating Center. Blood and urine samples are also obtained at baseline, 1- and 12-month follow-up. In addition, we are conducting a survey of participating hospitals to characterize their organizational characteristics.</p><p><b>CONCLUSION</b>The China PEACE-Prospective AMI study will be uniquely positioned to generate new information regarding patient's experiences and outcomes after AMI in China and serve as a foundation for quality improvement activities.</p>

Glassy cell carcinoma of cervix: a clinicopathologic analysis of 5 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathological characteristics, histological diagnosis, immunohistochemistry and prognosis of cervical glassy cell carcinoma (GCC).</p><p><b>METHODS</b>The clinical characteristics, cytology, histology and immunohistochemistry were analyzed in 5 cases of GCC.</p><p><b>RESULTS</b>The average age of the five patients was 34.4 years (31 - 41 years). Abnormal vaginal bleeding and/or watery discharge were clinical presentations. One case was complicated with pregnancy and another one had a seven-year history of using contraceptives. All patients had an obvious mass in the cervix. Characteristic morphological features of GCC were present in 2 cases. Morphologically, the tumors consisted of clusters of tumor cells with distinct cell bounders, a large amount of eosinophilic granules in the cytoplasm imparting ground glass appearance, and thin nuclear membrane and prominent nucleoli. Nuclear enlargement and multinucleation were frequently noted. Mitosis and apoptosis were common. Numerous eosinophils and plasma cells were present in the stroma. Immunohistochemically, GCC expressed markers for both squamous cell carcinoma (p63 and CK34βE12) and adenocarcinoma (CAM5.2, MUC1, MUC2 and CEA). Ki-67 proliferation index was high (≥ 70%). All the five patients were treated with radical hysterectomy, followed by radiation and chemotherapy. The tumor-free survival time ranged from 25 days to 33 months.</p><p><b>CONCLUSIONS</b>GCC is a distinct variant of adenosquamous carcinoma of the cervix with high proliferation index and expression of markers of both squamous cell carcinoma and adenocarcinoma. The tumor has characteristic cytological and histological features.</p>

Expression of survivin, caspase-3 in oral precancerous lesions and oral squamous-cell carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the roles of surviving and caspase-3 in the development of oral cancer.</p><p><b>METHODS</b>Archival tissue sections of 17 oral squamous cell carcinoma (OSCC), 28 oral leukoplakia with dysplasia, 10 normal oral mucosa were obtained from Capital Medical University School of Stomatology for immunohistochemical staining of markers of survivin and caspase-3. The cell apoptosis was detected with terminal deoxynucleotidyl transferase-mediated nucleotide shift enzyme (TdT) mediated d-UTP end labeling (TUNEL). Positively stained cells were counted and analyzed statistically to determine potential relationship between survivin, caspase-3 and cell apoptosis.</p><p><b>RESULTS</b>The expression of survivin was faint or negative in normal epithelial cells. The average positive rate of survivin was (1.05 ± 1.21)% in control group and (21.89 ± 10.45)% in OSCC. Caspase-3 was expressed in all the normal mucosa,but it obviously down-regulated in dysplasia and OSCC. The apoptosis index (AI) decreased from (0.89 ± 0.46)% in normal mucosa to (0.21 ± 0.12)% in OSCC.</p><p><b>CONCLUSIONS</b>Both survivin and caspase-3 are associated with carcinogenesis of the oral mucosa. Survivin may restrain cell apoptosis by inhibiting caspase-3.</p>

Prediction of mycophenolic acid exposure in renal transplantation recipients by artificial neural network / 药学学报

The paper is aimed to establish an artificial neural network (ANN) for predicting mycophenolic acid (MPA) area under the plasma concentration-time curve (AUC) in renal transplantation recipients. 64 Chinese renal transplantation recipients receiving mycophenolate mofetil (MMF) were investigated. 10 timed samples were drawn at different days after transplantation. Plasma MPA concentration was determined by HPLC method and area under curve over the period of 0 to 12 h (AUC(0-12 h)) was calculated using the linear trapezoidal rule. ANN was established after network parameters were optimized using momentum method in combination with genetic algorithm. Furthermore, the predictive performance of ANN was compared with that of multiple linear regression (MLR). When using plasma MPA concentration of 0, 0.5, 2 h after MMF administration to predict MPA AUC(0-12 h), mean prediction error and mean absolute prediction error were -1.53% and 9.12%, respectively. Accuracy and precision of prediction by ANN were superior to that of MLR prediction, and similar results could be found when using plasma MPA concentration of 0, 0.5 h to predict MPA AUC(0-12h). The accuracy and precision of ANN prediction were superior to that of MLR prediction, and ANN can be used to predict MPA AUC(0-12 h).

Amifostine used in the treatment of patients with myelodysplastic syndrome / 中国实验血液学杂志

The study was aimed to investigate the curative effects and adverse effects of amifostine in the treatment of patients with myelodysplastic syndrome (MDS). Amifostine (AMF) was used alone (4/12) or combined with recombinant human erythropoietin (rh-EPO) (8/12) in 12 MDS patients. The therapeutic regimen was adopted with AMF 0.4 g/day for 5 days, then took a break of 2 days and then went on for 3 weeks consecutively, that was reputed as one treatment cycle. rh-EPO 6 000 U was used for 3 days per week. The results showed that 12 patients all attained hematological improvement in peripheral blood. 11 cases showed major effective response rate (91.7%), while 1 case showed minor response rate (8.3%). The effective response rate of hemoglobin, leukocytes and platelets was 100%, 75% and 58.3% respectively. The intervals of red cell transfusions (RCT) in 2 cases living on red cell transfusion before AMF treatment were prolonged after AMF treatments, and the amount of each RCT was decreased obviously. The side effect was usually discomfort of digestive system, but all patients can endure. In conclusion, Amifostine is a potential drug in the treatment of MDS patients with safety especially to those elder patients who often suffered from other multiple organ disfunctions, and the curative effect will be improved by more treatment cycles.

Immunohistochemical study of benign lymphoadenosis of oral mucosa / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the expression of Ki-67 and the changes of MVD and apoptosis in benign lymphoadenosis of oral mucosa (BLOM).</p><p><b>METHODS</b>The expression of Ki-67, CD34 and apoptosis were evaluated by SP immunohistochemical staining in 15 BLOM, 9 BLOM with dysplasia, 15 oral squamous cell carcinoma (OSCC).</p><p><b>RESULTS</b>The expression of Ki-67 in BOLM with dysplasia and OSCC was significantly higher than that of BLOM without dysplasia and normal oral mucosa (P < 0.05). The MVD in all BLOM and OSCC was significantly higher than that in normal mucosa (P < 0.05). Apoptosis in BLOM was higher than in normal mucosa and OSCC (P < 0.05).</p><p><b>CONCLUSIONS</b>The expression of Ki-67 and MVD in BLOM with dysplasia were between normal oral mucosa and oral carcinoma. The occurrence of apoptosis in BLOM was significantly higher than in normal oral mucosa. The results suggest that BLOM had the potentiality of malignant transformation.</p>

Correlation between neuronal injury and Caspase-3 after focal ischemia in human hippocampus / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cerebral ischemia is a significant clinical problem, and cerebral ischemia usually causes neuron injury such as apoptosis in various brain areas, including hippocampus. Cysteinyl aspartate-specific protease (Caspases) are fundamental factors of apoptotic mechanism. Caspase-3 inhibitors show effect in attenuating brain injury after ischemia. But all the results were from animal models in research laboratories. This study aimed at investigating the correlation between the change of ischemic neuronal injury and Caspase-3 post-ischemia in human hippocampus.</p><p><b>METHODS</b>We selected and systematized 48 post-mortem specimens from 48 patients, who died of cerebral infarction. Morphological change was firstly analyzed by observing hematoxyline/eosin-staining hippocampal sections. The expression of Caspase-3 was investigated using the methods of in situ hybridization and immunohistochemistry. Terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate-biotin nick-end labeling (TUNEL) method was used to clarify the involvement of Caspase-3 in neuron death. The loss of MAP 2 (MAP-2) was applied to judging the damaged area and degree of neuronal injury caused by ischemia.</p><p><b>RESULTS</b>In the CA1 sector of hippocampus, Caspase-3 immunostaining modestly increased at 8 hours [8.05/high-power field (hpf)], dramatically increased at 24 hours (24.85/hpf), decreased somewhat after 72 hours. Caspase-3 mRNA was detectable at 4 hours (6.75/hpf), reached a maximum at 16 hours (17.60/hpf), faded at 72 hours. TUNEL-positive cells were detectable at 24 hours (10.76/hpf), markedly increased at 48 - 72 hours. The loss of MAP-2 was obviously detected at 4 hours, progressed significantly between 24 and 72 hours; MAP-2 immunoreactivity was barely detectable at 72 hours. Before 72 hours, the Caspase-3 evolution was related with the upregulation of TUNEL and the loss of MAP-2. The positive correlation between Caspase-3 mRNA and TUNEL was significant at the 0.05 level (correlation coefficient was 0.721); the negative correlation between Caspase-3 mRNA and MAP-2 was significant at the 0.05 level (correlation coefficient is 0.857). In the early stage (before 72 hours), the staining of Caspase-3 mRNA and immunohistochemistry was predominantly present in cytoplasm; the staining of TUNEL was predominantly localized in nucleus. At 4 - 16 hours, most neurons in hippocampal CA1 areas had relatively normal morphology; at 24 - 48 hours, neurons showed apoptotic morphology; at 72 hours, most cells showed significantly pathological morphology.</p><p><b>CONCLUSIONS</b>There exist a time-dependent evolution of neuronal damage after hippocampal ischemia in human brain, which was characterized by its close correspondence to Caspase-3.</p>