Multiomics and Multidimensional Testing for Efficacy Monitoring of Patients with Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To explore the role of a new blood-based, multiomics and multidimensional method for evaluating the efficacy of patients with lymphoma.@*METHODS@#10 ml peripheral blood was extracted from each patient, and the genomic copy number aberrations (CNA) and fragment size (FS) were evaluated by low-depth whole genome sequencing of cfDNA, and the level of a group of plasma tumor marker (PTM) were detected at the same time. The cancer efficacy score (CES) was obtained by standardized transformation of the value of above three numerical indexes, and the changes of CES before and after treatment were compared to evaluate the patient's response to the treatment regimen.@*RESULTS@#A total of 35 patients' baseline data were collected, of which 23 cases (65.7%) had elevated CES values. 18 patients underwent the first time test. The results showed that the CES value of 9 patients with positive baseline CES decreased significantly at the first test, and the efficacy evaluation was PR, which was highly consistent with the imaging evaluation results of the same period. At the same time, the CNA variation spectrum of all patients were evaluated and it was found that 23 patients had partial amplification or deletion of chromosome fragments. The most common amplification site was 8q24.21, which contains important oncogenes such as MYC. The most common deletion sites were 1p36.32, 4q21.23, 6q21, 6q27, 14q32.33, and tumor suppressor-related genes such as PRDM1, ATG5, AIM1, FOXO3 and HACE1 were expressed in the above regions, so these deletions may be related to the occurrence and development of lymphoma.@*CONCLUSION@#With the advantages of more convenience, sensitivity and non-invasive, this multiomics and multidimensional efficacy detection method can evaluate the tumor load of patients with lymphoma at the molecular level, and make more accurate efficacy evaluation, which is expected to serve the clinic better.

The Efficacy of Rh Phenotype Matched Blood Transfusion / 中国实验血液学杂志

OBJECTIVE@#To analyze and evaluate the efficacy of Rh phenotype matched blood transfusion.@*METHODS@#The increasing of hemoglobin (Hb) and hemolysis tests in the patients treated by Rh matched red blood cells or not, as well as the first time unmatched transfusions and the unmatched transfusions happened again after a period (≥10 d) were retrospectively analyzed.@*RESULTS@#A total of 674 times transfusions in 120 patients were evaluated. The increasing of Hb in each unit was higher in the patients treated by Rh matched blood transfusion (vs unmatched) [(33.397±1.475) g/U vs (29.951±1.304) g/U, P=0.033], while the increasing of Hb at first time unmatched transfusion and the second time unmatched transfusion was not statistically different[ (28.942±2.083) g/U vs (30.686±1.737) g/U, P=0.589]. The level of lactate dehydrogenase were related to erythrocyte washing, irradiation, period of validity and the second time unmatched transtusion (all P<0.05); the levels of total bilirubin (TBil), direct bilirubin (DBil) and indirect bilirubin (IBil) between the first time unmatched transfusion and the second time unmatched transfusion were statistically different (all P<0.05).@*CONCLUSION@#For the patients need multiple blood transfusions, Rh phenotype matched blood transfusion can reduce the exposure to Rh allogenic antigens, improve the efficacy and ensure the safety of blood transfusion.

Prospective study of the efficacy of a ketogenic diet in 20 patients with Dravet syndrome.

PURPOSE: We evaluated the efficacy and tolerability of the ketogenic diet (KD) on generalised convulsions and status epilepticus (SE) in patients with Dravet syndrome (DS). METHODS: Patients with DS having ≥2 generalised convulsions/month despite drug treatment were included in this study and placed on a KD for 6 months. From 3 months before (baseline) to 6 months after KD initiation, caregivers recorded patients' seizure activity, antiepileptic drug use, and adverse events. The KD efficacy was determined by examining the frequency and duration of seizures at 3 and 6 months vs. baseline. Responders were defined as individuals whose generalised convulsions decreased in frequency by ≥50% vs. baseline. Seizures lasting ≥5 min and SE were specifically evaluated. Patients' cognition was also assessed at 3 and 6 months via questionnaire. RESULTS: Twenty patients continued the KD for at least 3 months. Of the 17 responders identified at month 3, seizures decreased by 50-89% and 90-99% in nine and two patients, respectively; six patients were seizure free. The KD was ineffective in three patients, who discontinued the diet. By month 6, seizures decreased by 50-89% and 90-99% in six and one patient(s), respectively; 10 patients were seizure free. The frequency of other seizure types also improved. During all 6 months, neither generalised convulsions lasting ≥5 min nor SE was detected in the 17 responders. The KD also improved patients' cognition. CONCLUSION: The KD is a good treatment option for medically intractable epilepsy.

Correlation between cystatin C and atherogenic index of plasma change in hypertensive patients / 预防医学

Objective To understand the correlation and clinical significance between cystatin C and atherogenic index of plasma change in hypertensive patients. Methods At the Quzhou City central hospital between 2014 and 2015, 526 cases of hypertensive patients as hypertensive group and 546 cases of people with normal blood pressure in the healthy check-up as normal blood pressure group were investigated with physical examination, blood biochemical index detection and the serum cystatin C level detection. The analysis of the relationship between the serum cystatin C level and atherogenic index of plasma among two groups was done. Results The evidence that the serum cystatin C level between hypertensive group and normal blood pressure group shows respectively as: 1.12±0.44 (mg/L) and 0.81±0.22 (mg/L), atherogenic index of plasma shows respectively as:0.68±0.03 and -0.22±0.02, both differences were statistically significant (P<0.01) . As the serum cystatin C level increased, the risk of hypertension increased (OR=20.06, 95％CI: 12.67-31.76) . Plasma arteriosclerosis index in hypertensive group was correlated with systolic blood pressure, body mass index, total cholesterol, triglyceride, high-density lipoprotein, LDL cholesterol, and uric acid level respectively, all differences were statistically significant. In addition to the above indicators, the serum cystatin C level in hypertensive group was correlated with serum creatinine level (all P<0.05) . Conclusion The serum cystatin C level and plasma arteriosclerosis index in patients with hypertension both were higher than those with normal blood pressure. These two indicators were correlated with systolic blood pressure and multiple blood lipid indicators. They could be used to monitor arteriosclerosis and target organ damage in patients with hypertension.

Effects of Different Storage Time and Temperature on Coagulation Function of Fresh Frozen Plasma after Thawing / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the coagulation function changes of fresh frozen plasma storaged at different storage time and temperature after thawing.</p><p><b>METHODS</b>Forty unit of fresh frozen plasma were collected, and thawed at 37 °C for 25 minutes. Each unit was divided into 2 halves: one was stored at (4±2)°C for 72 hours and the other one was stored at (25±2) °C for 72 hours. At the time point of 0, 4, 24, 48 and 72 h, thromboela-stogram TEG was measured for all samples. At the the same time, factors V,VII,VIII and IX, APTT and PT were also measured for all the samples. Blood culture for all the samples was used to discover aerobic and anaerobic bacteria.</p><p><b>RESULTS</b>All the samples could form a stable blood clot after thawing for 72 h, and the blood culture results of all samples were negative. Significant changes were observed in ACT and TMA between 0 h and other test time, but there was no difference between 4 °C and 25 °C. The activity of factor V was significantly different between 4 °C and 25 °C after storing for 48 and 72 hours, which was reduced faster at 25 °C.</p><p><b>CONCLUSION</b>Although part of the coagulation factor activity were attenuated after fresh frozen plasma being thawed and stored for 72 hours at different temperature, but all samples can form stable blood clots. Fresh frozen plasma stored for more than 72 hrs after thawing can be used to supplement the coagulation factors to patient.</p>

Dynamic changes of pH2AX expression in the reversibility of mouse testicular reproductive function impaired by single heat stress / 中华男科学杂志

Objective@#To investigate the role of pH2AX in the reversibility of mouse testicular reproductive function impaired by single heat stress.@*METHODS@#Twenty-four C57 male mice were randomly divided into heat stress and control groups and immersed in water at 43℃ and 25℃, respectively, for 15 minutes. At 1, 7, and 14 days of heat exposure, all the mice were sacrificed and their testis tissues collected for determining the apoptosis of the germ cells by TUNEL and measuring the expression level of the pH2AX protein by immunohistochemistry and Western blot.@*RESULTS@#The highest percentage of apoptotic cells were found in the seminiferous tubules of the mice in the heat stress group on the 1st day of the exposure and almost no apoptosis was observed at 7 and 14 days. The pH2AX protein was expressed in the nuclei of the basement membrane of adjacent seminiferous tubules. Compared with the control group, the expression of pH2AX was significantly increased on the 1st day of exposure (0.47 ± 0.02 vs 1.61 ± 0.04, P <0.01), then decreased at 7 days (0.85 ± 0.03) in comparison with that on the 1st day (P <0.01), and again elevated at 14 days (1.72 ± 0.02) as compared with either those at 1 and 7 days (P <0.01) or that of the control (P <0.01).@*CONCLUSIONS@#Heat stress causes dynamic changes of the pH2AX expression in the testis of the mouse, which are associated with heat stress-induced proliferation and division of the testicular spermatogenic cells.

Haoqin Qingdan Decoction () and ribavirin therapy downregulate CD14 and toll-like receptor 4 in febrile disease with dampness-heat syndrome in a mouse model / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of Chinese medicine Haoqin Qingdan Decoction (, HQD) for febrile disease dampness-heat syndrome (FDDHS).</p><p><b>METHODS</b>Forty mice were divided into four groups, including normal control, FDDHS (induced by Radix et Rhizoma Rhei recipe and influenza virus A1 FM1 model), HQD, and the ribavirin groups (10 in each). The normal control and FDDHS groups were administered normal saline. HQD and the ribavirin groups were administered HQD and ribavirin intragastrically once daily at a dose of 64 g/(kg d) and 0.07 g/(kg d), respectively for 7 days. Lethargy, rough hair, diarrhea, tongue color and sole color were evaluated for pathological changes in morphology. The tongue and lung tissues were collected for histology. The CD14 and toll-like receptor 4 (TLR4) expression levels were measured using real-time quantitative polymerase chain reaction.</p><p><b>RESULTS</b>More than 80% of the FDDHS mice showed hypokinesia and lethargy, and pathological changes associated with rough hair, diarrhea, tongue color and sole color. With advanced treatment for 7 days, the thick greasy tongue fur of the HQD and ribavirin groups were thinner than that of the FDDHS group (P<0.05), and it was the thinnest in the ribavirin group as compared with that in other groups (P<0.05). The CD14 and TLR4 expression levels in the lung tissues of HQD and ribavirin groups significantly delined compared with the model group (P<0.05 or P<0.01). CD14 was down-regulated more remarkably in the HQD group compared with the ribavirin group (P<0.05), whereas the converse was true with TLR4 (P<0.05).</p><p><b>CONCLUSIONS</b>We established a FDDHS mouse model showing systemic clinical symptoms. Both HQD and ribavirin can inhibit the expression of CD14 and TLR4 in FDDHS mice, while the effect of ribavirin might be much more violent. The expression changes of CD14 and TLR4 consistently refers to lipopolysaccharide, the commonly and hotly inducing factor in FDDHS.</p>

Clinical Significance of Detecting CyclinD1 and BCL-2 in Patients with B-Cell Lymphoma by Using Flow Cytometry / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the feasibility and value of detecting CyclinD1 and BCL-2 in patients with B-cell lymphoma by using flow cytometry.</p><p><b>METHODS</b>Fifty-three patients with lymphoma were selected, and 50 healthy persons in the same period were selected as control. The expression levels of CyclinD1 and BCL-2 in patients with various subtypes of lymphoma were detected by using flow cytometry (FCM).</p><p><b>RESULTS</b>When the dilution time was 1 min and the dilution proportion was 1:20, the cell morphology was the best complete, at the 4 min the cell morphology was best status. The mean fluorescence intensity of CyclinD1 and BCL-2 between persons of control group and patients with B-cell lymphoma showed significant difference, the CyclinD1 level (1.824 ± 0.315) and BCL-2 levels (4.257 ± 0.528) of patients with B-cell lymphoma were obviously higher than the CyclinD1 level (0.634 ± 0.153) and BCL-2 level (1.926 ± 0.328) of persons in control group, the CyclinD1 and BCL-2 expression levels of patients with HL were significantly lower than CyclinD1 and BCL-2 levels of patients with NHL (P < 0.01). After treatment, the expression levels of CyclinD1 and BCL-2 in patients with B lymphoma were significantly lower than these befor treatment.</p><p><b>CONCLUSION</b>Using the method of flow cytometry for detecting CyclinD1 and BCL-2 expression levels in lymphoma cells of patients is feasible, and it can be applied clinically to evaluate the treatment efficacy.</p>

Effect of Yunnan herb Laggera pterodonta against influenza A (H1N1) virus in vitro / 中国中药杂志

Laggera pterodonta is commonly used for treating influenza in Southwest China, especially in Yunnnan province. The main clinical effects of L. pterodonta include anti-influenza, anti-microbial, anti-inflammatory. To investigate the anti-influenza A (H1N1) virus effect of L. pterodonta, neutralization inhibition and proliferation inhibition tests were performed. MDCK culture method was used to observe the cytopathic effect (CPE) of extracts from L. pterodonta in inhibiting influenza A (H1N1) virus and haemagglutination titre of H1N1 virus in vitro. The culture medium were collected at 24 h, 48 h, 72 h, 96 h, and detected by Real time RT-PCR, in order to compare the effect of different extracts from L. pterodonta on in vitro proliferation of H1N1, virus. The result of neutralization inhibition test showed that hemagglutination titer of ethyl acetate extract were 8 times lower at 72 h; in proliferation inhibition test, hemagglutination titer of ethyl acetate extracts reduced by 2 and 4 times. According to the results of Real time RT-PCR test, the H1N1 inhibition ratio of ethyl acetate extract was 72.5%, while the proliferation inhibition ratio of ethyl acetate extract was 25.3%; as for petroleum ether extracts, the H1N1 inhibition ratio was 60.2%, while the proliferation inhibition ratio was 81.4%. In conclusion, both ethyl acetate extract and petroleum ether extract of L. pterodonta have significant neutralization and direct proliferation inhibition effects on influenza A virus.

Mechanism of cooked blanched garlic leaves against platelet aggregation / 中国实验血液学杂志

This study was purposed to explore the mechanism of cooked blanched garlic leave juice against platelet aggregation. The juice of blanched garlic leaves was mixed with platelet rich plasma (PRP), the human platelet aggregation, the activation of human platelets induced by adenosine diphosphate (ADP) and collagen were observed; the expression levels of the activated platelets (Fib-R) and P-selectin (CD62P), and the amount of platelet fibrinogen binding were detected by flow cytometry; 10 rabbits were randomly divided into two groups, in addition to the normal diet, they were fed with physiologic saline and cooked blanched garlic leave juice respectively. After 1, 3, 5 , 8 weeks, the maximum ratio of rabbit platelet aggregation induced by ADP and collagen were observed . The results showed that the cooked blanched garlic leave juice could significantly inhibit human platelet aggregation induced by ADP and collagen (P < 0.05), the inhibitory ratio were 87.37% and 86.24% respectively; the juice could not inhibit activated platelets Fib-R and CD62P expression levels (P > 0.05), but was able to inhibit platelet fibrinogen binding capacity (P < 0.05); the rabbit platelet aggregation rate in the group given cooked blanched garlic leave juice was significantly lower than that in control group (P < 0.05). It is concluded that the cooked blanched garlic leave juice can inhibit platelet aggregation in vitro and in vivo, the inhibition of aggregation pathway mainly is blocking the combination of fibrinogen with Fib-R, which finally results in the inhibition of platelet aggregation. Therefore, regular consumption of cooked blanched garlic leaves may prevent cardiovascular thrombotic diseases.

Analysis on genotype distributions and epidemiological characteristics of Yersinia pestis plague foci in Gansu province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the genotype distributions and epidemiological characteristics of Yersinia pestis in Gansu province.</p><p><b>METHODS</b>Primers were designed according to the confirmed 23 differential sections, to genotype the 202 Yersinia pestis DNA of Gansu province by PCR, and to analyze its distribution and epidemiological characteristics.</p><p><b>RESULTS</b>Yersinia pestis in Gansu province could be divided into eight genotypes: 1b, 5, 7, 8, 13, 26, new genotype 1 (GS1) and new genotype 2 (GS2). They were distributed in various regions. 1b, 8 and GS1 genotypes of Yersinia pestis had been identified since 1960s but the 7, 13 and 26 genotypes had not been isolated for more than 40 years while GS2 and 5 genotypes had been isolated since 1990s.</p><p><b>CONCLUSION</b>1b, 8 and GS1 genotypes of Yersinia pestis continued to be violently prevalent since 1960s but 7, 13 and 26 genotypes had not been isolated for more than 40 years while GS2 and 5 genotypes had started to be popular since 1990s.</p>

Left atrial wall thickness: anatomic aspects relevant to catheter ablation of atrial fibrillation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The success and complication rates of atrial fibrillation (AF) ablation may be related to regional differences in left atrial (LA) wall thickness. The purpose of this study was to investigate the transmural LA wall thickness in various regions.</p><p><b>METHODS</b>We measured LA wall thickness in 36 human heart specimens using calipers at three planes including left pulmonary veins (PVs) vestibule plane, right PVs vestibule plane and the middle plane between the two. In each plane, eight points were selected, including superior, middle and inferior levels at anterior and posterior wall, roof and bottom.</p><p><b>RESULTS</b>The anterior and posterior wall thickness displayed gradient from superior to inferior level (anterior wall: (2.73 ± 1.01) mm, (2.08 ± 0.91) mm and (1.54 ± 0.69) mm; posterior wall: (1.74 ± 0.68) mm, (1.48 ± 0.39) mm and (1.27 ± 0.42) mm). At the roof, LA wall thickness was thickest in middle plane ((2.01 ± 1.02) mm) and was thinnest in left PVs vestibule plane ((1.29 ± 0.41) mm). The posterior wall thickness in left PVs vestibule plane was thinner than in the other two planes (P < 0.05 - 0.001), and was thinner in right PVs vestibule plane than in middle plane (P < 0.01 - 0.001). Whereas in anterior wall, the wall thickness in left PVs vestibule plane was thicker than in middle and right PVs vestibule plane.</p><p><b>CONCLUSIONS</b>Significant variations exist for mean LA wall thickness at different regions which are often targeted during circumferential pulmonary venous ablation (CPVA). Appreciating these differences may have significant implications in catheter ablation of AF.</p>

Dynamic prediction on the number of influenza-like cases in Gansu province based on data from the influenza sentinel surveillance program, from 2006 to 2011 / 中华流行病学杂志

Objective To understand the epidemiological trend on the number of influenzalike cases and to explore the feasibility of early warning systems of influenza in Gansu province.Methods Based on data from the influenza sentinel surveillance program,a sequence chart was used to analyze the epidemiological trend on the number of influenza-like illness (ILI) cases.Both control chart and mobile percentile method were used to select the threshold of premium alert for the ILI of sentinel surveillance program.Warning effects were assessed by statistical model.Results The prevalence of influenza were both low in 2007 and 2008.Alert thresholds for ILI of Sentinel surveillance was built.The thresholds were higher alert in winter,but lower in summer.Both Seasonal Exponential Smoothing Model and Multiplicative Seasonal ARMA Model (1,1,1) (0,1,0) were used to dynamically predict the weekly percentage of outpatient visits for influenza-like illness (ILI％)of 2011.The concordance rates (predicted=actual) were 100％ for both of them.According to the RMSE values,the dynamically predicted effect of the seasonal exponential smoothing model was superior to ARIMA.Conclusion Dynamic prediction on the number of influenza-like cases could reflect the epidemiological trend of influenza in Gansu province,but with some limitations.

Cyclooxygenase inhibitors in some dietary vegetables inhibit platelet aggregation function induced by arachidonic acid / 中国实验血液学杂志

The study was purposed to investigate whether the cyclooxygenase inhibitors from some dietary vegetables can inhibit platelet aggregation function by the arachidonic acid (AA). The vegetable juice was mixed with platelet rich plasma (PRP), and asprin was used as positive control. The maximum ratio of platelet aggregation induced by AA was measured on the aggregometer; heme and cyclooxygenase-1 (COX(1)) or cyclooxygenase-2 (COX(2)) were added to test tubes containing COX reaction buffer, the mixture was vortex-mixed and exposed to aspirin or vegetable juice, followed by addition of AA and then hydrochloric acid (1 mol/L) was added to stop the COX reaction, followed by chemical reduction with stannous chloride solution. The concentration of COX inhibitors was detected by the enzyme immunoassay kit; vegetable juice (aspirin as positive control) was mixed with whole blood, which was followed by the addition of AA, and then the reaction was stopped by adding indomethacin, centrifuged, then the supernatant was collected, and the plasma thromboxane B(2) (TXB(2)) was measured by radioimmunoassay. The results showed that spinach juice, garlic bolt juice, blanched garlic leave juice and Chinese leek juice could inhibit by 80% human platelet aggregation induced by AA. 4 kinds of vegetables were all found a certain amount of cyclooxygenase inhibitors, which COX(1) and COX(2) inhibitor concentrations of spinach were higher than that of aspirin; 4 vegetable juice could significantly reduce the human plasma concentrations of TXB(2) induced by AA (p < 0.05). It is concluded that 4 kinds of raw vegetables containing cyclooxygenase inhibitors inhibit the production of TXA(2) and thus hinder platelet aggregation. Raw spinach, garlic bolt, blanched garlic and chinese leek inhibit significantly AA-induced human platelet aggregation in vitro. 4 kinds of vegetables may have a good potential perspective of anti-platelet aggregation therapy or prevention of thrombosis.

Expression of S100A4 in esophageal squamous cell carcinoma and its relation to tumor invasion and metastasis / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the role of S100A4 in the carcinogenesis, development, invasion and metastasis of esophageal squamous cell carcinoma.</p><p><b>METHODS</b>Immunohistochemistry was used to detect the expressions of S100A4, MMP-2 and E-cadherin proteins in 100 cases of surgically resected esophageal squamous cell carcinoma specimens. RT-PCR and Western blot were used to detect the expressions of S100A4 mRNA and protein in esophageal squamous cell carcinoma line EC-1 and TE-1. Boyden-chamber model in vitro was utilized to detect the invasion ability of EC-1 and TE-1 cells.</p><p><b>RESULTS</b>The positivity rate of S100A4 protein was 52.0% was in esophageal carcinoma tissues, significantly higher than that in normal tissues (26.0%) (P<0.01). The expression of S100A4 was related to tumor grading, invasive depth and lymph node metastasis (P<0.05). In esophageal carcinoma, the expression of S100A4 was positively correlated to MMP-2 expression (P<0.01), but inversely to E-cadherin expression (P<0.05). The expressions of S100A4 mRNA (0.894-/+0.021) and protein (0.897-/+0.053) in EC-1 cells were significantly higher than those in TE-1 (0.812-/+0.040 and 0.645-/+0.089, respectively, P<0.01), and the invasion ability of EC-1 cells was significantly higher than that of TE-1 cells (91.00-/+17.44 vs 61.80-/+11.10, P<0.01).</p><p><b>CONCLUSION</b>The overexpression of S100A4 in esophageal squamous cell carcinoma tissue and highly invasive EC-1 cells may contribute to the carcinogenesis, development, invasion and metastasis of esophageal squamous cell carcinoma.</p>

RNA interference targeting mu-opioid receptors reverses the inhibition of fentanyl on glucose-evoked insulin release of rat islets / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mu opioid receptor plays an important role in many physiological functions. Fentanyl is a widely used opioid receptor agonist for analgesia. This study was conducted to test the role of mu-opioid receptor on insulin release by determining whether fentanyl affected insulin release from freshly isolated rat pancreatic islets and if small interfering RNAs (siRNA) targeting mu-opioid receptor in the islets could knock down mu-opioid receptor expression.</p><p><b>METHODS</b>Islets were isolated from ripe SD rats' pancreas by common bile duct intraductal collagenase V digestion and purified by discontinuous Ficoll density gradient centrifugation. The siRNA knock-down of mu-opioid receptor mRNA and protein in islet cells was analyzed by semi-quantitative real time-PCR and Western blotting. After siRNA-transfection for 48 hours, the islets were co-cultured with fentanyl as follows: 0 ng/ml, 3 ng/ml and 30 ng/ml for 48 hours. Then glucose-evoked insulin release was performed. As a control, the insulin release was also analyzed in islets without siRNA-trasfection after being co-cultured with fentanyl for 48 hours.</p><p><b>RESULTS</b>After 48 hours of transfections, specific siRNA targeting of mu-opioid receptors produced significant reduction of mu-opioid receptor mRNA and protein (P < 0.01). Fentanyl significantly inhibited glucose-evoked insulin release in islets in a concentration dependent manner (P < 0.01). But after siRNA-transfection for 48 hours, the inhibition on glucose-evoked insulin release was reversed (P < 0.01).</p><p><b>CONCLUSIONS</b>RNA interference specifically reduces mu-opioid receptor mRNA and protein expression, leading to reversal of the fentanyl-induced inhibition on glucose-evoked insulin release of rat islets. The activation of opioid receptor induced by fentanyl functions to inhibit insulin release. The use of RNAi presents a promising tool for future research in diabetic mechanisms and a novel therapy for diabetes.</p>

Clinical analysis and screening for SCN1A gene mutation in two pedigrees of generalized epilepsies with febrile seizures plus / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the clinical and genetic characteristics of generalized epilepsy with febrile seizures plus (GEFS).</p><p><b>METHODS</b>Data of two probands of the disease were collected through outpatient clinic. DNA was extracted from peripheral blood leukocytes using RelaxGene Blood DNA System. Twenty-six exons of SCN1A were amplified by polymerase chain reaction (PCR), the PCR products were screened by denaturing high performance liquid chromatography (DHPLC), then the abnormal fragments were sequenced by Sanger method in order to find the mutations of SCNIA gene.</p><p><b>RESULTS</b>(1) There were 28 affected individuals in the two families of GEFS+ (14 males and 14 females). Febrile seizures (FS) were present in 7 cases, febrile seizures plus (FS+) in 6 cases, FS+ and absence seizures in 1 case, FS+ and myoclonic seizures in 1 case, uncertain type in 13 cases. No severe phenotype was seen. Bilineal inheritance occured in one GEFS+ family. (2) A samesense mutation (c. 1212A > G) of SCN1A gene was found in the proband and an unaffected individual of pedigree B of GEFS.</p><p><b>CONCLUSIONS</b>(1) GEFS+ is a syndrome with the characteristics of heterogeneous clinical phenotypes; bilineal inheritance suggests the possibility of complex inheritance with additive gene effects. (2) Our study failed to provide evidence supporting a causal relation between the SCN1A mutation and the etiologic gene in the GEFS+ family B, which indicates that GEFS+ has the phenotypic and genotypic heterogeneity.</p>

Abnormal expression and significance of Runx2 in osteoblasts of adolescent idiopathic scoliosis patients / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the possible relationship between Runx2 and the low bone mass of adolescent idiopathic scoliosis(AIS) patients at the osteoblast level.</p><p><b>METHODS</b>Twenty eight AIS patients (mean age 14.9 years, mean Cobb angle 57.3 degrees ) in experimental group, including 2 male and 26 female, underwent posterior instrumentation between March and December 2008. They were divided into two groups. Patients in group A maintained normal bone mineral density (BMD). Patients in group B sustained osteopenia. Normal group, including 8 patients (2 males and 6 females) with a mean age of 15.3 years, were age-matched non-scoliosis adolescents who underwent spinal surgery. BMD of the lumbar spine and proximal femur was measured by using dual energy X-ray absorptiometry in three groups. Small cancellous bone samples were harvested from the iliac crest during the operation. The chipped explants were cultured to obtained the osteoblasts. P2 generation osteoblasts were analyzed to confirm the cell phenotype. Expression of mRNA and protein of Runx2 were detected by using RT-PCR and Western blot in P2 generation osteoblasts from three groups.</p><p><b>RESULTS</b>The expression of Runx2 of osteoblasts had decreased obviously in group B compared with group A and group C (P < 0.05). However, there was no significant difference between group A and group C (P > 0.05).</p><p><b>CONCLUSIONS</b>The abnormal expression of Runx2 of osteoblasts may be responsible for the low bone mass in adolescent idiopathic scoliosis patients.</p>

Association between insulin-like growth factor-1 receptor gene polymorphisms and with susceptibility to adolescent idiopathic scoliosis / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the association of insulin-like growth factor-1 receptor (IGF-1R) gene polymorphisms and with susceptibility to adolescent idiopathic scoliosis (AIS).</p><p><b>METHODS</b>This study included 200 patients with AIS and 200 healthy controls. Height, menarche status, curve pattern, Cobb angle, and Risser sign in female patients were recorded. Blood samples were taken form each subject by venipuncture. Genetic DNA was extract from peripheral blood leukocytes using standard phenol/chloroform extraction. PCR-RFLP (polymerase chain reaction-restriction-fragment length polymorphism) was used for the genotyping.</p><p><b>RESULT</b>The genotype and allele frequency distribution were similar between AIS and normal control (P>0.05). The mean maximum Cobb angle, Risser sign, menarche status of different genotypes of IGF-1R gene were similar with each other among female AIS patients (P>0.05).</p><p><b>CONCLUSIONS</b>The IGF-1R gene is neither associated with the occurrence nor the curve severity of AIS.</p>

Transforming growth factor beta1 regulation of epithelial-mesenchymal transition in esophagus squamous cell carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the functional role of transforming growth factor beta1(TGFbeta1) in the regulation of epithelial-mesenchymal transition (EMT) and the effect of TGFbeta1-ASODN blockage of EMT in esophagus squamous cell carcinoma.</p><p><b>METHODS</b>Esophageal squamous cell carcinoma cell line EC9706 was transfected with chemically synthesized TGFbeta1-ASODN. RT-PCR, immunohistochemistry and flow cytometry were used to detect the protein and mRNA expressions of TGF-beta1, E-cadherin and vimentin before and after the transfection. Morphological changes were documented and scarification test was used to detect the migration potential of EC9706 before and after the transfection.</p><p><b>RESULTS</b>After TGFbeta1-ASODN transfection, mRNA (0.25 +/- 0.07) and protein (35.07% +/- 1.42%) expressions of TGFbeta1 in EC9706 were significantly lower than those before transfection (mRNA: 0.43 +/- 0.09; protein: 43.57% +/- 1.77%, chi(2) = 13.847 and chi(2) = 84.120, P < 0.05). The mRNA (0.38 +/- 0.09) and protein (17.13% +/- 1.45%) expressions of E-cadherin were significantly higher than those before transfection (0.22 +/- 0.06; 12.53% +/- 1.31%, chi(2) = 0.160 and chi(2) = 40.008, P < 0.05) and the mRNA (0.73 +/- 0.07) and protein (14.15% +/- 1.46%) expressions of vimentin were significantly lower than those (0.89 +/- 0.09; 17.97% +/- 1.42%) before transfection (chi(2) = 0.160 and chi(2) = 21.103, P < 0.05). Scarification test showed that after transfection, the mobility of EC9706 was significantly inhibited and its migration length (0.45 +/- 0.05) was significantly shorter than that before the transfection (0.81 +/- 0.11, chi(2) = 16.854, P < 0.05).</p><p><b>CONCLUSIONS</b>TGFbeta1 may contribute to EMT in esophageal squamous cell carcinoma. TGFbeta1-ASODN leads to an over-expression of E-cadherin and a down-regulation of vimentin, along with the morphological alterations and migration inhibition, indicating that a blockage of TGFbeta1 suppresses EMT in esophagus squamous cell carcinoma.</p>