Breaking blood expelling stasis method accelerates hematoma resolution after hyperacute intracerebral hemorrhage and its potential mechanism / 中国医师杂志

Objective:To explore the efficacy of breaking blood expelling stasis method accelerates hematoma resolution after intracerebral hemorrhage (ICH) and its potential mechanism.Methods:63 ICH patients confirmed by computer tomography (CT) scan from August 2019 to February 2020 were selected as the research objects and randomly divided into control group ( n=29, routine treatment plus placebo) and observation group ( n=34, routine treatment plus breaking blood expelling stasis granules). The changes of neurological function and hematoma volume were observed before and after treatment. At the same time, the ICH rat model was constructed to observe the changes of neurobehavior and hematoma volume after the intervention of breaking blood expelling stasis granules. The expressions of peroxidase proliferator-activated-receptor γ(PPARγ), CD11b and CD36 in the surrounding tissues of hematoma were detected by Western blot on the third day after the intervention. Results:After two weeks of treatment, the National Institutes of Health Stroke Scale (NIHSS) score and hematoma volume of the two groups decreased (all P<0.05), and the NIHSS score and hematoma volume of the observation group were significantly lower than those of the control group (all P<0.05). In addition, the changes of NIHSS score and hematoma volume in the observation group before and after treatment were significantly greater than those in the control group ( P<0.01). In animal experiments, the hematoma volume in the breaking blood expelling stasis group on the 14th day after ICH was significantly smaller than that in the ICH group [(9.8±4.9)mm 3 vs (17.6±6.4)mm 3,P<0.05], and the reduction of hematoma in the breaking blood expelling stasis group on the 7th and 14th day was significantly larger than that in the ICH group [(4.6±2.9)mm 3 vs (-2.1±1.6)mm 3, (14.3±3.8)mm 3 vs (4.2±2.8)mm 3, all P<0.01]. The percentage of right turn on 3rd, 7th and 14th day and the modified Neurological Severity Score (mNSS) on 7th and 14th day in the breaking blood expelling stasis group were lower than those in the ICH group (all P<0.05). Western blot analysis showed that the expressions of CD11b, CD36 and PPARγ in the breaking blood expelling stasis group on the third day after ICH were significantly higher than those in the ICH group (CD11b: 0.78±0.12 vs 0.49±0.11, P<0.05; CD36: 1.16±0.16 vs 0.80±0.11, P<0.05; PPARγ: 0.78±0.11 vs 0.37±0.10, P<0.01). Conclusions:Breaking blood expelling stasis can effectively accelerate intracerebral hematoma clearance and improve neurological outcome after ICH, and the mechanism maybe probably mediated by activating PPARγ and enhanced CD36, CD11b upregulation on microglia/macrophages, which resulting in facilitating erythrocyte endogenous phagocytosis.

Influencing factors of Toric intraocular lens postoperative rotation and its principles of treatment / 国际眼科杂志(Guoji Yanke Zazhi)

Correcting astigmatism safely and effectively has become a crucial part of modern cataract surgery due to the transformation of the surgery into a refractive procedure. The increased predictability and enhanced safety of Toric intraocular lens(IOL)implantation has made it the preferred method of correcting corneal regular astigmatism above 0.75D in cataract surgery. Toric IOL needs to be implanted in a precise axis position to achieve good astigmatism correction. A major cause of toric misalignment is postoperative rotation, which typically occurs soon after surgery. However, large axis misalignment will eliminate the astigmatism corrective effect of Toric IOL, even cause astigmatism in a new axis position. The factors responsible for IOL postoperative rotation are diverse. As a result, profound understanding of the factors is crucial for clinicians to minimize the rotation. Repositioning procedure is generally selected in case of significant rotation and the timing of the procedure is vital. This paper reviewed the influencing factors of IOL rotation postoperatively and its principle of treatment.

A study on the rapidly non-destructive detection method of Huoxiang Zhengqi oral liquid using near infrared spectroscopy / 药学学报

Based on near infrared spectroscopy and high performance liquid chromatography, this paper established the regression relationship between near infrared spectroscopy and index component content of Huoxiang Zhengqi oral liquid, so as to realize the rapid detection of index component content based on near infrared spectroscopy. Magnolol, honokiol and hesperidin were used as the quality indexes of Huoxiang Zhengqi oral liquid. After using the first derivative and normalization pretreatment method, characteristic variables were screened by CARS, and the correction model was finally established by partial least-squares regression (PLSR) method. The method accuracy was evaluated with the external validation, and the prediction results were tested for significance. The results indicated that when the near infrared spectrum was scanned through the bottle, the model's correlation coefficients of prediction (Rp) were higher than 0.99, the root mean square errors of the prediction models (RMSEP) were all less than 0.008 4, and the relative standard errors of prediction set (RSEP) were all less than 2.83%. There was no significant difference in the predicted results between these two kinds of model. The models established in the non-destructive way have good performance and high prediction accuracy. The rapid and nondestructive way has application value in the quality control of Huoxiang Zhengqi oral liquid.

Treatment of comminuted fracture of inferior pole of patella with locking suspension and vertical fixation with three steel wires / 中国骨伤

OBJECTIVE@#To explore the clinical application of lockedge suspension combined with three steel wires vertical fixation in comminuted fracture of inferior pole of patella.@*METHODS@#From August 2016 to May 2019, 23 patients with comminuted fracture of the lower pole of the patella, including 14 males and 9 females, were treated with lockedge suspension combined with three steel wires vertical fixation. The age ranged from 34 to 68 (55.0±1.2) years. One year after operation, the pain and function were evaluated by pain visual analogue scale(VAS) and knee flexion and extension range of motion, and the clinical efficacy was evaluated by Lysholm knee score standard.@*RESULTS@#All 23 patients were followed up for 12 to 14, with a mean of(13.0±0.5) months. One patient had skin irritation by the tail of the steel wire, and the rest had no postoperative complications such as incision infection, internal fixation loosening and fracture displacement. The fractures of 23 patients were healed, and the healing time was 10 to 14 weeks with a mean of(12.0±1.1) weeks. The VAS score decreased from 7.96±0.93 before operation to 0.83±0.65 one year after operation. The range of knee flexion and extension activities increased from(20.30±8.69) ° before operation to 1 year after operation(127.39±6.55) °. Lysholm knee score increased from 18.48±4.00 before operation to 96.09±4.91 one year after operation(P<0.05).@*CONCLUSION@#The treatment of comminuted fracture of the lower pole of patella by lockedge suspension combined with three steel wires vertical fixation has reliable fixation and high fracture healing rate. It can meet the requirements of rapid rehabilitation and functional exercise, and the early clinical effect is satisfactory.

Effect and Safety of Modified Buzhong Yiqitang on VEGF， IGF-1， TGF-β1， and Immune Function in Postoperative Patients with Non-small Cell Lung Cancer After Chemotherapy / 中国实验方剂学杂志

Objective:To explore the application value of modified Buzhong Yiqitang （BZYQT） in the treatment of postoperative patients with non-small cell lung cancer （Qi deficiency in lung and spleen） after chemotherapy， and to observe its effect on tumor angiogenesis， immune function， tumor indicators， and lung function indicators. Method:Ninety-six patients who were treated in the Kunming municipal hospital of traditional Chinese medicine from March 2018 to February 2020 due to postoperative chemotherapy for non-small cell lung cancer were selected and assigned into a control group （<italic>n</italic>=48， western medicine） and an observation group （<italic>n</italic>=48， western medicine+modified BZYQT） by the random number table. The curative efficacies were compared after the treatment. Result:After treatment， the serum levels of carcinoembryonic antigen （CEA）， cytokeratin 19 fragment 21-1 （CYFRA21-1）， serum insulin-like growth factor-1 （IGF-1）， vascular endothelial growth factor （VEGF）， and transforming growth factor（TGF）-<italic>β</italic>₁ in the observation group were lower than those in the control group （<italic>P</italic><0.05）， while the serum CD4⁺/CD8⁺，CD4⁺ cells， immunoglobulin G （IgG） levels， forced expiratory volume in one second （FEV₁），and FEV₁/forced vital capacity （FVC） in the observation group were higher than those in the control group （<italic>P</italic><0.05）. A significant difference was observed in the total response rate between the observation group ［56.25% （27/48）］ and the control group ［35.42% （17/48）］ （<italic>χ</italic>²＝4.191，<italic>P</italic><0.05）. For adverse reactions，the incidence of bone marrow suppression（<italic>χ</italic>²＝4.002）， gastrointestinal reaction （<italic>χ</italic>²＝7.069），and hepatic and renal injury （<italic>χ</italic>²＝5.151） was lower in the observation group than in the control group （<italic>P</italic><0.05）. Conclusion:For postoperative patients with non-small cell lung cancer （Qi deficiency in lung and spleen） after chemotherapy， western medicine combined with modified BZYQT could ameliorate immune function， promote pulmonary function recovery， improve clinical efficacy， and reduce the incidence of adverse reactions.

Clinical oberservation on rotator cuff suture threader and strapping suture in meniscus tear under arthroscopy / 中国骨伤

OBJECTIVE@#To explore clinical effect of arthroscopic meniscus tear strapping suture by rotator cuff suture threader.@*METHODS@#Forty patients with meniscus tear injury admitted from July 2015 to May 2019, including 27 males and 13 females, aged from 20 to 55 years old with an average of (36.0±1.4) years old. Menisci laceration was sutured with rotator cuff suture thread under arthroscopy. Postoperative complication was observed, Lysholm knee joint score before and after operation at 12 months were used to evaluate clinical effects, visual analogue scale (VAS) and range of knee flexion and extension were applied to evaluate recovery of pain and function.@*RESULTS@#All patients were followed up from 12 to 15 months with an average of (12.6±0.7) months.No complication such as joint effusion, suture failure occurred. Two patients occurred mild pain after activity without clinical physical abnormality, and 1 patient manifested moderate pain with joint space tenderness, the other rest without abnormal. Lysholm knee joint score was increased from (49.55±1.21) preoperatively to (98.95±0.42) at 12 months after operation, VAS score decreased from (5.18±0.78)preoperatively to (1.03±0.77) at 12 months after operation, and range of knee joint flexion and extension activity increased from (50.63±9.20)°preoperatively to (130.38±4.99)°after operation, and there were statistical differences in Lysholm knee joint score, VAS and range of knee joint flexion and extension activity (@*CONCLUSION@#Arthroscopic strapping suture by rotator cuff suture threading device applies to most meniscus injuries, including medial meniscus posterior horn tears, lateral meniscus body tears and lateral meniscus posterior horn tears. This technique meets the need of full-internal meniscus suture without specialmeniscus suture, and has advantages of convenient operation, less complications and good postoperative function.

Analysis of classical prescription Jinshui Liujun Jian based on ancient literature / 中国中药杂志

To provide the ancient literary evidence support for the clinical application and development of classical prescription based on systematical collection and analysis of the ancient Chinese medical literature containing Jinshui Liujun Jian, including its origin and development. Bibliometric analysis was used and information of Jinshui Liujun Jian in ancient Chinese medical literature was then collected for statistical analysis of formula compositions, main indications, dosage, preparation methods, etc. A total of 151 valid items of data were obtained from 48 ancient Chinese medicine books. Jinshui Liujun Jian was first recorded in Jingyue Quanshu written by ZHANG Jiebin. This prescription consisted of Rehmanniae Radix Praeparata, Angelicae Sinensis Radix, Pinelliae Rhizome, Citri Reticulatae Pericarpium, Poria and Glycyrrhizae Radix et Rhizome Praeparata cum Melle, and it was mainly used to treat the deficiency of lung and kidney, edema and excess production of phlegm, or Yin deficiency in the old, insufficient blood-qi, wind-cold evil, cough and disgusting, asthma and excessive phlegm. Doctors in later dynasties mostly followed the prescription compositions, dosages and indications in Jingyue Quanshu, and extended the clinical application of this prescription.

Analysis of differences in clinical characteristics between multifocal and multicentric breast cancer and unifocal breast cancer / 中山大学学报(医学科学版)

@#【Objective】 To explore the differences of clinical medicine ，magnetic resonance imaging（MRI）and pathology in multifocal and multicentric breast cancer（MMBC）and unifocal breast cancer（UBC）. 【Methods】 In this retrospective analysis，55 MMBC and 68 UBC patients with pathology confirmed from April 2016 to February 2018 were enrolled，and the characteristics and difference of routine pathological types，molecular subtypes and MR enhancement types were compared. The relationships between MMBC ，UBC and the methods of clinical treatment were studied by correspondence analysis（CA）.【Results】Significant difference was observed between routine pathological types of MMBC and UBC（P < 0.001）. The high grade invasive ductal carcinoma was more frequent in maximal lesions of MMBC than in UBC lesions，whereas there was no statistical correlation between molecular subtypes，molecular subtypes and MR enhancement types（P = 0.265，P = 0.152）. However，there was statistical difference in masses enhancement（P = 0.013）. CA showed that the molecular subtypes of MMBC and UBC were the key factors for clinical treatment. In addition ，HER- 2（+）and Luminal B type breast cancer showed high correlation with treatment method，while triple-negative showed low correlation with treatment method.【Conclusions】The pathology types of the maximal lesions of MMBC were less aggressive than UBC lesions. There was significant correlation between clinical treatment and molecular subtypes of MMBC and UBC. Therefore，individualized treatments are recommended on the basis of biological characteristics in both MMBC and UBC.

Correlation between CD8 + T cells' balance and prognosis for patients with diffuse axonal injury / 中国医师杂志

Objective To study the correlation between CD8 + T cells' balance and prognosis for patients with diffuse axonal injury(DAI).Methods To collect the 41 patients with DAI as observation group,with 33 cases light craniocerebral injury (LCI) patients and 35 healthy volunteers as control group.We detect the peripheral blood CD8 + CD28 + T cells and CD8 + CD28-T cell percentage,and calculate the ratio of both.The subjects working curve method (ROC) of.the above CD8 + T cells and its ratio were used to predict the prognosis of patients with DAI efficiency evaluation.Results (1) Compared with control group,CD8 + CD28 + T cells in patients with DAI and LCI increased significantly,and DAI was significantly higher than that of LCI group (P ＜ 0.05).Compared with control group,CD8 + CD28-T cells in patients with DAI were decreased,but no statistical difference was found between LCI and the control group,but DAI group was significantly lower than the LCI group (P ＜ 0.05).(2) CD8 + CD28-T cells and CD8 + CD28 +/CD8 + CD28-ratio were correlated with DAI diagnosis (P =0.003,0.000).When the percentage of CD8 + CD28-T cells =2.95％,the sensitivity of the diagnosis of DAI was 87.49％,86.21％;When the ratio of CD8 + CD28 +/CD8 + CD28-=7.39,the sensitivity was 92.63％,90.71％.(3) a total of 7 cases of patients died within 1 week after injury,CD8 + CD28 + T cells and CD8 + CD28 +/CD8 + CD28-ratio were significantly negative correlated with survival time (r =-0.739,-0.834,P =0.021,0.002),and CD8 + CD28-T cells were significantly positive correlated with survival time (r =0.782,P =0.006).Conclu sions Decreasing CD8 + CD28-T cells and higher CD8 + CD28 +/CD8 + CD28-ratio in patients with DAI immunological characteristics,especially the ratio (balance) is closely corrected with the diagnosis and prognosis of DAI.

Study on fingerprints of Huagai San-medicinal materials based on peak pattern matching / 中草药

Objective: To establish HPLC fingerprints of Huagai San and identificate the characteristic peaks ion and preparations- medicinal materials peak pattern matching. Methods: Similarity on 12 batches of Huagai San was estimated, and peak pattern matching of the original medicine was conducted. Sample extraction with methanol in ultrasonic condition was carried out. HPLC- DAD methods were developed, detection wave length was set at 280 nm, HPLC mobile phase consisted of acetonitrile-0.2% phosphate acid aqueous solution, volume flow-rate was delivered at 0.6 mL/min, and column temperature was kept at 30℃. Cluster analysis of 12 batches of Huagai San was analyzed by SPSS 19.0. Results: The results showed that 26 common peaks were defined, the peaks No. 4, 5, 10, 17, 18, 20, and 21 were ephedrine hydrochloride, pseudoephedrine hydrochloride, chlorogenic acid, liquiritin, glycyrrhizic acid, narirutin, and hesperidin, respectively, and peak pattern matching showed that Huagai San had 6, 3, 6, 1, 4, and 1 matching peaks with Ephedrae Herba, Mori Cortex, Citri Reticulatae Pericarpium, Armeniacae Semen Amarum, Glycyrrhizae Radix et Rhizoma, and Perillae Fructus, respectively. The fingerprint similarity among the samples was all over 0.960. All the samples were classified into three categories. Conclusion: Similarity evaluation combined with the peak matching of fingerprint for Huagai San could provide the scientific methods for its identification and quality control.

Antitumor Effect of Ganoderma lipsiense Extract on Triple-negative Breast Cancer Model Mice and Mechanism Study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the inhibitory effect and mechanism of Ganoderma lipsiense extract (GLE) on the growth of triple-negative breast cancer (TNBC) cell line MDA-MB-231-HM in a mouse model.</p><p><b>METHODS</b>The mouse model of TNBC was established by subcutaneous injection of 1.5 x 10(6) of MDA-MB-231-HM cells into BALB/c-nu mouse. Twenty successfully modeled mice were divided into the GLE group and the negative control group according to random digit table, 10 in each group. GLE (0.2 mL 100 mg/mL) was peritoneally injected to mice in the GLE group, while equal dose of normal saline was peritoneally injected to mice in the negative control group. The medication was administered once per 3 days and discontinued after 45 days. The CD34 expression was detected using immunohistochemical assay for counting microvessels. Meanwhile, expressions of thrombospondin 1 (TSP-1) and cyclin D1 were detected using immunohistochemical assay.</p><p><b>RESULTS</b>The average weight was obviously lower in the GLE group than in the negative control group [(0.33 ± 0.16) g vs (0.68 ± 0.37)g, P < 0.05]. The tumor inhibition rate was 51.4% in the GLE group. The volume of transplanted tumor was obviously lesser in the GLE group than in the negative control group (P < 0.05). Results of immunohistochemical staining showed, the microvessel density (MVD) under every field was (20.7 ± 2.1), TSP-1 positive cell count was (66.2 ± 9.2), cyclin D1 positive cell count was (33.8 ± 16.4) in the GLE group, and they were 34.0 ± 2.0, 24.0 ± 6.6, and 168.2 ± 32.6, respectively in the negative control group. There was statistical difference in all indices between the two groups (P < 0.05).</p><p><b>CONCLUSION</b>GLE could inhibit malignant proliferation of tumor cells by suppressing angiogenesis of blood vessels in tumor tissues and regulating cell cycles, thereby inhibiting TNBC.</p>

Bridging fixation with locking plate for the treatment of subtrochanteric femoral fractures of Seinsheimer type V / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the clinical effect of bridging fixation with locking plate for the Seinsheimer type V subtrochanteric femoral fracture.</p><p><b>METHODS</b>From March 2009 to September 2014,18 cases of Seinsheimer type V subtrochanteric femoral fracture were treated by open reduction and bridging fixation with locking plate through proximal and distal approach including 16 males and 2 females with an average age of 41 years old ranging from 22 to 67 years old. Among them, 12 cases caused by traffic accident, 5 cases by falling, 1 case by heavy aboving. All cases were fresh and closed fractures. Time between injury and operation was from 4 to 9 days with an average of 6.2 days. Of them, 11 cases were fixed with reverse LISS and the other 7 cases were fixed with anatomical locking plates of proximal femur.</p><p><b>RESULTS</b>The mean time of operation was 110 min (ranged from 90 to 155 min). The mean blood loss during operation was 425 ml (ranged from 350 to 650 ml) and 16 cases got blood transfusion which was meanly 300 ml. The mean hospital time was 14 days (ranged from 12 to 18 days). The mean duration of followed up was 11.8 months (ranged from 8 to 22 months). The mean time of bone union was 6.6 months (ranged from 5 to 8 months). There was not any complication such as infection, implant failure, hip varus, external rotation deformity of low limb or fat embolism. The Sanders hip scores were 53.22 ± 6.48, the result was excellent in 12 cases and good in 6 cases at the last follow-up.</p><p><b>CONCLUSION</b>Under the principle of biological osteosynthesis, treatment of Seinsheimer type V subtrochanteric femoral fracture with bridging locking plate fixation has such advantages as high mechanism, less interference of blood supply, stable fixation and little complication. It is a safe and idea way for the treatment of the Seinsheimer type V subtrochanteric femoral fracture.</p>

Case-control study on close reduction combined with minimally invasive percutaneous plate osteosynthesis for the treatment of distal fracture of tibial shaft / 中国骨伤

<p><b>OBJECTIVE</b>To compare the effects of close reduction combined with minimally invasive percutaneous plate osteosynthesis for distal fracture of tibial shaft.</p><p><b>METHODS</b>From March 2009 to May 2013, there were 124 patients (89 males and 35 females, 30 of them were injured in a traffic accident and 94 were falling down in daily life, the ages ranging from 21 to 81 years old) who suffered from distal fracture of tibial shaft. Sixty-six patients (48 males and 18 females, 45 cases of type A, 12 cases of type B and 9 cases of type C) were treated with close manipulative reduction combined with minimally invasive percutaneous plate fixation. After close reduction, a minimal incision was made and a anatomic plate was inserted just along the medial tibia periostea, and then the fracture was fixed without fracture exposure. The other 58 patients (41 males and 17 females, 41 cases of type A, 10 cases of type B and 7 cases of type C) were treated with conventional open reduction and internal fixation. Length of the incision, operating time, early postoperative pain (recorded using the Visual Analog Scale score) and the outcome results (recorded using the evaluation standard of Johner-Wruhs) were compared.</p><p><b>RESULTS</b>The length of incision was meanly (7.34 ± 1.42) cm in MIPPO group and (21.82 ± 2.35) cm in ORIF group; operation time was (44.48 ± 10.00) min in MIPPO group and (59.42 ± 11.84) min in ORIF group. The postoperative radiographs verified good position of all screws and satisfactory bone fracture reduction in both groups. All the patients were followed up, and the duration ranged from 10 to 24 months (mean 15.2 months) in both groups. In MIPPO group,only one patient had delayed union and got union after Chinese herb therapy. The other 65 patients got bony union during 15 to 20 weeks. While in ORIF group, 3 patients suffered from nonunion and received reoperation with bone grafting, and 4 patients got bone infection.</p><p><b>CONCLUSION</b>Treatment of distal fracture of tibial shaft, combined with close reduction and MIPPO technique, has the advantages such as less invasion, less damage of blood supply, simplified procedure of operation and higher union rate, which is an ideal methods and is accordant to the biological demand.</p>

Protective effects of WR2721 on early bone marrow hematopoietic function in mice exposed to 6.5 Gy of (60)Co γ-rays / 中国实验血液学杂志

The aim of this study was to investigate the effect of WR2721(amifostine) against bone marrow hematopoietic damage of mice exposed to 6.5 Gy of (60)Co-γ ray. A total of 60 C57/BL6J mice was divided into 3 groups:normal group (mice were injected with physiological salt solution), irradiation group (mice were injected with physiologic salt solution before irradiation) and WR2721 group (mice were injected with WR2721 before irradiation). The WBC, neutrophil (Neut), Plt and RBC levels in peripheral blood of 3 group mice were counted within 60 days after irradiation; the bone marrow nuclear cells (BMNC) were counted at 2 and 24 hours after irradiation; the hematopoietic stem/progenitor cell (LK/LSK) level and colony formation capability were detected by flow cytometry at 2 and 24 hours after irradiation. The results indicated that the counts of WBC and neut at 4 and 18 days, Plt at 7-18 days and RBC at 10-30 day after irradiation in WR2721 group were higher than those in irradiation group (P < 0.05); the BMNC, LSK and LK levels obviously increased at 24 hours after irradiation (P < 0.05), the CFU-GEMM, CFU-GM, CFU-MK BFU-E and CFU-E all significantly increased at 2 and 24 hours after irradiation (P < 0.01), as compared with irradiation group. It is concluded that WR2721 can effectively alleviate early hematopoietic damage and promote the fast recovery of peripheral blood cells in mice exposed to γ-ray, suggesting that the WR2721 has significant radioprotective effect on hematopoietic system.

Identifying interacting proteins of a Caenorhabditis elegans voltage-gated chloride channel CLH-1 using GFP-Trap and mass spectrometry / 生理学报

Chloride channels belong to a superfamily of ion channels that permit passive passage of anions, mainly chloride, across cell membrane. They play a variety of important physiological roles in regulation of cytosolic pH, cell volume homeostasis, organic solute transport, cell migration, cell proliferation, and differentiation. However, little is known about the functional regulation of these channels. In this study, we generated an integrated transgenic worm strain expressing green fluorescence protein (GFP) fused CLC-type chloride channel 1 (CLH-1::GFP), a voltage-gated chloride channel in Caenorhabditis elegans (C. elegans). CLH-1::GFP was expressed in some unidentified head neurons and posterior intestinal cells of C. elegans. Interacting proteins of CLH-1::GFP were purified by GFP-Trap, a novel system for efficient isolation of GFP fusion proteins and their interacting factors. Mass spectrometry (MS) analysis revealed that a total of 27 high probability interacting proteins were co-trapped with CLHp-1::GFP. Biochemical evidence showed that eukaryotic translation elongation factor 1 (EEF-1), one of these co-trapped proteins identified by MS, physically interacted with CLH-1, in consistent with GFP-Trap experiments. Further immunostaining data revealed that the protein level of CLH-1 was significantly increased upon co-expression with EEF-1. These results suggest that the combination of GFP-Trap purification with MS is an excellent tool to identify novel interacting proteins of voltage-gated chloride channels in C. elegans. Our data also show that EEF-1 is a regulator of voltage-gated chloride channel CLH-1.

Genetic evolution analysis of matrix protein 2 gene of avian influenza H5N1 viruses from boundary of Yunnan province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein(M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012.</p><p><b>METHODS</b>A total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos,myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT-PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains.</p><p><b>RESULTS</b>A total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained from 30 positive samples and were divided into 3 distinct clades or sub-clades(1.2.1, 1.2.2 and 2) by phylogenetic analysis, 5, 7 and 2, respectively. The M2 genes and Hemagglutinin(HA) genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains.</p><p><b>CONCLUSION</b>The M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2.2 had become dominant epidemic strain in this region.</p>

Preparation of paeoniflorin microcapsule and its in vitro release behavior / 中草药

Objective: To optimize the preparation technology of paeoniflorin microcapsule (PM), and to study its behavior of in vitro release. Methods: Using encapsulation efficiency and drug loading as indicators, the PM were prepared by complex agglutination method, the preparation technology of PM was optimized by Doehlert design. The dissolution volume of PM within 20 h and the release curve were measured by HPLC, afterwards its morphology and particle size were studied by electron microscopy. Results: The encapsulation efficiency and drug loading were significantly related to the ratio of coating material, stirring speed, and pH value. The optimal conditions were as follows: the ratio of coating material to paeoniflorin was 4.3:1, the stirring speed was 305 r/min, and the pH value was 4.0. The obtained microcapsules were smooth round capsule-shaped, with non-adhesions and uniform particle size, the encapsulation efficiency was up to 83.81%, the drug loading was 24.24%, and the microcapsule diameter was below 200 μm, with sustained-release effect. Conclusion: The complex agglutination method is simple and reliable to prepare PM, and the product is stable. As a new formulation, microcapsule has a broad prospect for development.

Genetic evolution of non-structural gene among avian influenza H5N1 viruses isolated from the boundary of Yunnan province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To elucidate the characteristics of variation and the genetic evolution of non-structural protein (NS1, NS2) genes related to avian influenza subtype H5N1 viruses isolated from the boundary region of Yunnan province.</p><p><b>METHODS</b>Swab samples were collected from foreign poultry and wild birds in the boundary regions of Yunnan province and screened by H5/N1 subtype-specific multiplex RT-PCR. The NS segment of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis on those available NS1, NS2 genes were performed with sequences of the known reference strains.</p><p><b>RESULTS</b>71 positive samples were identified from 1240 samples, with the positive rate as 5.72%. Fourteen different NS segment sequences were obtained from 30 representative positive samples and could be divided into 3 distinct clades or sub-clades (I-1, I-2 and II), by phylogenetic analysis. The NS1/NS2 genes and Hemagglutinin (HA) genes of H5N1 viruses from the boundary regions of Yunnan province showed different relationships regarding the characteristics on genetic evolution. The substitution or mutation of key amino acids sites had been noticed in the nuclear location signal domains, effect domain, and other pathogenicity markers.</p><p><b>CONCLUSION</b>NS genes of H5N1 subtype viruses in boundary region of Yunnan province showed genetic divergence and the virus of clade I-2 and II had become dominant epidemic strains in this region since 2010.</p>

Genetic diversification of avian influenza H5N1 virus in boundary areas of Yunnan province / 中华流行病学杂志

Objective To elucidate the genetic diversifications of avian influenza subtype H5N1 viruses in the boundary regions of Yunnan province during 2009 to July,2011.Methods Swab samples were collected from foreign poultry and wild birds in boundary regions of Yunnan province during 2009 to July,2011 and tested by H5/N1 subtype-specific multiplex RT-PCR.The HA genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD 18-T vectors for sequencing.Both alignment and phylogenetic analysis were performed with sequences of the known reference strains.Results Fifteen different HA sequences were obtained from 36 representative positive samples and could be divided into 2 distinct Clades (2.3.2 and 2.3.4).Through phylogenetic analysis,Clade 2.3.2 and 2.3.4 could then be further divided into 3 ( Ⅱ -1 to Ⅱ -3 ) and 2 smaller clades ( Ⅰ -1 and Ⅰ -2),respectively.The viruses of Clade 2.3.2 Ⅱ -1 and Ⅱ-2 were new variant strains of H5N 1 virus.The cleavage sites of HA from positive samples all possessed molecular characterization of highly pathogenic avian influenza virus.Mutation of key amino acids had been found among receptor binding sites,potential glycosylation sites,neutralizing epitopes and others.Conclusion It seemed evident that the H5N1 subtype viruses showed genetic diversifications and had undergone the evolution progress of multi-clade (2.3.2,2.3.4) to single caide (2.3.2) in the boundary regions of Yunnan province,during 2009 to July,2011.

Reduced brain-derived neurotrophic factor expression in cortex and hippocampus involved in the learning and memory deficit in molarless SAMP8 mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The molarless condition has been reported to compromise learning and memory functions. However, it remains unclear how the molarless condition directly affects the central nervous system, and the functional consequences on the brain cortex and hippocampus have not been described in detail. The aim of this study was to find the molecular mechanism related with learning and memory deficit after a bilateral molarless condition having been surgically induced in senescence-accelerated mice/prone 8 (SAMP8) mice, which may ultimately provide an experimental basis for clinical prevention of senile dementia.</p><p><b>METHODS</b>Mice were either sham-operated or subjected to complete molar removal. The animals' body weights were monitored every day. Learning ability and memory were measured in a water maze test at the end of the 1st, 2nd, and 3rd months after surgery. As soon as significantly prolonged escape latency in the molarless group was detected, the locomotor activity was examined in an open field test. Subsequently, the animals were decapitated and the cortex and hippocampus were dissected for Western blotting to measure the expression levels of brain-derived neurotrophic factor (BDNF) and the tropomyosin related kinase B (TrkB), the high affinity receptor of BDNF.</p><p><b>RESULTS</b>Slightly lower weights were consistently observed in the molarless group, but there was no significant difference in weights between the two groups (P > 0.05). Compared with the sham group, the molarless group exhibited lengthened escape latency in the water maze test three months after surgery, whereas no difference in locomotor activity was observed. Meanwhile, in the cortex and hippocampus, BDNF levels were significantly decreased in the molarless group (P < 0.05); but the expression of its receptor, TrkB, was not significantly affected.</p><p><b>CONCLUSION</b>These results suggested that the molarless condition impaired learning and memory abilities in SAMP8 mice three months after teeth extraction, and this effect was accompanied by significantly reduced BDNF expression in the cortex and hippocampus.</p>