Different effects of long-term and short-term repeated restraints on the hematopoietic stem cells in mice / 生理学报

Humans with chronic psychological stress are prone to develop multiple disorders of body function including impairment of immune system. Chronic psychological stress has been reported to have negative effects on body immune system. However, the underlying mechanisms have not been clearly demonstrated. All immune cells are derived from hematopoietic stem cells (HSC) in the bone marrow, including myeloid cells which comprise the innate immunity as a pivotal component. In this study, to explore the effects of chronic psychological stress on HSC and myeloid cells, different repeated restraint sessions were applied, including long-term mild restraint in which mice were individually subjected to a 2 h restraint session twice daily (morning and afternoon/between 9:00 and 17:00) for 4 weeks, and short-term vigorous restraint in which mice were individually subjected to a 16 h restraint session (from 17:00 to 9:00 next day) for 5 days. At the end of restraint, mice were sacrificed and the total cell numbers in the bone marrow and peripheral blood were measured by cell counting. The proportions and absolute numbers of HSC (LinCD117Sca1CD150CD48) and myeloid cells (CD11bLy6C) were detected by fluorescence activated cell sorting (FACS) analysis. Proliferation of HSC was measured by BrdU incorporation assay. The results indicated that the absolute number of HSC was increased upon long-term mild restraint, but was decreased upon short-term vigorous restraint with impaired proliferation. Both long-term mild restraint and short-term vigorous restraint led to the accumulation of CD11bLy6C cells in the bone marrow as well as in the peripheral blood, as indicated by the absolute cell numbers. Taken together, long-term chronic stress led to increased ratio and absolute number of HSC in mice, while short-term stress had opposite effects, which suggests that stress-induced accumulation of CD11bLy6C myeloid cells might not result from increased number of HSC.

Correlation of genes expression on MyD 88-dependent signaling pathway in progression of knee osteoarthritis / 中国骨伤

<p><b>OBJECTIVE</b>To investigate expression features and correlation of genes expression on MyD88-dependent signaling pathway in synovial membrane (SM) of progression of knee osteoarthritis (OA).</p><p><b>METHODS</b>Sixty Wistar rats were randomly divided into 6 groups, including blank group (N), false surgical group, model groups[2 weeks (2W), 4 weeks (4W), 8 weeks (8W) and 12 weeks (12W)], with 10 rats in each group. The models were established by using Hulth method. Control group was experienced no surgery, while false surgical group was only opened joint cavity and sutured. The SM samples was collected according to the time designed above. The relative expression quantity of MyD88, TLR4 and NF-κB was detected by Real-time PCR after the extraction of the total RNA and reverse transcription. The correlation analysis was obtained by SPSS.</p><p><b>RESULTS</b>There was no significant difference in each gene mRNA expression between false surgical and blank group(> 0.05), while enhanced expression was found in the model groups(<0.05). The correlation index among MyD88, TLR4 and NF-κB was 0.91 and 0.86 respectively, and had significant difference among them.</p><p><b>CONCLUSIONS</b>Positively relative among MyD88, TLR4 and NF-κB played main role in TLR4/NF-κB signal passway, and could predicate the expression of other genes in the passway. It also could further provide the basis for clarify the pathologic mechanism of knee OA.</p>

Effects of Risperidone and Paliperidone on Brain-Derived Neurotrophic Factor and N400 in First-Episode Schizophrenia / 中华医学杂志(英文版)

<p><b>Background</b>Risperidone and paliperidone have been the mainstay treatment for schizophrenia and their potential role in neuroprotection could be associated with brain-derived neurotrophic factor (BDNF) and N400 (an event-related brain potential component). So far, different effects on both BDNF and N400 were reported in relation to various antipsychotic treatments. However, few studies have been conducted on the mechanism of risperidone and paliperidone on BDNF and N400. This study aimed to compare the effects of risperidone and paliperidone on BDNF and the N400 component of the event-related brain potential in patients with first-episode schizophrenia.</p><p><b>Methods</b>Ninety-eight patients with first-episode schizophrenia were randomly divided into the risperidone and paliperidone groups and treated with risperidone and paliperidone, respectively, for 12 weeks. Serum BDNF level, the latency, and amplitude of the N400 event-related potential before and after the treatment and Positive and Negative Syndrome Scale (PANSS) scores were compared between the two groups.</p><p><b>Results</b>A total of 94 patients were included in the final analysis (47 patients in each group). After the treatment, the serum BDNF levels in both groups increased (all P < 0.01), while no significant difference in serum BDNF level was found between the groups before and after the treatment (all P > 0.05). After the treatment, N400 amplitudes were increased (from 4.73 ± 2.86 μv and 4.51 ± 4.63 μv to 5.35 ± 4.18 μv and 5.52 ± 3.08 μv, respectively) under congruent condition in both risperidone and paliperidone groups (all P < 0.01). Under incongruent conditions, the N400 latencies were shortened in the paliperidone group (from 424.13 ± 110.42 ms to 4.7.41 ± 154.59 ms, P < 0.05), and the N400 amplitudes were increased in the risperidone group (from 5.80 ± 3.50 μv to 7.17 ± 5.51 μv, P < 0.01). After treatment, the total PANSS score in both groups decreased significantly (all P < 0.01), but the difference between the groups was not significant (P > 0.05). A negative correlation between the reduction rate of the PANSS score and the increase in serum BDNF level after the treatment was found in the paliperidone group but not in the risperidone group.</p><p><b>Conclusions</b>Both risperidone and paliperidone could increase the serum BDNF levels in patients with first-episode schizophrenia and improve their cognitive function (N400 latency and amplitude), but their antipsychotic mechanisms might differ.</p>

Effects of Short-term High Dose Atorvastatin on Left Ventricular Remodeling in Patients with First Time Attack of Anterior Acute Myocardial Infarction / 中国医学科学杂志(英文版)

Objects The aim of this trial was to evaluate the effect of short-term high-dose atorvastatin therapy on levels of high-sensitivity C-reactive protein (hs-CRP), malonaldehyde (MDA), endothelin-1(ET-1), matrix metalloproteinases (MMPs), and left ventricular (LV) remodeling in patients with first time attack of acute anterior myocardial infarction (AAMI) .Methods A hundred and three patients with first time attack of AAMI who underwent successful primary percutaneous coronary intervention were randomized to receive atorvastatin 40 mg once daily for 1 week followed by 20 mg once daily (intensive treatment group, IT group, n=49), or atorvastatin 20 mg once daily (standard treatment group, ST group, n=54). Plasma levels of hs-CRP, MDA, ET-1, MMP-2 and MMP-9 were measured on admission, at 1 week, 2 weeks and 6 months follow up and compared between the IT group and ST group. Echocardiography was performed on admission, at 2 week, and 1 year follow up. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV) and left ventricular ejection fraction (LVEF) were measured at each echocardiographic examination and compared between the IT group and ST group.Results Plasma levels of hs-CRP (F=7.718, P=0.009), ET-1 (F=7.882, P=0.006), MMP-9 (F=4.834, P=0.028) and pro-BNP (F=4.603, P=0.032) were significantly lower at 1 week after initial onset of AAMI in the IT group compared with the ST group. The changes of LVEDV, LVESV, and LVEF at the 1 year follow-up from the admission did not differ between the IT group and the ST group (t=0.722, P=0.444; t=1.228, P=0.221; t=1.354, P=0.187, repectively).Conclusions Short-term high-dose atorvastatin treatment for AAMI was associated with lower hs-CRP, ET-1 and MMP-9 levels compared to the standard dose treatment. However, this beneficial effect is not likely to related to the left ventricular remodeling.

Effects of Repetitive Transcranial Magnetic Stimulation Treatment on Event-Related Potentials in Schizophrenia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Repetitive transcranial magnetic stimulation (rTMS) and event-related potentials (ERPs) are a noninvasive technique that widely used in neurophysiological field. Although rTMS has shown clinical utility for a number of neurological conditions, Recently,there was little understanding of the the efficacy of rTMS on Schizophrenia(SZ) and the change of ERP between before and after rTMS treatment. The objective of this study was to investigate the characteristics of N400, mismatch negativity (MMN), and P300 before and after treatment with rTMS in SZ.</p><p><b>METHODS</b>One hundred and twenty-seven SZ patients hospitalized in Shanghai Mental Health Center from March 2015 to July 2017, divided into two groups (85 patients were recruited as rTMS group and 42 were recruited as sham rTMS [ShrTMS] group) and 76 normal controls (NCs) who were the staff and refresher staff in our hospital were recruited at the same time. A Chinese-made rTMS and a Runjie WJ-1 ERPs instrument were used in the present experiment. N400 was elicited by congruent and noncongruent Chinese idioms. After rTMS treatment, N400, P300, and MMN characteristics were compared with those before treatment and NC group.</p><p><b>RESULTS</b>Compared with NC, the SZ patients exhibited delays in N400, P300, and MMN latency and decreased N400, P300, and MMN amplitudes in their frontal area (P < 0.05). After 25 rTMS treatments, N400 amplitudes in the frontal area (elicited by idioms with same phonic and different shape and meaning and with different phonic, shape, and meaning) were increased in the SZ patients (P < 0.05). However, there was no significant change in N400 before and after treatment with ShrTMS in SZ patients (P > 0.05). Amplitudes for MMN and target P300 also increased in SZ patients after rTMS treatment (P < 0.05).</p><p><b>CONCLUSIONS</b>Based on our preliminary findings, we believe that the combined usage of N400, MMN, and P300 could be a valuable index and an electrophysiological reference in evaluating the effects of rTMS treatment in SZ patients.</p>

Construction of Lentiviral Vector Over-Expressing MEG3 and Its Effect on XG-7 Cell Apoptosis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To construct a recombinant lentiviral expression vectors carrying MEG3 and to evaluate its effects on XG-7 cell apoptosis.</p><p><b>METHODS</b>A full-length genomic fragment of human MEG3 was cloned from the pcDNA3.0-MEG3 packaging plasmid and was amplified by PCR. New restriction sites were introduced to be blunted with T4 DNA Ligase. The sequence of the amplified segments was sub-cloned into lentivirus expression vector pCDH-EF1-MCS-T2A-copGFP.The recombined lentiviral expression vector was transfected into 293T cells. FACS was used to detect the effect of MEG3 on XG-7 cell apoptosis after being infected by optimized MOI.</p><p><b>RESULTS</b>The recombined lentiviral expression vector pCDH-EF1-MEG3-copGFP was constructed successfully. The results showed that pCDH-EF1-MEG3-copGFP could increase the mRNA expression of MEG3 dramatically, its transfection efficiency was more than 90%. The apoptosis rate in XG-7 cells (26.8±2.8%) was very significantly higher than that of the control group (P<0.01).</p><p><b>CONCLUSION</b>The recombined lentiviral LncRNA expression vector targeting MEG3, pCDH-EF1-MEG3-copGFP, has been successfully constructed, the pCDH-EF1-MEG3-copGFP can induce the cell apoptosis in human myeloma cell lines. This study set up a basis to further explore the relationship between human myeloma cells and LncRNA-MEG3 gene.</p>

Follow-up of N400 in the Rehabilitation of First-episode Schizophrenia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The N400 component of event-related potentials (ERP) has recently drawn widespread attention at home and abroad. This study was to explore the relationship between N400 changes and risperidone treatment and rehabilitation infirst-episode schizophrenia (FES).</p><p><b>METHODS</b>ERP component N400 was recorded by Guangzhou Runjie WJ-1 ERP instruments, in 58 FES before and 6 months, 15 months after risperidone treatment, and in 62 normal controls. The patients' syndromes were assessed by Positive and Negative Syndrome Scale (PANSS). And the stimuli are Chinese sentences with matching (congruent) or mismatching (incongruent) ending words.</p><p><b>RESULTS</b>N400 latencies were prolonged, and amplitudes were decreased in Cz, Pz, Fz, C3, C4, in FES compared with in NC, before treatment. The prolonged N400 latencies and decreased amplitudes were negatively correlated with the patients' positive scale and total scale of PANSS. There are significant differences of N400 amplitudes and latencies in 6 months and 15 months follow-up after treatment. Before treatment, 6 months and 15 months after treatment, N400 latencies are 446 ± 35 ms, 440 ± 37 ms, 414 ± 31 ms (F = 9.72, P < 0.01) in incongruent situation; N400 amplitudes are 5.2 ± 4.6 μV, 5.7 ± 4.8 μV, 7.3 ± 5.0 μV (F = 2.06, P > 0.05) in congruent situation, and 8.5 ± 5.9 μV, 10.1 ± 5.0 μV, 11.9 ± 7.0 μV (F = 3.697, P < 0.05) in incongruent situation.</p><p><b>CONCLUSIONS</b>N400 could be used to predict the effects of treatment of schizophrenia to some degree. The linguistic and cognitive impairment in schizophrenia can be improved by antipsychotic drugs.</p>

Etiology and bone age of 2132 children with short stature / 临床儿科杂志

ObjectiveThe aim of this study is to analyze the etiology and status of bone age of children with short stat-ure.MethodsAnthropological and physical examination data were retrospectively collected and studied in 2132 children with short stature in the department of endocrinology between 2009 and 2014. Growth hormone (GH) levels were determined by ar-ginine-clonidine test. Bone age was determined by CHN scoring.ResultsAmong the 2132 patients, 1333 were males and 799 were females. Mean age is 9.03 ± 3.04 years old, mean bone age is 6.81 ± 3.05 years. Of them, 324 cases (15.2%) were diagnosed complete GH deifciency, 780 cases (36.59%) were partial GH deifciency, 27cases (1.27%) were multiple pituitary hormone de-ifciency, 13 cases (1.64%) were hypothyroidism, 893 cases (41.89%) were idiopathic short stature, 19 cases (0.89%) were small for gestational age (SGA), 40 cases (1.88%) were chromosomal disorders, etc. Signiifcant difference in age and bone age was found using t test (P<0.05). Signiifcant differences in Δage were found between etiological categories using ANOVA (P=0.000). Δage was signiifcantly and negatively associated with peak GH using Pearson's correlation.ConclusionsGH deifciency is the most common cause of short stature. Bone age of children with short stature is commonly delayed. Δage was signiifcantly and negatively associated with peak GH. Multiple pituitary hormone deifciency has a signiifcant effect on bone age. The etiology of patients with short stature cannot be determined just by bone age.

Research progress of in vivo evaluation methods for dry powder inhaler / 中国药学杂志

OBJECTIVE: To summary the common methods that used to assay dry powder inhaler in vivo, and provide a theoretical basis and some research ideas for relative research.

Correlation of zearalenone pollution with precocious puberty girls / 中华实用儿科临床杂志

Objective To explore the relationship of zearalenone (ZEA) and precocious puberty in girls.Methods The peripheral serums from 71 cases of precocious puberty girls and 50 cases of healthy girls were collected respectively and concentrations of ZEA were detected by high performance liquid chromatography.Bone age,body mass index (BMI),volume of uterus and ovaries,concentrations of estradiol (E2),luteinizing hormone (LH) and folliclestimulating hormone (FSH) were detected,and the residence of each subject was recorded as well.Results (1) In 71 patients,52 patients were diagnosed as idiopathic central precocious puberty,others were diagnosed as premature thelarche.(2) In 71 patients,serum ZEA was detected from 51 patients,and undetected in 20 patients.(3)Concentration of ZEA in precocious puberty girls [(318 ±34) ng/L]was significantly increased than that in healthy girls [(143 ± 35) ng/L,P =0.002],but no distinct difference existed between ICPP group and PT group(P =0.326).(4)Compared with uterus volume of ZEA in the undetected patients (1.975 ±0.150) cm3,the uterus volume of ZEA detected patients (2.972 ±0.180) cm3,which was significantly enlarged,there was significant difference (P =0.01) ; Percentage of overweight girls in ZEA detected patients (31.4％,16/51 cases) was lower than which in ZEA undetected patients (65.0％,13/20 cases,P =0.01) ; Although there was no statistical differences in the breast diameter,bone age,value of E2,LH and FSH between ZEA detected patients and undetected patients (all P ＞ 0.05),but the increased tendency in ZEA detected patients existed.(5) ZEA in serum was detected in 53.3％ (16/30 cases) patients living in the cities,and the rate was obviously lower than 82.9％ of the patients (34/41 cases) living in the countryside,there was significant difference (P =0.007).Conclusions ZEA is correlated with precocious puberty in girls.ZEA pollution might be one of reasons for precocious puberty occurrence.

Nonlinear dynamics of electroencephalography study in schizophrenic patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Few characteristic changes of linear electroencephalograph (EEG) have been reported in schizophrenia. The aim of the present study was to investigate the changes in temporal-spatial dimensional properties of EEG under different cognitive tasks in patients with schizophrenia.</p><p><b>METHODS</b>EEG was recorded by using EEG-1518K system and mapping system (Nihon Kohden Tomioka Corporation, Japan) in 45 schizophrenic patients and 47 healthy adults (normal control, NC) under five states: eyes closed, eyes open, mental arithmetic test with eyes closed, memory test with eyes open, and number cancellation test. Correlation dimension (D2) and point-wise correlation dimension (PD2) were calculated for all EEG analyses.</p><p><b>RESULTS</b>(1) There were no significant differences of D2 and PD2 between NC and schizophrenic patients under states of eyes open and closed. (2) Compared with NC, schizophrenic patients showed decreased performance of D2 in mental arithmetic test with eyes closed and number cancellation test (mental arithmetic test with eyes closed: Nc 5.9 ± 0.6, Sch 3.0 ± 0.8; number cancellation test: Nc 6.0 ± 0.6, Sch 4.4 ± 0.7; P < 0.05 or P < 0.01). (3) Schizophrenic patients also showed decrease performance of PD2 in mental arithmetic test with eyes closed, memory test with eyes open, and number cancellation test (mental arithmetic test with eyes closed: Nc 6.9 ± 0.7, Sch 4.0 ± 0.8; memory test with eyes open: Nc 6.6 ± 0.8, Sch 5.0 ± 0.9; number cancellation test: Nc 7.1 ± 0.7, Sch 4.8 ± 0.9; P < 0.05 or P < 0.01).</p><p><b>CONCLUSIONS</b>Nonlinear dynamic analysis provided a new approach in clinical investigation of EEG signals. It was helpful to further understand the cerebral mechanism in schizophrenic cognitive process.</p>

Neonatal ventral hippocampal lesion as a valid model of schizophrenia: evidence from sensory gating study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The neonatal ventral hippocampal lesion (NVHL) rat model has been proposed as an experimental model for schizophrenia. NVHL rats display impaired central nervous system (CNS) inhibition, which may lead to a phenomenon similar to P50 sensory gating deficits observed in schizophrenic patients. In this study, we investigated whether sensory gating deficits occurred in the NVHL rat as a model for schizophrenia.</p><p><b>METHODS</b>We created the NVHL rat model using ibotenate. The P20 and N40 were measured to assess sensory response and gating in NVHL and sham rats. Epidural electrodes recorded evoked potentials (EPs), from which latencies, amplitudes, difference scores (S1-S2), and gating ratios (S2/S1) were assessed.</p><p><b>RESULTS</b>Compared with sham controls, prolonged S1 N40 latency and decreased S2 N40 amplitude were detected in the NVHL group. In neither difference scores nor gating ratios, a significant difference was found between NVHL group and sham controls.</p><p><b>CONCLUSIONS</b>NVHL rats may be a valid animal model for schizophrenia. This strategy will be useful in future neurobiological studies investigating the etiology of schizophrenia.</p>

The attenuating effect of ursodeoxycholic acid on endoplasmic reticulum stress-mediated pancreatic β-cell apoptosis in streptozotocin-induced diabetic rats / 中华内分泌代谢杂志

Objective To clarify the protective effect of nrsodeoxycholic acid ( UDCA ) on endoplasmic reticulum stress-mediated apoptosis in pancreatic β-cell of streptozotocin ( STZ )-induced diabetic rats.Methods Rats( n =40) received a single injection STZ( 50 mg/kg) intra-peritoneally and formed a β-cell injury model.Weight-matched normal rats( the control group,n =10 ) were injected with the buffer alone.STZ-treated rats with persistent random blood glucose higher than 16.7 mmol/L for 1 week were considered as diabetic status( n=14 ),then divided randomly into STZ-induced diabetes mellitus ( DM ) group ( n =7 ) and UDCA-treated DM group ( n =7 ).UDCA (40 mg· kg- 1,d-1 ) was administered daily by intragastric intubations throughout the experimental period (30 d).During the entire experiment,blood glucose in all rats was assessed.By the end of the experiment,all rats were sacrificed with the pancreas removed and the blood sample collected immediately.Fasting insulin levels were assayed by radioimmunoassay.The morphological changes of pancreatic β-cells apoptosis were determined by TUNEL assay.RNA in pancreas was abstracted and microarray containing 89 pieces of apoptosis related genes was applied.The related gene expressions were detected by RT-PCR and Western blot.Results The concentration of blood glucose in diabetic rats was gradually decreased after UDCA treatment,but at the end of the experiment it was still higher than that in the normal control group.The treatment with UDCA raised the fasting insulin level in diabetic rats,but this concentration was significantly lower as compared to the control group.Based on TUNEL stained tissue sections,the percentage of β-cell apoptosis of UDCA-treated DM group was significantly lower than that of STZ-induced diabetic group(P＜0.05 ).Among 89 genes,42 genes up-regulated and 46 genes down-regulated in diabetic rats,some of which were ameliorated by UDCA treatment.The expressions of Caspase-3,Bax,Bip,and CHOP mRNA in pancreas of DM group were significantly up-regulated as compared with those in the control group ( P ＜ 0.05 ) ; while the expression of Bcl-2 mRNA was markedly down-regulated (P＜0.05 ).However,these parameters in the U DCA-treated animals showed a marked improvement.Conclusion Ursodeoxycholic acid seems to protect pancreatic β-cell from apoptosis in STZ-induced diabetes by attenuating the severity of endoplasmic reticulum stress.

Influence of trichostatin A on apoptosis of colon carcinoma cells and expression of HIF-1α under hypoxia in vitro / 第二军医大学学报

Objective: To observe the influence of trichostatin A (TSA) on the apoptosis of colonic cancer cell line HT-29 cells and expression of HIF-1α in HT-29 cells under hypoxia in vitro. Methods: HT-29 cells were treated with TSA(100, 200, 400, 600, 800 nmol/ L for 12, 24, 48, and 72 h. Cells in the control group were treated with the same volume of DMSO. MTT assay was used to investigate the survival of cells in each group. Annexin V and TUNEL were used to determine the apoptosis rate of HT-29 cells. RT-PCR and Western blotting were used to study the expression of HIF-1α mRNA and protein under hypoxia (37°C, 1% O2). Results: MTT assay demonstrated that cell viability decreased significantly in TSA-treated group compared to control group(P<0.05), and the inhibitory effect of TSA was in a dose- and time-dependent manner. Annexin V and TUNEL showed that TSA induced early apoptosis of HI-29 cells; the apoptosis rates in TSA treatment groups (200, 400, 600, and 800 nmol/L) were significantly higher than that of control group(P<0.05). TSA suppressed protein expression of HIF-1α under hypoxia in a dose-dependent manner but had no influence on mRNA expression of HIF-1α. Conclusion: TSA can induce apoptosis of HI-29 cells and inhibit their proliferation through suppressing expression of HIF-1α at protein level under hypoxia condition.

Application of cook MOB-15 system in guiding wire insertion during endoscopic retrograde cholangiopancreatography / 第二军医大学学报

Objective: To evaluate the value of Cook MOB-15 system in guiding wire insertion during endoscopic retrograde cholangtopancreatography (ERCP). Methods: The clinical data of 51 patients who received Cook MOB-15 system-guided wire insertion during ERCP between Jan. 2005 to Dec. 2007 were retrospectively analyzed. Forty patients who received conventional ERCP catheter for malignant jaundice between Jan. 2002 and Dec. 2004 were taken as control. The successful insertion rates were compared between the 2 groups. Results: The successful insertion rate was 90.2% (46/51) in the Cook MOB-15 system group and 72.5% (29/40) in the conventional group; there was significant difference between the 2 groups (P<0. 05). Conclusion: The Cook MOB-15 system can significantly raise the successful insertion rate in ERCP.

Different strategies for preparation of non-tagged rV270 protein and its efficacy against Yersinia pestis challenge / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>LcrV is an important component for the development of a subunit vaccine against plague. To reduce immunosuppressive activity of LcrV, a recombinant LcrV variant lacking amino acids 271 to 326 (rV270) was prepared by different methods in this study.</p><p><b>METHODS</b>A new strategy that produced non-tagged or authentic rV270 protein was designed by insertion of rV270-thrombin-hexahistidine fusion gene into the vector pET24a, or by insertion of hexahistidine-enterokinase-rV270 or hexahistitine-factor Xa-rV270 fusion gene into the vector pET32a. After Co(2+) affinity chromatography, a purification strategy was developed by cleavage of His tag on column, following Sephacryl S-200HR column filtration chromatography.</p><p><b>RESULTS</b>Removal of His tag by thrombin, enterokinase and factor Xa displayed a yield of 99.5%, 32.4% and 15.3%, respectively. Following Sephacryl S-200HR column filtration chromatography, above 97% purity of rV270 protein was obtained. Purified rV270 that was adsorbed to 25% (v/v) Al(OH)₃ adjuvant in phosphate-buffered saline (PBS) induced very high titers of antibody to rV270 in BALB/c mice and protected them (100% survival) against subcutaneous challenge with 10⁶ CFU of Y. pestis virulent strain 141.</p><p><b>CONCLUSION</b>The completely authentic rV270 protein can be prepared by using enterokinase or factor Xa, but they exhibited extremely low cleavage activity to the corresponding recognition site. Thrombin cleavage is an efficient strategy to prepare non-tagged rV270 protein and can be easily operated in a large scale due to its relatively low cost and high cleavage efficacy. The recombinant rV270 can be used as a key component to develop a subunit vaccine of plague.</p>

N400 elicited by incongruent ending words of Chinese idioms in healthy adults / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Prior research about N400 has been mainly based on English stimuli, while the cognitive processing of Chinese characters is still unclear. The aim of the present study was to further investigate the semantic processing of Chinese idioms.</p><p><b>METHODS</b>Event related potentials (ERP) component N400 was elicited by 38 pairs of matching (congruent) and mismatching (incongruent) ended Chinese idioms: ending words with same phoneme but different shape and meaning (sPdSdM), with similar shape but different phoneme and meaning (sSdPdM), with same meaning but different phoneme and shape (sMdPdS), and words with different phoneme, shape and meaning (dPdSdM) and recorded by Guangzhou Runjie WJ-1 ERP instruments. In 62 right-handed healthy adults (age 19 - 50 years), N400 amplitudes and latencies were compared between matching and mismatching conditions at Fz, Cz and Pz.</p><p><b>RESULTS</b>N400 showed a midline distribution and could be elicited in electrodes Fz, Cz and Pz. The mean values of N400 latencies and amplitudes were obtained for matching and mismatching ending words in healthy adults. Significant differences were found in N400 latencies and amplitudes in matching and mismatching ending-words idioms in healthy adults (P < 0.05). Compared with matching ending-words idioms, N400 latencies were prolonged and the amplitudes were increased in mismatching ones. N400s elicited by different types of stimuli showed different latencies and amplitudes, and longest N400 latency and largest N400 amplitude were elicited by ending-words with dPdSdM. No gender difference was found of N400 latency and amplitude in this study (P > 0.05).</p><p><b>CONCLUSIONS</b>Compared with English stimuli, Chinese ideographic words could provide more flexible stimuli for N400 research in that the words have 3-dimension changes - phoneme, shape and meaning. Features of N400 elicited by matching and mismatching ending words in Chinese idioms are mainly determined by the meaning of the word. Some issues of N400 elicited by Chinese characters deserve further research.</p>

Color Doppler ultrasonic diagnosis of acute osteofascial compartment syndrome of forearm / 中国介入影像与治疗学

Objective To assess the application value of color Doppler ultrasound (CDU) in the diagnosis of osteofascial copmarttnent syndrome (OCS) of forearm.Methods A total of 19 patients highly suspected of or diagnosed as acute OCS of forearm underwent CDU in comparison with clinical diagnosis and therapy.The echoes of muscles,the diameter and change of flow Doppler spectra of radial artery and ulnar artery were observed.Results In 16 patients of OCS,most of the forearm muscle swelled (enlarged more than 1/2-2/3 area) with the muscle echo enhanced unevenly:the textures were unclear or disappeared;and fluid or hematom appeared in muscles;and the diameter of radial artery and ulnar artery diminished.The flow Doppler spectra of radial artery and ulnar artery appeared as bid-diastolic whole inverse wave,bip-,unid-,uniph-wave,unip-,unid-uniph-wave and venous like Doppler wave.Resistance index increased.Conclusion The acute OCS of forearm has some sonographic characteristics.CDU can be used to diagnose the acute OCS of forearm.

Determination of cotinine, phenylglyoxylic acid and mandelic acid in human urine by GC/MS / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a GC/MS method for analysis of cotinine (COT), phenylglyoxylic acid (PA) and mandelic acid (MA) in human urine.</p><p><b>METHODS</b>Human urine samples were extracted by CCl(3) and derivatized with MSTFA after dried completely. The contents of COT, PA and MA were measured by GC/MS method with DB-5MS capillary column and EI ion-source.</p><p><b>RESULT</b>The calibration curves for COT in urine samples were linear over the concentration ranges of 0.0002 approximately 3.5 microg ml(-1), while PA and MA were both of 1.25 approximately 160 microg ml(-1). The limits of quantification were 0.0002 microg ml(-1), 1.25 microg ml(-1) and 1.25 microg ml(-1) for COT, PA and MA, respectively. The assay recoveries for COT, PA and MA ranged from 89.53% approximately 102.4%, 84.88% approximately 91.46% and 83.46% approximately 13.6%, respectively.</p><p><b>CONCLUSION</b>The established method can detect cotinine, phenylglyoxylic acid and mandelic acid simultaneously, which would be used in routine assessment and monitoring of the internal exposure to nicotine and styrene in human body.</p>