The Regulation of Reactive Oxygen Species on Dendritic Cells / 中国生物化学与分子生物学报

Dendritic cells (DCs) are specialized antigen-presenting cells. Immature dendritic cells can be activated into mature dendritic cells by recognizing antigens, then the antigens are processed and pres-ented to T lymphocytes. DCs play a vital role in initiating immune response, regulating immune response and maintaining immune tolerance. Therefore, regulating the immune function of DCs can be used to treat diseases such as autoimmune diseases and tumors. With the deepening of research on the regulation of DCs, people have gradually realized that the existence of reactive oxygen species (ROS) in DCs is of great significance. ROS is a term of strong oxidizing reactive species, and the dynamic balance of its pro-duction and removal is the key to maintaining cell redox homeostasis. ROS of physiological level is an im-portant molecule involved in a variety of signal pathways, which can regulate cell growth, differentiation and different physiological and biochemical reactions. The changes in the level of ROS affect the state and function of cells. In addition, because there are many ways to produce ROS in the cell, the effects of dif-ferent sources of ROS on DCs are not usually the same; and even the same source of ROS may have dif-ferent effects when cells are in different states. This article summarized the influence of intracellular ROS changes and different sources of ROS on the differentiation, maturation and function of DCs, aiming to re-veal how ROS plays an important role in regulating the immune function of DCs. At the same time, this article also showed the urgent need for the in-depth study of ROS regulating the function of DCs, which may help the application of DCs immune regulation to a wider range of disease treatments.

Hypoxic microenvironment promotes the proliferation of human olfactory mucosa mesenchymal stem cells and its associated mechanism / 中国组织工程研究

BACKGROUND: Human olfactory mucosa mesenchymal stem cells (hOM-MSCs) not only have the basic characteristics of mesenchymal stem cells, but also originate from the ectoderm and are prone to differentiate into neurons, which are a kind of ideal seed cells for nerve repair and regeneration. Cells are conventionally cultured in about 21% in vitro, while only 3%-9% oxygen is found in the human body and tissue space. There is still no report on the effect of hypoxia on the proliferation and activity of hOM-MSCs.OBJECTIVE: To explore whether hypoxic microenvironment can promote hOM-MSCs proliferation and activity and the related mechanism. METHODS: hOM-MSCs were isolated, cultured and identified by flow cytometry and immunofluorescence. The passage 4 hOM-MSCs were divided into three groups: 21% O2 group, 3% O2 group and 3% O2+20 μmol/L YC-1 (HIF-1α inhibitors) group. Proliferation and apoptosis of hOM-MSCs was detected by flow cytometry after 72 hours of culture. The proliferating cell nuclear antigen protein expression was detected by western blot. The mRNA and protein expression of HIF-1α and TWIST were detected by Q-PCR and western blot. RESULTS AND CONCLUSION: The purity of hOM-MSCs was up to 97%, as defined by flow cytometry. The proliferation index of 3% O2 group was higher than the 21% O2 group (P < 0.05), and cell survival and apoptosis ratio (apoptotic cells included mechanical death + early apoptosis + late apoptosis) between the two groups had no significant difference (P > 0.05). Western blot results showed that the proliferating cell nuclear antigen protein expression in the 3% O2 group was significantly higher than that in the other groups (P < 0.05). The HIF-1α and TWIST expressions at mRNA and protein levels in the 3% O2 group were significantly higher than those in the other groups (P < 0.05). To conclude, hypoxic microenvironment can promote the hOM-MSCs proliferation and has no effect on the apoptosis, and the HIF-TWIST signal pathway plays an important role in this progress.

Preliminary investigation of the effect of CO2 laser assisted stapedotomy on the treatment of advanced otosclerosis / 中国耳鼻咽喉头颈外科

OBJECTIVE To explore the outcomes of CO2 laser assisted stapedotomy with artificial stapes prostheses in the treatment of advanced otosclerosis. METHODS Between January 2010 and January 2014, 15 patients (16 ears) diagnosed as advanced otosclerosis accepted CO2 laser assisted stapedotomy with artificial stapes implantation in our department. The averaged preoperative air conduction threshold of the speech frequency was 70.21 dB HL, the averaged bone conduction threshold was 38.49 dB HL, the averaged air-bone gap (ABG) was 31.72 dB HL. All cases were followed up for more than 6 months after operation. RESULTS All cases accepted auditory follow up after 6 months postoperatively. The speech frequency average air conduction threshold was 43.7 dB HL, the average bone conduction threshold was 28.95 dB HL, the average ABG was 14.75 dB HL. The ABG≤20 dB was achieved in 9 ears (56.3%) and ABG closure (≤10 dB) was achieved in 6 ears (37.5%). No cases appeared intractable vertigo, sensorineural hearing loss, secondary facial paralysis and other serious complications. CONCLUSION CO2 laser assisted technique reduced the probability of serious complications of stapedotomy, most patients with hearing level improved significantly. It's a safe, practical, relatively economical choice for advanced otosclerosis.

Discussion on twirling reinforcing-reducing method / 中国针灸

The essence of twirling reinforcing-reducing method is discussed to provide theoretical guidance for clinical application of reinforcing-reducing method. Through retrospection on historical literature of twirling reinforcing-reducing method, records and explanatory notes are thoroughly explored. Several existing opinions are analyzed and explained for instance twirling method has connection with circulation direction of channels; twirling method is subdivided into right and left, male and female, hand and foot; twriling method is related to quantity of stimulus and operation time; twriling method belongs to spiral motion and so on. As a result, it is found that the key of twirling reinforcing-reducing method is the posture of needle-holding hand that defines three-dimensional motion. If twirling method is subdivided into right and left, male and female, hand and foot and so on, steric effects of lifting-thrusting movement that come along with twirling method could be ignored at the same time. It is that the essence of twirling reinforcing-reducing method is close to the principle of lifting-thrusting reinforcing-reducing method, enriching effect with slow insertion and fast withdrawal of needle while reducing effect with fast insertion and slow withdrawal, which is recorded in Miraculous Pivot: Nine needle and Twelve Yuan. With this principle as guide, manipulation could be avoided to become a mere formality and illusory metaphysics during clinical application of twirling reinforcing-reducing method.

Effects of Yindan Xinnaotong Capsule on OPN expression in myocardium tissue of rats after acute myocardial infarction and its prevention mechanism on myocardium / 吉林大学学报(医学版)

Objective To investigate the effects of Yindan Xinnaotong Capsule on the osteoponin(OPN)expression in myocardium tissue of the rats after acute myocardial infarction(AMI)and to clarify the mechanism of Yindan Xinnaotong Capsule in alienating rat AMI.Methods 90 Wistar rats were used to establish AMI models by ligating of the left anterior descending coronary artery.The AMI model rats were randomly divided into AMI model group, highdoseofYindan(16g·kg-1·d-1)group,lowdoseofYindan(0.08g·kg-1·d-1)group,positivedrug control captopril(5 mg·kg-1 ·d-1 )group(n=12);at the same time sham operation group(n=10)was set up, the rats in sham operation group was treated with wearing without ligation. All the rats were administrated for 4 weeks,then the myocardium tissue samples were obtained.The histological changes of myocardium tissue were observed by HE and Masson staining;the DNA fragments of apoptotic cells were detected by TUNEL staining and the apoptotic index(AI)was calculated.The expression of OPN mRNA in non-infarction area was measured by RT-PCR.Results Compared with sham operation group,the AI of the rats in model group was significantly increased (P<0.05);compared with model group,the AI of the rats in Yindan groups and captopril group were markedly decreased(P<0.01 );compared with captopril group, the AI of the rats in high dose of Yindan group was significantly decreased(P<0.05).Compared with sham operation group,the expression level of OPN mRNA in non-infarction area of the rats in model group was significantly increased(P<0.01);compared with model group, the expression levels of OPN mRNA in non-infarction area of the rats in Yindan groups and captopril group were also decreased significantly(P<0.01);compared with captopril group,the expression level of OPN mRNA non-infarction area of the rats in high dose of Yindan group was significantly decreased(P<0.05).Conclusion Yindan Xinnaotong Capsule could obviously alleviate the apoptosis of myocardium tissue of the rats after acute myocardial infarction and decrease the expression of OPN mRNA in non-infarction area of left ventricle, which indicates that Yindan Xinnaotong Capsule may protect myocardium tissue through decreasing the OPN mRNA expression.

Two strategies to intensify evidence-based medicine education of undergraduate students: a randomised controlled trial

<p><b>INTRODUCTION</b>Undergraduate evidence-based practice (EBP) is usually taught through standalone courses and workshops away from clinical practice. This study compared the effects of 2 clinically integrated educational strategies on final year medical students.</p><p><b>MATERIALS AND METHODS</b>Final year medical students rotating to the general medicine service for a 2-week internship were randomly assigned to participate in a weekly EBP-structured case conference focusing on students' primary care patients (Group A, n = 47), or to receive a weekly didactic lecture about EBP (Group B, n = 47). The teaching effects of these 2 interventions were evaluated by a validated instrument for assessment of EBP related knowledge (EBP-K), attitude (EBP-A), personal application (EBP-P), and anticipated future use (EBP-F) on the first and last days of rotation.</p><p><b>RESULTS</b>All scores improved significantly after the 2-week EBM-teaching for both groups. When compared to Group B, students in Group A had significantly higher post-intervention scores of EBP-K (21.2 ± 3.5 vs 19.0 ± 4.6; ie. 57.8 ± 72.9% vs 29.1 ± 39.1%; P <0.01) and EBP-P (18.7 ± 4.3 vs 15.3 ± 3.9; ie. 28.5 ± 25.5 % vs 14.1 ± 18.7 %; P <0.001). In contrast, the scores of EBP-A and EBP-F were similar between the 2 groups.</p><p><b>CONCLUSION</b>Structured case conference, when compared to the didactic lectures, significantly improved EBP-K and EBP-P for final year medical students.</p>

Professor DONG Gui-rong's experience for the treatment of peripheral facial paralysis / 中国针灸

Professor DONG Gui-rong's theoretical principle and manipulation points for peripheral facial paralysis were introduced in details from the angels of syndrome differentiation, timing, acupoint prescription and needling methods. For the syndrome differentiation and timing, the professor emphasized to check the treatment timing and follow the symptoms, which should be treated by stages, besides, it was necessary to find and distinguish the reason and nature of diseases to have a combined treatment of tendons and muscles. For the acupoint prescription and needling methods, he has proposed that the acupoints selection should be compatible of distal and lacal, and made a best of Baihui (GV 20) to regulate the whole yang qi, also he has paid much attention to the needling methods and staging treatment. Under the consideration of late stage of peripheral facial paralysis, based on syndrome differentiation Back-shu points have been selected to regulate zang-fu function, should achieve much better therapeutic effect.

Clinical observation on the principle of "single usage of acupoints of Shaoyang meridian" for treatment of facial paralysis in acute stage / 中国针灸

<p><b>OBJECTIVE</b>To compare different therapeutic effects between single usage of acupoints of Shaoyang meridian and the routine ones for treatment of Bell palsy in acute stage.</p><p><b>METHODS</b>One hundred and twenty cases with Bell palsy during the first three days were random divided into an observation group and a control group, 60 cases in each group. In observation group, acupoints of Shaoyang meridians were used from the 3rd day till the 14th day, Fengchi (GB 20), Yifeng (TE 17), Wangu (GB 12) at the affected side etc. were selected, after the 15th day, the routine acupoints were applied, Hegu (LI 4) on both sides, Fengchi (GB 20), Quchi (LI 11), Yangbai (GB 14) at the affected side etc. were selected; the control group were treated with the same acupoints as the routine ones in observation group since the 3rd day. And both two groups were treated with oral administration of Prednisone. House-Brackmann (H-B) functional grading of facial nerve on the 3rd day with the one of the 60th day as well as electroneurography (ENoG) on the 3rd day with the one of the 14th day were compared respectively.</p><p><b>RESULTS</b>The H-B grading improvement and cured rate were 95.0% (57/60) in observation group, which were suprior to 83.3% (50/60) in control group; the cured time in observation group was (34.21 +/- 8.026) days, significantly shorter than (42.78 +/- 9.029) days in control group (P < 0.05).</p><p><b>CONCLUSION</b>On the basis of oral administration of Prednisone, single usage of acupoints of Shaoyang meridian in acute stage can make great improvement for recovery of Bell palsy, better than routine point selection.</p>

Side effects of L-asparaginase during therapies for remission induction and maintenance in children with acute lymphocytic leukemia / 中国实验血液学杂志

This study was purposed to investigate the possible side effects of L-asparaginase (L-ASP) in the treatment of patients with acute lymphoblastic leukemia (ALL) and to explore the correlation of these side effects at different therapeutic stages by means of retrospective analysis, so as to reduce the incidence of side effects and improve the safety of chemotherapy and the long-term survival of patients. The probability and severity of side effects related to use of L-ASP in 38 cases of ALL during remission induction therapy (VDLDex regimen) and 40 cases of ALL during maintenance intensive therapy (VMLDex regimen) were compared. The results showed that allergic response, diabetes and drug-induced liver disease happened more frequently during maintenance therapy than during remission induction therapy, while defibrination, abnormal hemagglutinin, acute pancreatitis, hypoproteinemia, gastrointestinal reaction and infectious shock were observed more during remission induction therapy than those at maintenance therapy. In conclusion, the L-ASP showed some side effects especially for the patients during the remission induction therapy which should be paid enough attention. The regular and comprehensive monitoring can effectively reduce and avoid the side effects of L-ASP, to improve the safety of chemotherapy.

Clinical study on acupuncture intervention time for treatment of peripheral facial paralysis / 中国针灸

<p><b>OBJECTIVE</b>To observe effects of intervention time of local acupuncture at the affected side on the facial nerve injury and the therapeutic effect at acute stage of peripheral facial paralysis.</p><p><b>METHODS</b>Two hundred and seventy-nine cases within 3 days of attack were randomly divided into 4 groups, group A (n=74), group B (n=70), group C (n=74) and control group (n=61). The 4 groups were treated with Prednisone on the third day after attack, and acupuncture was added in the group A, B and C, with Fengchi (GB 20), Yangbai (GB 14), Taiyang (EX-HN 5), Sibai (ST 2), Yingxiang (LI 20), etc. on the affected side and bilateral Hegu (LI 4) selected, and with superficial insertion method used for acupoints on the ear-face parts without manipulating the needles, and electroacupuncture was added from the fifth session of the treatment, and uniform reinforcing-reducing method was used for the distal acupoints selected. The needles were retained for 20 min and the treatment was given for 25 sessions, once other day. The therapeutic effects, the mean therapeutic courses for the cured patients and changes of electroneurography (ENoG) were compared among the groups.</p><p><b>RESULTS</b>The clinical total effective rate was 98.6%, 95.7%, 94.6% and 72.1% in the group A, B, C and the control group, respectively, with a significant difference (P < 0.05), and the therapeutic course for the cured patients increased in the order of the group A, B, C and the control group; and there was no significant difference among the 4 groups in changes of ENoG at the third day and the fourteenth day (both P > 0.05).</p><p><b>CONCLUSION</b>Acute stage is the best opportunity for acupuncture treatment of peripheral facial paralysis, and the earlier the intervention time, the better the therapeutic effect and the shorter the therapeutic course.</p>

Postnatal and prenatal diagnosis of mucopolysaccharidosis type III (Sanfilippo syndrome) / 中华儿科杂志

<p><b>OBJECTIVE</b>Mucopolysaccharidosis (MPS) types IIIA, B, C, D are a group of autosomal recessive lysosomal storage disorders caused by mutations in one of four genes which encode enzyme activities required for the lysosomal degradation of heparan sulfate. MPSIIIA and MPSIIIB involve deficiencies of heparan N-sulfatase (SGSH) and alpha-N-acetylglucosaminidase (NAGLU). MPS IIIA and MPS IIIB are more common than MPS IIIC and IIID. The present study aimed to establish two enzyme assay methods for SGSH and NAGLU activities for carrying out postnatal and prenatal diagnosis of MPSIIIA and IIIB by means of SGSH and NAGLU activity assay on plasma, leukocyte, uncultured chorionic villi (CV) and cultured amniotic fluid cells (AF cell) using two newly synthesized substrates. Mutation analysis of SGSH gene was also performed.</p><p><b>METHODS</b>Two fluorigenic substrate (4-methylumbelliferyl-alpha-D-N-sulphoglucosaminide.Na and 4-methylumbelliferyl-alpha-N-acetylglucosaminide) were used for the assay of SGSH and NAGLU activity. SGSH activity in leukocyte was determined for diagnosis MPSIIIA proband. NAGLU activity was determined in plasma for diagnosis of MPSIIIB proband. Twelve cases with MPS III were enrolled in this study, 4 were female and 8 were male, age 3 - 10 years and were from 10 unrelated families. Eight exons of SGSH gene were amplified by PCR. The mutations of the patients were characterized by direct sequencing of the amplified DNA fragments. Prenatal diagnosis in 3 pregnancies at risk was carried out according to NAGLU activity on uncultured CV at 11th week or on cultured AF cell at 18th week of gestation.</p><p><b>RESULTS</b>The SGSH activities in leukocyte of normal controls were 4.4 - 8.1 nmol/(17 h.mg protein). The NAGLU activity in plasma of normal controls was 33.3 - 62.4 nmol/(4 h.ml). The NAGLU activities were 44.9 - 91.7 nmol/(17 h.mg protein) and 53.2 - 82.2 nmol/(17 h.mg protein) in CV and cultured AF cells respectively. Five cases of MPS IIIB and 7 cases of MPS IIIA were diagnosed. The mutation analysis of SGSH gene showed 6 mutations (G191R, D235N, R377C, E447K, R233X and D219Wfs264X), only one of which (D219Wfs264X) has not been previously reported. Prenatal diagnosis was performed on 3 pregnancies at risk. NAGLU activity of one affected fetus was 1.5 nmol/(17 h.mg protein) in AF cell.</p><p><b>CONCLUSIONS</b>The method using synthesized fluorigenic 4-methylumbelliferyl-substrates were sensitive, rapid and convenient assay of SGSH and NAGLU activity and were reliable for early prenatal diagnosis. Mutation analysis on MPS IIIA patients suggests new possibilities for molecular diagnosis of the disease.</p>

Purification and Characterization of a Recombinant Thermostable ?-mannanase from Thermotoga maritima / 微生物学通报

A ?-mannanase gene (TM_1227) from Thermotoga maritima MSB8 was cloned and expressed in E.coli. The recombinant ?-mannanase was purified and characterized. The gene consists of 2010 bp, and the translated protein encodes 669 amino acids and its molecular mass is approximately 76.827 kD. Homology analysis of the deduced amino acid sequences showed that the enzyme shared 99% identity with ?-mannanase from Thermotoga sp. RQ2. The mannanase activity was up to 39.7 U/mg after the recombinant E. coli BL21 was induced by IPTG. Crude enzyme solution was purified to homogeneity by Ni-NTA agarose. Its optimum temperature and pH was 95?C and pH 8.0 respectively for LBG. The enzyme remained over 50% activity after treated at 85?C for 30 min. The above properties showed great potential of its application in paper industry. The mannanase hydrolyzed copra mannan and LBG to give various sizes of oligosaccharides, and almost no mannose was detected by TLC, which was suitable for mannooligosaccharides production.

Postnatal and prenatal diagnosis of mucopolysaccharidosis type II (Hunter syndrome) / 中华儿科杂志

<p><b>OBJECTIVE</b>Mucopolysaccharidosis type II (MPS II, Hunter syndrome, OMIM 309900) is an X-linked recessive lysosomal storage disease resulting from a deficiency of iduronte-2-sulphate sulphatase (IDS). The present study aimed to establish an enzyme assay method for IDS activity for carrying out postnatal and prenatal diagnosis of MPS II by means of IDS activity assay on plasma, uncultured chorionic villi (CV) and cultured amniotic fluid cells (AF cell) using a new synthesized substrate.</p><p><b>METHODS</b>A fluorigenic substrate (4-methylumbelliferyl-alpha-iduronate-2-sulphate, MU-alpha-Idu-2S) was used for the assay of IDS activity. IDS activity in plasma was determined for diagnosis of the proband. Prenatal diagnosis in 10 pregnancies at risk was carried out according to IDS activity on uncultured CV at 11th week or on cultured AF cell at 18th week of gestation. At the same time, IDS activity was also determined in the maternal plasmas to observe the change of IDS activity in pregnancy. The fetal sex determination was performed by PCR amplification of the ZFX/ZFY genes.</p><p><b>RESULT</b>The IDS activity in plasma of normal controls and obligate heterozygotes were 240.2 - 668.2 nmol/(4 hxml) and 88.7 - 547.9 nmol/(4 hxml), respectively, while the enzyme activities in plasmas were in the range of 0.3 - 18.6 nmol/(4 hxml) in affected male. The IDS activities were 37.2 - 54.9 nmol/(4 hxmg protein) and 21.4 - 74.4 nmol/(4 hxmg protein) in CV and cultured AF cells respectively. Out of 50 suspected cases, 46 were diagnosed as having MPS II and 4 were excluded. Prenatal diagnosis was performed on 10 pregnancies at risk. Four of 5 male fetuses [IDS activity were 4.7, 1.8, 7.0 nmol/(4hxmg protein) in CV, 0.6 nmol/(4 hxmg protein) in AF cell] were diagnosed as having MPS II and the other 5 fetuses were normal females [IDS activity were: 48.7, 5.9, 25.2 nmol/(4 hxmg protein) in CV, 55.2, 40.9 nmol/(4 hxmg protein) in AF cell]. Increased IDS activity was observed in plasma of the pregnant women with unaffected fetuses, while the IDS activity decreased in pregnancies with affected fetuses. IDS activity of one female fetus was very low [5.9 nmol/(4 hxmg protein)], but the IDS activity in maternal plasmas increased, this fetus was a normal female.</p><p><b>CONCLUSIONS</b>The method using a synthesized fluorigenic 4-methylumbelliferyl-substrate was a sensitive, rapid and convenient assay of IDS activity and was reliable for early prenatal diagnosis. Determination of fetal sex would be helpful in excluding the female fetus with low IDS activity from being considered as an affected male fetus. It would be further helpful if IDS activity in maternal plasma was taken into account.</p>

Optimization of Xylanase Production by Paecilomyces thermophila in Solid State Fermentation / 微生物学通报

A new thermophilic fungus J18 isolated from the soil samples was identified as Paecilomyces thermophila. This strain produced effectively xylanase utilizing several lignocellulosic materials in the solid-state fermentation (SSF) , and wheat straw was the best carbon source. The results of single-factor-experiment showed that the wheat straw of particle size 0. 3 mm ~ 0.45 mm, initial moisture content of 83% , initial pH of 7. 0 and cultivation temperature of 50℃were the optimal conditions for xylanase production. Under the optimized conditions, it produced 18 580 U/g dry substrate after 8 days of cultivation. Therefore, xylanase production by Paecilomyces thermophila in SSF possess great potential for commercial applications.

IDENTIFICATION OF A XYLANASE PRODUCING STRAIN OF STREPTOMYCES SP. AND OPTIMIZATION OF CONDITIONS ON ITS ENZYME PRODUCTION / 微生物学通报

A xylanase producing strain was screened with xylan as the only carbon source. The strain was identified as Streptomyces cirratus. The effects of different factore on the enzyme production were studied. Corncobs xylan (water insoluble) and tryptone were the best C and N sources, respectively. The enzyme activity was increased to about 2.5 times by addition of 0.5% Tween 80 in the medium. The highest xylanase activity was up to 623u/mL.

Cloning and Expression of a?-galactosidase Gene from Thermotoga maritima / 微生物学通报

The cloning and expression of a?-galactosidase gene(TM_0310)from Thermotoga maritima MSB8 was studied.The gene consists of 2019 bp,and the translated protein encodes 672 amino acids and its molecular mass is approximately 78.972 kD.The homology analysis of the deduced amino acid sequences showed that the enzyme shared 95%identity with a putative?-galactosidase from Thermotoga petrophila RKU-1 and a?-galactosidase from Thermotoga sp.RQ2.The galactosidase activity was up to 2.08 U/mg after the recombinant E.coli BL21 was induced by IPTG.The crude lysate remained about 70%activity after treated at 80℃for 10 min,indicating that the recombinant enzyme is thermostable and may be used at high temperatures.