A preoperative nomogram predicting risk of lymph node metastasis for early-stage cervical cancer.

OBJECTIVE: This study aimed to develop a preoperative nomogram based on clinical and pathological characteristics to provide a more individualized and accurate estimation of lymph node metastasis (LNM) in patients with early-stage cervical cancer. METHODS: A total of 7,349 early-stage cervical cancer patients with pathologically confirmed between 1988 and 2015 were obtained from the Surveillance, Epidemiology, and End Results (SEER) database. All the patients were divided into training (n = 5,500) and validation (n = 1,849) cohorts randomly. A cohort of 455 patients from multicenter was used for the external validation. We established a multivariate logistic regression model based on preoperative clinicopathological data, from which a nomogram was developed and validated. A predicted probability of LNM < 5% was defined as low risk. RESULTS: From multivariate logistic regression analysis, age at diagnosis, histologic subtype, tumor grade, tumor size and FIGO stage were identified as preoperative independent risk factors of LNM. The nomogram incorporating these factors demonstrated good discrimination and calibration (concordance index = 0.723; 95% confidence interval (CI), 0.707-0.738). In the validation cohort, the discrimination accuracy was 0.745 (95% CI, 0.720-0.770) and 0.747 (95% CI, 0.690-0.804), respectively. The nomogram was well calibrated with a high concordance probability. We also established an R-enabled Internet browser for LNM risk assessment, which tool may be convenient for physicians. CONCLUSIONS: We developed an effective preoperative nomogram based on clinical and pathological characteristics to predict LNM for early-stage cervical cancer. This model could improve clinical trial design and help physicians to decide whether to perform lymphadenectomy or not.

Transcription factors MdMYC2 and MdMYB85 interact with ester aroma synthesis gene MdAAT1 in apple.

Volatile esters in apple (Malus domestica) fruit are the critical aroma components determining apple flavor quality. While the exact molecular regulatory mechanism remains unknown, jasmonic acid (JA) plays a crucial role in stimulating the synthesis of ester aromas in apples. In our study, we investigated the effects of methyl jasmonate (MeJA) on the production of ester aroma in apples. MeJA treatment significantly increased ester aroma synthesis, accompanied by the upregulation of several genes involved in the jasmonate pathway transduction. Specifically, expression of the gene MdMYC2, which encodes a transcription factor associated with the jasmonate pathway, and the R2R3-MYB transcription factor gene MdMYB85 increased upon MeJA treatment. Furthermore, the essential gene ALCOHOL ACYLTRANSFERASE 1 (MdAAT1), encoding an enzyme responsible for ester aroma synthesis, showed increased expression levels as well. Our investigation revealed that MdMYC2 and MdMYB85 directly interacted with the promoter region of MdAAT1, thereby enhancing its transcriptional activity. In addition, MdMYC2 and MdMYB85 directly bind their promoters and activate transcription. Notably, the interaction between MdMYC2 and MdMYB85 proteins further amplified the regulatory effect of MdMYB85 on MdMYC2 and MdAAT1, as well as that of MdMYC2 on MdMYB85 and MdAAT1. Collectively, our findings elucidate the role of the gene module consisting of MdMYC2, MdMYB85, and MdAAT1 in mediating the effects of JA and promoting ester aroma synthesis in apples.

MdERF1B-MdMYC2 module integrates ethylene and jasmonic acid to regulate the biosynthesis of anthocyanin in apple.

Ethylene and jasmonic acid (JA) are crucial hormones that promote anthocyanin synthesis in apple (Malus × domestica). However, the mechanism by which these hormones cooperate to modulate anthocyanin production in apple is unclear. According to our results, MdERF1B expression was strongly induced by ethylene and JA. Physiological phenotypes and the results of molecular biological analyses indicated that MdERF1B encodes a positive regulator of anthocyanin synthesis. Specifically, MdERF1B was capable of combining directly with the MdMYC2 promoter to promote gene expression. Additionally, MdERF1B interacted with two JA signaling pathway inhibitors, namely MdJAZ5 and MdJAZ10. The MdERF1B-MdJAZ5/10 protein complex decreased the ability of MdERF1B to activate the MdMYC2 promoter. Furthermore, MdEIL1, which is a crucial protein for ethylene signal transduction, was observed to bind directly to the MdERF1B promoter, thereby upregulating gene expression. These results suggest that MdERF1B is a core gene responsive to JA and ethylene signals. The encoded protein, together with MdMYC2, MdJAZ5/10, and MdEIL1, modulates anthocyanin synthesis in apple. This study clarifies the synergistic mechanism by which JA and ethylene regulate anthocyanin production in apple.

Ethylene precisely regulates anthocyanin synthesis in apple via a module comprising MdEIL1, MdMYB1, and MdMYB17.

Ethylene regulates anthocyanin synthesis in ripening apple fruit via the antagonistic activities of the R2R3-MYB repressors and activators. However, the molecular mechanism underlying this process remains unknown. In this study, ethylene significantly induced the expression of the R2R3-MYB gene MdMYB17 in apple fruit. Moreover, MdMYB17 was revealed to be an important repressor of anthocyanin synthesis. Specifically, MdMYB17 binds directly to the promoters of the ethylene-induced genes MdMYB1 and MdEIL1, which encode positive regulators of anthocyanin synthesis, and represses their expression. Additionally, MdMYB1 and MdEIL1 bind to the MdMYB17 promoter to activate its expression. Thus, MdMYB17, MdMYB1, and MdEIL1 form a regulatory module that controls the expression of the corresponding genes. MdMYB17 interacts with MdEIL1. The interaction between MdMYB17 and MdEIL1 attenuates the regulatory effects of MdMYB17 on MdMYB1 and MdEIL1 as well as the regulatory effects of MdEIL1 on MdMYB17. Overall, our results reveal the molecular mechanisms by which MdMYB17, MdMYB1, and MdEIL1 finely mediate ethylene-regulated anthocyanin synthesis in apple fruit.

Preparation of magnetic yolk-shell structured metal-organic framework material and its application in pharmacokinetics study of alkaloids.

In this study, a magnetic yolk-shell structured metal-organic framework material (Fe3O4@YS-UiO-66-NH2) is prepared by the directional etching of Co2+/peroxymonosulfate and in situ magnetization. The characteristic properties of the material were investigated by using field emission scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectroscopy, X-ray powder diffraction, Fourier transform infrared spectroscopy, vibrating sample magnetometer, Brunauer-Emmett-Teller, and contact angle test. The Fe3O4@YS-UiO-66-NH2 shows the advantages of large surface area, good magnetic property, and satisfactory stability, as well as giving high affinity to alkaloids (ALs) via hydrophilic interaction, hydrogen bonding, and π-π interaction. The results of static adsorption experiment indicate that the Fe3O4@YS-UiO-66-NH2 possesses high adsorption capacity towards ALs and the adsorption behaviors are fitted with Langmuir adsorption isotherm model. Furthermore, a magnetic solid-phase extraction using Fe3O4@YS-UiO-66-NH2 and HPLC method was developed for the analysis of ALs in spiked samples with the recovery of 89.6-100.8%. In addition, the proposed method was successfully applied in the pharmacokinetics study of berberine, coptisine, and palmatine in the rat. In short, the developed method might be used for high-efficient recognition and determination of ALs in plasma sample, which would also provide a new way to fabricate magnetic functionalized metal-organic framework in separation science.

HE4 as a biomarker for diagnosis of lung cancer: A meta-analysis.

BACKGROUND: The aim of our study was to assess the value of serum human epididymis protein 4 (HE4) to diagnose lung cancer and provide reliable scientific conclusions to guide clinical practice. METHODS: A systematic search of the PubMed, EMBASE, Cochrane Library, Chinese National Knowledge Infrastructure, Chinese Biomedical Literature, and WANFANG databases was conducted to identify all studies examining serum HE4 in the diagnosis of lung cancer published up to June, 2017. The Quality Assessment of Diagnostic Accuracy Studies tool was used to evaluate the methodological quality of each trial. The meta-analysis was performed using STATA software and Review Manager 5.3. RESULTS: There were 21 studies involving 1883 cases and 1696 controls included in our meta-analysis. The pooled sensitivity and specificity of HE4 for diagnosing lung cancer were 0.73 (95% confidence interval [CI] 0.68-0.78) and 0.86 (95% CI 0.81-0.91), respectively. The positive likelihood ratio and negative likelihood ratio were 5.4 (95% CI 3.8-7.5) and 0.31 (95% CI 0.26-0.37), respectively. The diagnostic odds ratio was 17 (95% CI 12-26). The area under the curve of the summary receiver-operating characteristic curve was 0.86 (95% CI 0.83-0.89). Race, assay method, type of cancer, sample size, and publication date might be sources of heterogeneity in our meta-analysis. Subgroup analyses showed that the sensitivity in Caucasians was higher than that in Asians (0.81, 95% CI 0.71-0.91; and 0.71, 95% CI 0.66-0.77, respectively), but the specificity in Asians was better than that in Caucasians (0.87, 95% CI 0.81-0.92; and 0.85, 95% CI 0.73-0.97, respectively). The chemiluminescent microparticle immunoassay had the highest sensitivity, with 0.79 (95% CI 0.73-0.97), and the enzyme-linked immunosorbent assay had the highest specificity, with 0.87 (95% CI 0.79-0.94). HE4 had high diagnostic efficacy when screening for small cell lung cancer with the highest specificity (0.90, 95% CI 0.77-1.00). CONCLUSIONS: HE4 is a relatively promising and effective biomarker for the diagnosis of lung cancer. Furthermore, given the limitations of our study, additional large-scale and well-designed studies are needed in the future.

Combination of mild therapeutic hypothermia and adipose-derived stem cells for ischemic brain injury.

Mild therapeutic hypothermia has been shown to mitigate cerebral ischemia, reduce cerebral edema, and improve the prognosis of patients with cerebral ischemia. Adipose-derived stem cell-based therapy can decrease neuronal death and infiltration of inflammatory cells, exerting a neuroprotective effect. We hypothesized that the combination of mild therapeutic hypothermia and adipose-derived stem cells would be neuroprotective for treatment of stroke. A rat model of transient middle cerebral artery occlusion was established using the nylon monofilament method. Mild therapeutic hypothermia (33°C) was induced after 2 hours of ischemia. Adipose-derived stem cells were administered through the femoral vein during reperfusion. The severity of neurological dysfunction was measured by a modified Neurological Severity Score Scaling System. The area of the infarct lesion was determined by 2,3,5-triphenyltetrazolium chloride staining. Apoptotic neurons were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining. The regeneration of microvessels and changes in the glial scar were detected by immunofluorescence staining. The inflammatory responses after ischemic brain injury were evaluated by in situ staining using markers of inflammatory cells. The expression of inflammatory cytokines was measured by reverse transcription-polymerase chain reaction. Compared with mild therapeutic hypothermia or adipose-derived stem cell treatment alone, their combination substantially improved neurological deficits and decreased infarct size. They synergistically reduced the number of TUNEL-positive cells and glial fibrillary acidic protein expression, increased vascular endothelial growth factor levels, effectively reduced inflammatory cell infiltration and down-regulated the mRNA expression of the proinflammatory cytokines interleukin-1ß, tumor necrosis factor-α and interleukin-6. Our findings indicate that combined treatment is a better approach for treating stroke compared with mild therapeutic hypothermia or adipose-derived stem cells alone.

A regulatory loop involving miR-29c and Sp1 elevates the TGF-ß1 mediated epithelial-to-mesenchymal transition in lung cancer.

Specificity protein1 (Sp1) is required for TGF-ß-induced epithelial-to-mesenchymal transition (EMT) which has been demonstrated to aggravate the progression of cancer including lung cancer. microRNA-29c (miR-29c) is identified to inhibit EMT, but the correlation between miR-29c and Sp1 in human lung cancer remain incompletely clarified. Here, we confirmed decreased expression of miR-29c and enhanced expression of Sp1 in lung cancer tissues (n = 20) and found that Sp1 could be targeted and inhibited by miR-29c. Besides, the expression of miR-29c was down-regulated in high-metastatic lung cancer cell lines and TGF-ß1-treated cells. The inhibition of miR-29c or overexpression of Sp1 in 95C and A549 cells dramatically enhanced the cell migration and invasion, and also induced the decrease in the expression of epithelial markers, e.g. thyroid transcription factor 1 (TTF-1) and E-cadherin, together with an increase in mesenchymal markers including vimentin, α-smooth muscle actin (α-SMA), which could be restored by overexpression of miR-29c mimics during the TGF-ß-induced EMT. Moreover, dual-luciferase reporter assay was performed and the results indicated that miR-29c/Sp1 could form an auto-regulatory loop with TGF-ß1, which impaired TGFB1 transcription. Furthermore, miR-29c overexpression could abrogate the tumor progression and inhibit the Sp1/TGF-ß expressions in vivo, indicating that miR-29c could be a tumor suppressor and repress the Sp1/TGF-ß axis-induced EMT in lung cancer.

Heart rate variability predicts mortality in peritoneal dialysis patients.

BACKGROUND: The predictive value of heart rate variability (HRV) in peritoneal dialysis (PD) has never been tested. METHODS: In this study, the associations between HRV measures and the mortality in 81 PD patients were analyzed. HRV was measured by using 5-min recordings of a stationary system by a standardized method. Both time domain and frequency domain parameters were analyzed. RESULTS: During a follow-up period of 43.78 ± 14.77 months, 25 patients died, four patients were transferred to hemodialysis. Of the 81 patients, the time domain parameters, such as the standard deviation of differences between adjacent normal sinus to normal sinus (NN) intervals (SDSD) and the square root of the mean of the squared differences between adjacent normal NN intervals (RMSSD), were higher; the frequency domain parameters, such as the ratio of low-frequency power to high-frequency power (LF/HF) and the normalized LF, were lower, and the normalized HF was higher in the non-survived group as compared with the survived group. A Cox proportional hazards model analysis revealed that, of the HRV measures, decrease of the normalized LF, LF/HF and increase of rMSSD, SDSD, normalized HF had significant predictive value for mortality. After adjustment for other univariate predictors including age, urine volume, renal Kt/V, high-sensitivity C-reactive protein (hs-CRP), the predictive value of decreased LF/HF remained significant. Kaplan-Meier survival analysis showed mortality rate was much higher in patients with a low LF/HF (median value of 1.56). CONCLUSION: The decreases of LF/HF which reflects impaired sympathetic nerve regulation is an independent predictor of mortality in PD patients.

Heart rate variability in peritoneal dialysis patients: what is the role of residual renal function?.

BACKGROUND: The relationship between heart rate variability (HRV) and residual renal function (RRF) has not been elucidated previously. METHODS: In this cross-sectional study, HRV was evaluated in 71 peritoneal dialysis patients. Patients were divided into RRF decline group, RRF stable group and anuric group. RESULTS: RRF was negatively correlated with SDNN (r = -0.284, p = 0.017), TP (r = -0.247, p = 0.039), and HF (r = -0.238, p = 0.047). Significant sympathetic nerve activation was found in the RRF decline group (significantly lower SDNN, SDSD, RMSSD, pNN50, LF, HF, TP and higher LF/HF ratio) as compared to the RRF stable and anuric groups. Besides, significantly parasympathetic activation was found in the anuric group (the lowest LF/HF ratio as compared to the other groups (both p < 0.05). Multivariate stepwise regression analysis showed that the status of RRF was an independent factor associated with HRV parameters. CONCLUSION: This study showed autonomic nervous function in peritoneal dialysis patients was associated with a different status of RRF.

The study of spectral analysis of heart rate variability in different blood pressure types in euvolemic peritoneal dialysis patients.

BACKGROUND: Heart rate variability (HRV) is an useful noninvasive tool to assess autonomic nervous system (ANS) function and may provide an insight into the understanding of the role of ANS in the pathogenesis of blood pressure (BP) abnormality in euvolemic continuous ambulatory peritoneal dialysis (CAPD) patients. METHODS: In this cross-sectional study, 62 CAPD patients with normal hydration values (assessed by a bioimpedance spectroscopy device) were enrolled from our peritoneal dialysis (PD) unit. Patients were divided into three groups according to their BP: normotension (NT), hypertension (HT), and hypotension (HyT). Spectral analysis of HRV was measured by examination of the average heart rate using standard electrocardiogram in 5 min. RESULTS: The HyT group had the lowest serum urea nitrogen among the three groups (all p < 0.05), and a lower serum potassium and albumin as compared with the NT group (p < 0.05). The HyT group also had the highest total power (TP) and highest frequency power (HF) among the three groups (all p < 0.05), and a lower normalized low frequency power (LF), LF/HF ratio, and a higher normalized HF as compared with the HT group (all p < 0.05). CONCLUSION: Our study suggested that autonomic insufficiency was present in euvolemic PD patients, and hypotensives had a relatively higher parasympathetic activity and blunted sympathetic activity.

[Cross-sectional study of relation between blood pressure and heart rate variability in patients with peritoneal dialysis].

OBJECTIVE: To analyze the possible relationship between heart rate variability (HRV) and blood pressure in peritoneal dialysis patients. METHODS: In this cross-sectional study, 93 continuous ambulatory peritoneal dialysis patients (CAPD) patients were enrolled. Patients were divided into three groups according blood pressure. HRV which reflects autonomic nerve function (ANF) were measured by using 5-minutes recordings of a stationary system by a standardized method. Both time-domain and frequency domain parameters were analyzed. RESULTS: HRV time domain indices [the standard deviation of all normal RR intervals (SDNN), the square root of the mean of the squared differences between adjacent normal RR intervals (rMSSD), the standard deviation of differences between adjacent NN intervals (SDSD), the percentage of differences between adjacent normal RR intervals that were >50 ms computed (PNN50)] and frequency domain indices [the low-frequency energy in the power spectrum 0.04-0.15 Hz (LF), the high-frequency energy in the power spectrum 0.15-0.4 Hz (HF)] were significantly lowest in hypertensive group (P<0.05) whereas LF/HF was highest as compared to normal BP group (P<0.05). Univariable correlation analysis showed that both systolic and diastolic blood pressure were negatively correlated with SDNN (r=-0.386, P<0.001; r=-0.399, P<0.001), rMSSD (r=-0.373, P<0.001; r=-0.426, P<0.001), SDSD (r=-0.375, P<0.001; r=-0.425, P<0.001), PNN50 (r=-0.223, P=0.032; r=-0.245, P=0.018), TP (r=-0.328, P=0.001; r=- 0.312, P=0.002), LF (r=-0.260, P= 0.012; r=-0.194, P=0.063), HF (r=-0.394, P<0.001; r=-0.365, P<0.001), and positively correlated with LF/HF (r=0.275, P=0.008; r=0.171, P=0.1). Multivariable regression analysis showed that HRV indicators were independently factor affecting both systolic and diastolic blood pressure in PD patients. CONCLUSION: ANF assessed by HRV were significantly different in PD patients with blood status, parasympathetic nerve were activated in hypotension group whereas sympathetic nerve were activated in hypertension group. ANF was independently factors associated with blood pressure.

Effects of DAPT and Atoh1 overexpression on hair cell production and hair bundle orientation in cultured Organ of Corti from neonatal rats.

BACKGROUND: In mammals, hair cells do not undergo spontaneous regeneration when they are damaged and result in permanent hearing loss. Previous studies in cultured Organ of Corti dissected from neonatal animals have shown that both DAPT (r-secretase inhibitor in the Notch signal pathway) treatment and Atoh1 overexpression can induce supernumerary hair cells. The effects of simultaneous DAPT treatment and Atoh1 over expression in the cells of cultured Organ of Corti from neonatal rats are still obscure. PRINCIPAL FINDINGS: In this study, we set out to investigate the interaction of DAPT treatment and Atoh1 overexpression as well as culture time and the location of basilar fragment isolated form neonatal rat inner ear. Our results showed that DAPT treatment induced more hair cells in the apical turn, while Atoh1 overexpression induced more extra hair cells in the middle turn of the cultured Organ of Corti. When used together, their effects are additive but not synergistic. In addition, the induction of supernumerary hair cells by both DAPT and Atoh1 overexpression is dependent on the treatment time and the location of the cochlear tissue. Moreover, DAPT treatment causes dramatic changes in the orientation of the stereociliary bundles of hair cells, whereas Atoh1 overexpression didn't induce drastic change of the polarity of stereociliary bundles. CONCLUSIONS/SIGNIFICANCE: Taken together, these results suggest that DAPT treatment are much more potent in inducing supernumerary hair cells than Atoh1 overexpression and that the new hair cells mainly come from the trans-differentiation of supporting cells around hair cells. The orientation change of stereociliary bundle of hair cells may be attributed to the insertion of the newly formed hair cells. The immature hair bundles on the newly formed hair cells may also contribute to the overall chaos of the stereociliary bundle of the sensory epithelia.

The effect of blood pressure (37 vs 45 mmHg) and carotid occlusion duration (8 vs 10 min) on CA1-4 neuronal damage when using isoflurane in a global cerebral ischemia rat model.

This study presents our findings on the extent of neuronal damage in the hippocampal CA1-4 subfields following global (forebrain) cerebral ischemia in rats when using different blood pressure levels (37 vs 45 mmHg) and bilateral carotid occlusion durations (8 vs 10 min) under isoflurane anesthesia. We observed that global ischemia induced at a blood pressure of 37 mmHg resulted in high-grade CA1 neuron injury (>90%) at either duration of carotid occlusion. In contrast, global ischemia induced at a blood pressure of 45 mmHg resulted in either high-grade CA1 neuronal loss or a neuronal loss of ≈50% or less. We also noted that a post-reperfusion EEG recovery time (return of burst suppression spikes) of >12 min was associated with an 85% rate of high-grade CA1 neuronal injury. Neuronal loss in the other hippocampal subfields did not differ significantly between any of the 4 different model parameters tested. In these subfields ≈55% neuronal loss occurred in the CA2 subfield, and ≈30% in the CA3 and CA4 subfields. These findings highlight the need to assess different model parameters in order to achieve consistent high-grade CA1 neuronal damage, which, among other experimental outcomes, will improve the ability to uncover therapeutic effects using the least possible animals when assessing a neuroprotective treatment.

Comparison of the efficacy of mild hypothermia (35°c) and moderate hypothermia (33°c), alone or combined with magnesium treatment, when commenced 2 or 4 hours after global cerebral ischemia in rats.

We first assessed the neuroprotective efficacy of mild hypothermia (35°C/24 hours) alone and in combination with magnesium (intravenous loading dose: 360 µmol/kg; 48 hours infusion: 120 µmol/kg/h) commencing 4 hours after global ischemia. Treatment with mild hypothermia alone (CA1 survival rate: 8.7%±0.9%) or with magnesium (9.0%±2.9%) did not significantly increase hippocampal CA1 neuronal survival compared with saline-treated controls (7.1%±0.7%). Next, we assessed mild hypothermia (35°C/24 hours) and moderate hypothermia (33°C/24 hours), alone or in combination with magnesium, when commenced 2 hours after global ischemia. At this time point, all treatments significantly increased CA1 neuronal survival compared with saline controls (CA1 survival rates: mild hypothermia: 13.6%±1.8%; mild hypothermia + magnesium: 19.4%±8.7%; moderate hypothermia: 15.9%±4.1%; moderate hypothermia + magnesium: 21.1%±11.2%; saline control: 6.7%±1.6%; p<0.001). Although a trend for increased neuroprotection was observed when hypothermia was combined with magnesium, it did not reach statistical significance. It also appears that when a 24-hour hypothermia treatment is commenced earlier (≤2 hours postischemia) there is no difference in efficacy between mild and moderate hypothermia.

[Effect of naoxintong capsule on the vascular endothelial function and the infarct size of patients with acute myocardial infarction].

OBJECTIVE: To observe the effect of Naoxintong Capsule (NC) on the vascular endothelial function and the infarct size of patients with acute myocardial infarction (AMI). METHODS: One hundred and four patients with AMI were randomly assigned to the NC group (Group A, 36 cases), the Tongguan Capsule group (Group B, 32 cases), and the conventional Western medicine group (Group C, 36 cases). The conventional Western medicine was given to the three groups. NC was additionally given to Group A, and Tongguan Capsule was additionally given to Group B. The therapeutic course for all was 4 weeks. The plasma nitric oxide (NO), endothelin (ET), von Willebrand factor (vWF) were detected in the 3 groups before and after treatment. The inner diameter of brachial artery was examined by ultrasonograph. The flow-mediated dilation (FMD) and the nitroglycerin-mediated dilation (NMD) were calculated. The ECG QRS integral and the infarct size were assessed. RESULTS: There was no significant difference in the vascular endothelial function, ECG QRS integral, or the infarct size among the three groups before treatment (P > 0.05). Compared with before treatment, NO and NMD obviously increased after treatment in Group A and Group B, while the vWF and the infarct size obviously decreased in Group A, all showing statistical difference (P < 0.05). Compared with those in Group C, the NO, FMD, NMD significantly increased and ET obviously decreased in Group A and B after treatment (P < 0.05). The ECG QRS integral and the infarct size also decreased, with statistically significant differences in Group A (P < 0.05). Better effects on improving NO, NMD, and vWF were obtained in Group A than in Group B (P < 0.05). CONCLUSION: NC could reduce the infarct size of AMI patients possibly through improving the vascular endothelial function.

Application of real-time fluorescence quantitative PCR to rapid molecular detection of Down's syndrome.

OBJECTIVE: To develop a rapid and reliable technique for the detection of Down's syndrome. METHODS: The peripheral blood samples were collected from twenty-five Down's syndrome patients and fifty normal individuals. Four polymorphic loci on chromosomes 21, 1, 19 were amplified by real-time fluorescence quantitative PCR, and then four pairs of deltaCt values were analytically compared between the two groups. RESULTS: The deltaCt values of Down's syndrome patients were significantly lower than those of normal individuals, and the reference ranges for clinical application were primarily established. The difference between the two groups was highly significant (P < 0.001), and the reference ranges between the two groups were not overlapped. Real-time quantitative PCR technique can effectively differentiates Down's syndrome samples from the normal fetuses; furthermore, the results were consistent with those of the karyotype analysis. CONCLUSION: Real-time quantitative PCR is a fast and reliable method that may provide a new approach for rapid detection of Down's syndrome.

[Follow-up of 3 605 cord blood donors after 6 months of cord blood donation in Guangzhou cord blood bank].

To study the program of evaluating mothers and infants after 6 months of cord blood donation, from June 1998 to February 2004, all mothers after 6 months of cord blood donation were followed-up by phone calls or letters to report on the health condition. The results showed that when 3 195 mothers were visited by phone calls, 18 mothers declined to answer. 392 letter were send to those who could not be found by phone, 15 of whom wrote back. The average time to talk with each mother was approximately 12 minutes. Follow-up on the baby donors showed two cases with chromosome abnormality, one with hypothyroidism, one with neutropenia, one with albinism and 5 dead with unclear reasons. The cord blood components from all these abnormal donors found were discarded. In conclusion, the programs to evaluate mother and baby after 6 months of cord blood donation seems important in quality control of the components stored in cord blood bank.

[Effect of intraischemic mild hypothermia on interleukin-1beta and monocyte chemoattractant protein-1 contents in ischemic core of rat cortex after transient focal cerebral ischemia].

OBJECTIVE: To investigate the effect of intraischemic mild hypothermia on the protein levels of interleukin (IL)-1beta and monocyte chemoattractant protein (MCP)-1 in the ischemic core of rat cortex after transient focal cerebralischemia. METHODS: Eighty male Wistar rats were randomly divided into normothermic (37 degrees C) and mild hypothermic (32 - 33 degrees C) groups. The normothermic group was redivided into six subgroups of 8 rats: sham operation, ischemia for 2 hours without reperfusion, and reperfusion for 6 hours, 16 hours, 24 hours, and 48 hours respectively after ischemia; and the mild hypothermic group was redivided into 4 group with 8 rats: reperfusion for 6, 16, 24, and 48 hours. The rats except those in the sham operation subgroup were subjected to right middle cerebral artery occlusion by insertion a specially prepared nylon filament for two hours. Ice bag was used to lower the brain temperature and anal temperature soon after ischemia to 32.0 - 33.0 degrees C within 10 minutes in the mild hypothermic subgroups. The brain and anal temperature remained at 37.0 - 37.5 degrees C in all normothermic subgroups. Then the rats were killed 0, 6, 16, 24 and 48 hours after reperfusion respectively and their brains were taken out to examine the size of brain infarct by 2,3,5-triphenyltetrazolium chloride (TTC) staining reaction. The protein levels of IL-1beta and MCP-1 in the cortical ischemic core were measured by ELISA. RESULTS: No significant change of IL-1beta protein level was found in the cortical ischemic cores at any time point after reperfusion among the normothermic subgroups. The IL-1beta protein levels at different time points were not significantly different between the intraischemic mild hypothermia subgroups and the normothermic subgroups (all P > 0.05). The MCP-1 protein level in the cortical ischemic cores of the normothermic subgroups began to increase since the 6th hour afer reperfusion (22.5 +/- 8.7 ng x g tissue(-1), 17 times that in the sham operation samples, P < 0.05), peaked in 48 hours (110.9 +/- 47.0 ng x g tissue(-1), 83.7 times that in the sham operation sample, P < 0.001). The protein level of MCP-1 in the mild hypothermic subgroups was 8.7 +/- 7.6 ng x g tissue(-1) 6 h after reperfusion (P < 0.005 in comparison with those in sham operation subgroup and ischemia subgroup) and was 56.0 +/- 40.3 ng x g tissue(-1), 48 hours after reperfusion (P < 0.05) incomparison with those in the normothermic subgroups). The sizes of cortical infarct at different time points in the mild hypothermic subgroups were significantly smaller than those in the normothermic subgroups (P < 0.05). CONCLUSION: Mild hypothermia reduces the level of MCP-1 in the cortex after cerebral ischemia/reperfusion which may be one of the important mechanisms of the neuroprotective effects of mild hypothermia.