Visual Detection of Plant Viral RNA Based on Gold Nanoparticles and Hybrid Chain Reaction / 中国生物化学与分子生物学报

The harm of plant virus disease is serious, which significantly restricts the sustainable development of agriculture and can cause huge economic losses. Monitoring plant health and early detection of viral pathogens are essential to reduce the spread of disease. Therefore, in order to realize the early detection of plant viral diseases in the field, a sensitive, specific and efficient colorimetric visual technique for plant viral RNA was designed by combining the colorimetric method based on gold nanoparticles (AuNPs) and hybrid chain reaction (HCR). In this study, tobacco mosaic virus (TMV) was used as a model to design two hairpin structures H1/ H2 with single-stranded tails based on TMV-specific conserved fragments. TMV could open the hairpin structure to alternately form long double-straight DNA. The binding difference between AuNPs and two nucleic acid states before and after HCR reaction resulted in colorimetric signal generation, thus realizing visual detection of TMV. After optimizing the concentration of Tris-HAc, the concentration of hairpin structure and the reaction time of HCR, the best detection conditions were obtained. The sensitivity and specificity of the technique were analyzed and real samples were tested under optimal conditions. The results showed that the absorbance ratio of AuNPs (A

Identification and Analysis of Complete Genomic Sequences of Soybean Mosaic Virus Isolates from Atractylodes macrocephala Koidz / 中国生物化学与分子生物学报

Soybean mosaic virus (SMV) is a critical pathogen that reduces the soybean yield and seed quality worldwide, and SMV has a restricted natural host range. In this study, we used sequence-independent amplification (SIA) methods to identify the viruses that may cause the Atractylodes macrocephala Koidz disease. Results revealed that there is SMV in diseased Atractylodes macrocephala leaves, and we named this isolate as SMV-Am. To further characterize the genomic structural and phylogenetic relationships of SMV-Am, genomic dsRNA was extracted, and the genomic sequence was amplified by RT-PCR and RACE. The results of bioinformatics analysis showed that the genomic RNA of SMV-Am is 9587 nucleotides in length, which conforms to the typical characteristics of Potyvirus. According to the sequence alignment of complete nucleotide sequences, SMV-Am showed the highest level of nucleotide homology and amino acid sequences to SMV-Liaoning, 96. 57% and 98. 86%, respectively. In addition, phylogenetic analysis based on the nucleotide sequences of other SMV isolates of SMV-Am revealed that SMV-Am was most closely related to SMV-Liaoning. Then, the SMV-Am protein was further analyzed by I-TASSER and PyMOL software, revealing that the key amino acid mutations led to the structural changes of P1, HC-Pro, P3, 6K2, NIa-pro and NIb proteins, with P1 the most obvious. Finally, recombination has been detected at the position of 6560-8950 nucleotides, the main parent is the isolate SMV-XFQ012 (accession number KP710875. 1), and the secondary parent is isolate SMV-pCB301-SC15 (accession number MH919386. 1). This study is the first report of SMV in Atractylodes macrocephala Koidz, and these results are expected to provide a scientific basis for the prevention and control of SMV infection on Atractylodes macrocephala Koidz.

Clinical study on new risk assessment and prediction system for early osteonecrosis of the femoral head / 中国骨伤

OBJECTIVE@#To establish a risk assessment and prediction system for early osteonecrosis of the femoral head (ONFH) in order to predict the collapse risk.@*METHODS@#The risk assessment system for early necrosis and collapse of femoral head was established based on the combination of Steinberg stage, ABC typing and the proportion of the proximal sclerotic rim. Firstly, Steinberg stage system was applied. ABC typing was applied to predict risk in stage I, type C was risk free, type B was low risk, type A and type BC were medium risk, type A-C and type AB were high risk. The classification of proximal sclerotic rim was first applied when the Steinberg stage was Ⅱ-Ⅲ, and type 2 was expected to be low risk. If the classification of proximal sclerotic rimwas type 1, then the ABC typing was applied, type C was risk-free, type B was low risk, type A and type BC were medium risk, type A-C and type AB were high risk. According to this prediction system, the collapse risk of femoral head in 188 cases(301 hips) were predicted by retrospective analysis. All the hips were enrolled at the out-patient department of orthopedic in Guang'anmen Hospital attached to China Academy of Chinese Medical Science. The consistency of the prediction results of three doctors and one doctor at different times were evaluated.@*RESULTS@#Among them, 136 cases were male, 52 were female. 75 cases were single hip, 113 were double hip. The age of the patients wa 19 to 64(42.61±12.07) years. The natural course of disease was 0.33 to 5.00(3.62±1.93) years. 206 hips in 301 hips had collapsed, with a collapse rate of 68.44%. In the risk-free group, none hip had collapsed, with a collapse rate of 0%. In the low-risk group, 9 hip in 91 hips had collapsed, with a collapse rate of 9.89%. In the medium-risk group, 12 hip in 19 hips had collapsed, with a collapse rate of 63.16%. And in the high risk group, 185 hips in 190 hips had collapsed, with a collapse rate of 97.37%. They were significantly differences in their collapse rate (@*CONCLUSION@#The risk assessment and prediction system for early ONFH selects different methods to predict the risk of collapse according to the imaging characteristics of different stages, which is combines with the comprehensive assessment of multiple risk factors. The system is applicable to a wide range, simple operation and convenient for clinical application.

Establishment and Optimization of a Method for Rapid Extraction of Plant Virus RNA Based on Functional Magnetic Beads / 中国生物化学与分子生物学报

Plant virus diseases are one of the major diseases restricting erop production.Timely identification of their pathogen and development rules is the prerequisite for effective control of their large- scale spread.However, long cycle, tedious steps and strict detection environment were the disadvantages existing in the detection technology of plant virus disease.In this study, Tobacco Mosaic virus (TMV) was used as a model to he extract UNA based on CMBs-ACPtmv , which was design based on the principle of complementary base pairing.Meanwhile, the experimental conditions were optimized and analyzed, including the preparation conditions of functionalized magnetic beads, the reaction conditions during extraction, and the sensitivity, stability and other properties of the method.The results showed the ability to capture RNA of CMBs-ACPtmv were best when prepared with 4 fxmol capture probe (ACPTMV ) and 0.08 mg carboxyl magnetic beads (CMBs) ; After 3 min of extraction, CMBs-ACPtmv has the best RNA extraction effect, but when the extraction temperature of CMBs-ACPtmv was changed, its extraction capacity showed no significant change; In the comprehensive performance evaluation, the sensitivity of CMBs-ACPjjjv can reach 2.5 ng/fxL, and the detection stability is good.Compared with conventional RNA extraction technology, CMBs-ACPimv has outstanding advantages in detection time and sample consumption.The functional magnetic beads extraction method established in this study is fast, safe and simple.It can achieve rapid extraction of plant virus RNA with simple equipment, which has a broad application prospect.

Analysis of influencing factors the postoperative axial symptoms of cervical single open-door laminoplasty / 中国骨伤

OBJECTIVE@#To explore the influencing factors and possible mechanism of axial symptoms(AS) after C₃-C₇ single open-door laminoplasty in patients with chronic compression cervical myelopathy.@*METHODS@#The clinical data of 32 patients with multi-segment chronic compression cervical cord disease treated by C₃-C₇ single open-door laminectomy from May 2012 to July 2016 were retrospectively analyzed. Including cervical spondylotic myelopathy of 14 cases, developmental cervical stenosis complicated with cervical myelopathy of 8 cases, ossification of posterior longitudinal ligament(OPLL) of 10 cases. There were 17 males and 15 females, aged from 47 to 82 years old with an average of 57.46 year, the course of disease was 5 to 35 months with an average of 22.4 months. The opening angle(OA), cervical curvature angle(CA), preoperative spinal cord compression rate(PSCR) and postoperative spinal cord shift (PSCS) were recorded. After 2 weeks of surgery, determining whether occurred an AS condition according to the AS assessment criteria, the patients were divided into a axial symptom group and a non-axial symptom group, the general data and imaging parameters of the two groups were compared and the factors that may be postoperative AS were analyzed by binary Logistic regression analysis.@*RESULTS@#At 2 weeks after operation, 13 patients occurred AS. There was no significant difference in gender, age and course of disease between axial symptom group and a non-axial symptom group (>0.05). In axial symptom group, OA was(36.76±9.35)°, CA was(11.53±4.36)°, PSCR was(27.83±1.72)%, PSCS was (3.17±0.81) mm, while in non-axial symptom group, above items were (33.03±10.52)°, (7.71±4.73)°, (25.16±3.59)%, (2.43±0.95) mm, respectively, there was significant difference in CA, PSCR, PSCS between two groups(0.05). The results of the binary Logistic regression analysis of 3 parameters(OA, PSCR, PSCS) and AS showed OA and PSCR were eliminated in dependent variables, and the partial regression coefficient of PSCR was 0.311, and =0.031.@*CONCLUSIONS@#CA, PSCR, and PSCS are related influencing factors of AS, and PSCS is a high risk factor for AS. C₄,₅ nerve traction caused by posterior spinal movement, postoperative dural self-expansion causes greater traction of the spinal cord, excessive deformation of the cervical spinal cord causes autonomic nerve damage or necrosis that dominates blood vessels may be the pathogenesis of AS, but this is only a theoretical inference, and further improved experiment is necessary to verify it in the future.

Analysis of correlative factors of spinal cord posterior displacement after single door laminoplasty / 中国骨伤

OBJECTIVE@#To observe the open angle (OA), cervical curvature angle (CA), preoperative spinal cord compression rate(PSCR), postoperative spinal cord shift (PSCS) in patients with chronic compressive cervical myelopathy undergoing C3-7 single open laminoplasty, and to explore the possible mechanism and influencing factors of postoperative average spinal cord drift, so as to provide objective basis for predicting PSCS.@*METHODS@#From May 2012 to July 2016, 32 patients with multi-segmental chronic compressive cervical myelopathy who underwent single-door laminoplasty in our department were analyzed retrospectively, including 14 cases of cervical spondylotic myelopathy, 8 cases of developmental cervical spinal stenosis with cervical myelopathy, and 10 cases of ossification of posterior longitudinal ligament. The OA of cervical spine was measured on CT, the CA was measured on X-ray, the PSCR and PSCS were measured on MRI. The patients were divided into two groups according to PSCS(group A>=2.5 mm, group B0.1), and the partial regression coefficients of OA and PSCR were 0.113 and 0.059 respectively.@*CONCLUSIONS@#PSCS is the result of OA, CA and PSCR, among which PSCR has the most important influence, OA is the second, CA is the least. PSCS can be predicted by 0.059×OA+0.113×PSCR-2.266 equation, which provides a theoretical basis for preoperative evaluation of spinal cord decompression after surgery.

Claudin-3 expression in colorectal carcinoma and its significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of claudin-3 in colorectal carcinoma and its association with the occurrence, progression and prognosis of colorectal cancer.</p><p><b>METHODS</b>Forty surgical specimens of colorectal carcinoma and 22 adjacent normal tissues resected between October, 2010 and January, 2013 at Nanfang Hospital were examined for claudin-3 expression using immunohistochemistry, which was analyzed in association with the clinicopathological parameters and the survival of the patients.</p><p><b>RESULTS</b>Claudin-3 was expressed mainly on the cell membrane, and its positivity rate was significantly higher in cancer tissues than in normal tissues (92.50% vs 59.09%, P<0.05). In 13 cases claudin-3 expression was detected in both the cancer tissues and adjacent normal tissues with average expression scores of 4.538 and 3.269, respectively (P<0.05). In the cancer tissues, the strongly positive expression rate was significantly higher in poorly differentiated tissues (85.71%) than in well (21.43%) and moderately (36.48%) differentiated tissues (P<0.05), and was higher in cases with lymph node metastasis than in those without (61.11% vs 22.72%, P<0.05). The strongly positive expression rate of claudin-3 was not correlated with the patients'age, gender, tumor location or tumor size (P>0.05). Of the 33 cancer patients followed up, 14 had a postoperative survival time no longer than 3 years and 19 had longer survival time, and their average expression scores differed significantly (4.50 vs 3.526, P<0.05).</p><p><b>CONCLUSION</b>Claudin-3 is over-expressed in colorectal cancer tissues, and its high expression may promote the occurrence and progression of colorectal cancer. Claudin-3 may serve as a molecular biomarker for early diagnosis and prognostic evaluation.</p>

Pharmacokinetics of α-asarone after intranasal and intravenous administration with PLA-α-asarone nanoparticles / 中国中药杂志

PLA-α-asarone nanoparticles were prepared by using organic solvent evaporation method, and their in vivo distribution and brain targeting after intranasal administration were studied as compared with intravenous administration. The results showed that brain targeting coefficient of PLA-α-asarone nanoparticles after intranasal and intravenous administration was 1.65 and 1.16 respectively. The absolute bioavailability, brain-targeting efficiency and the percentage of nasal-brain delivery of PLA-α-asarone nanoparticles were 74.2%, 142.24 and 29.83%, respectively after intranasal administration. The results of fluorescence labeling showed that the fluorescent intensity of coumarin-6 in the brain tissue was the highest after intranasal administration of PLA-α-asarone fluorescent nanoparticles, achieving the purpose of brain-targeted drug delivery. The fluorescent intensity of coumarin-6 in liver tissue after intravenous administration of PLA-α-asarone nanoparticles was much higher than that after intranasal administration, indicating that intranasal administration of PLA-α-asarone nanoparticles could decrease drug-induced hepatotoxicity. In addition, the fluorescent intensity of coumarin-6 in lung tissue was weaker after intranasal administration, which solved the shortcomings of intranasal administration of α-asarone dry powder prepared by airflow pulverization method. In vivo studies indicated that PLA-α-asarone nanoparticles after intranasal administration had a stronger brain targeting as compared with intravenous administration.

Study on the Distribution of Constitutions of Chinese Medicine in Patients with Osteonecrosis of Femoral Head / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the distribution of constitution types of Chinese medicine (CM) in patients with osteonecrosis of femoral head (ONFH).</p><p><b>METHODS</b>Totally 130 ONFH patients were recruited. Constitution types of CM were identified in all patients. Distribution features of constitution types of CM in ONFH patients were observed. The differences of distribution in gender, age, single or bilateral hips, course of disease, staging, cause, and region were also analyzed.</p><p><b>RESULTS</b>Seventy patients were of complicated constitutions, while 60 patients were of single constitution. Among the 60 single constitution cases, yang-deficiency constitution [18 (13.9%)], damp-heat constitution [10 (7.7%)], blood-stasis constitution [7 (5.4%)], and qi-deficiency constitution [7 (5.4%)] were mainly distributed. Of the complicated constitutions, yang-deficiency dominated constitution occupied the top ratio [30 (23.1%)], followed by blood-stasis dominated constitution [15 (11.5%)], damp-heat dominated constitution [9 (6.9%)]. By putting them together, yang-deficiency constitution occupied the top constitution of CM [48 (36.9%)], followed by blood-stasis constitution [ 22 (16.9%)] and damp-heat constitution [19 (14.6%)]. The aforesaid three constitutions accounted for 68.5% of the total. There were no statistical distribution differences in gender, age, single or bilateral hips, course of disease, staging, or cause.</p><p><b>CONCLUSION</b>Yang-deficiency constitution, damp-heat constitution, and blood-stasis constitution were liable constitutions of CM in ONFH patients.</p>

Effects of bm47 deletion on viral replication and transcription of Bombyx mori nucleopolyhedrovirus / 病毒学报

Bombyx mori nucleopolyhedrovirus (BmNPV) bm47 gene is found in all sequenced lepidopteran nucleopolyhedroviruses (NPVs). It is one of the core genes of NPVs. However, the role of bm47 in the biological cycle of NPV remains unknown. In this study, the Red recombination system was used to knock out bm47 from BmNPV to construct bm47-ko-Bacmid in E. coli BW25113 system. Then bm47 gene was introduced back to the viral genome using the Bac-to-Bac system to create the repair virus bm47-re-Bacmid. TCID50 assay and real-time PCR (qPCR) were used to evaluate the effects of bm47 deletion on viral DNA replication, gene transcription, and protein expression. qPCR results showed that bm47 knock-out had no significant effect on viral DNA replication. However, the qPCR results showed that bm47-ko-Bacmid significantly decreased the transcription levels of early gene lef-3, late gene vp39, and very late gene p10 at 48 h and 72 h after viral transfection of BmN cells (P < 0.05). This work will provide a foundation for further studies on the biological function of BmNPV bm47 in viral replication and transcription.

Expression of microRNA-223 in lymphocytic leukemia cells and its action mechanism / 中国实验血液学杂志

This study was aimed to investigate the expression level and mechanism of microRNA-223 and LMO2 in acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL) cells and the mechanism. MicroRNA-223 mimics was transfected to increase the expression of MicroRNA-223 in the lymphocytes sorted by ficoll separation from the bone marrow mononuclear cells (BMMNC) of ALL and CLL patients. MicroRNA-223 inhibitor was transfected to decrease the expression of the MicroRNA-223 in the lymphocytes of normal controls. Then the expression of the MicroRNA-223 and LMO2 in transfected lymphocytes before and after cultivating for 72 hours were detected by RT-PCR, the apoptosis and cell cycle of these cells were measured by flow cytometery. The results indicated that before the transfection, the expression of MicroRNA-223 in ALL and CLL cells was (433.11 ± 144.88), which was significantly lower than that in the normal lymphocyte (949.59 ± 267.39); the expression of LMO2 was (807.10 ± 238.41), which was significantly higher than that in the normal lymphocytes (455.32 ± 176.83) (P < 0.05); after the transfection, the expression of MicroRNA-223 was (571.86 ± 142.00) in ALL and CLL cells, which was significantly higher than that before transfection (P < 0.05), but the expression of LMO2 was significantly lower than that before transfection (651.97 ± 230.12) (P < 0.05); in the normal control the expression of MicroRNA-223 obviously decreased (646.32 ± 172.93) (P < 0.05), the expression of LMO2 was significantly increased (541.27 ± 158.86.2) (P < 0.05). After transfection, the cell cycle G1/G2 phase and apoptosis changed in ALL and CLL cells. Before transfection the cell ratio in cell cycle G1/G2 phase was (94.75 ± 3.15)%, the cell ratio in S phase was (5.14 ± 3.12)%; after transfection the cell ratio in cell cycle G1/G2 phase was (97.03 ± 2.08)% and obviously increased (P < 0.05), the cell ratio in S phase was (2.97 ± 2.08)% and significantly decreased (P < 0.05). Before transfection the apoptosis rate was (54.47 ± 8.72)%, and obviously was higher than that after transfection (60.48 ± 8.81)%. And in the normal control, the cell ratio in G1/G2 phase was significantly higher than that after transfection [(96.73 ± 2.26)%, (94.55 ± 2.77)%, P < 0.05)], and the cell ratio in S phase was significantly increased [(3.25 ± 2.26)%, (5.45 ± 2.77)% (P < 0.05)]. The apoptotic rate in the ALL and CLL patients was significantly higher than that after the transfection [(54.47 ± 8.72)% vs (60.48 ± 8.81)%, respectively (P < 0.05)]. The apoptotic rate in the normal control was significantly lower than that after the transfection [(59.02 ± 10.20)%, (51.96 ± 10.20)%, respectively (P < 0.05)]. It is concluded that the expression of MicroRNA-223 decreases, and the expression of LMO2 increases in lymphocytic leukemia cells which leads to the lymphocytes over-proliferation and abnormal apoptosis, thus may be one of pathogenesis in lymphocytic leukemia.

Impact of chronic kidney disease on serum tumor markers concentrations / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Serum tumor markers have always been of clinical importance in the diagnosis, monitoring disease progression and therapy efficacy for patients with malignant diseases. However, elevated serum tumor markers are found in some benign conditions, especially in chronic kidney disease (CKD). The elevation of them in CKD might cause confusion and misuse of these tumor markers. We conducted this retrospective study to investigate which of the five widely used tumor markers including carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), cytokeratin 19 fragment antigen 21-1 (Cyfra21-1), squamous cell carcinoma antigen (SCC) and neuron specific enolase (NSE) are affected markedly by CKD, in order to use them more effectively.</p><p><b>METHODS</b>Serum tumor marker concentrations, biochemical, hematological parameters, and urinalysis were measured in CKD patients and healthy controls. The positive rate and median tumor markers' level in CKD patients and controls, and those in CKD patients stratified by CKD grade were compared using nonparametric rank tests. Correlation analysis of serum tumor markers and other parameters in CKD patients were performed using the Spearman correlation coefficient. Multivariate Logistic regression analysis was used to estimate the important variables that caused elevated serum concentrations of these markers in CKD patients.</p><p><b>RESULTS</b>The overall positive rates and serum concentrations of Cyfra21-1, SCC, CEA in CKD group were significantly higher than those in control group. Positive rate and serum concentrations of those tumor markers increased as kidney function decreased. Both univariate analysis and multivariate regression analysis showed that the elevations of those tumor markers were not only associated with kidney function, but also with nutritional status.</p><p><b>CONCLUSIONS</b>Serum concentrations of Cyfra21-1, SCC, CEA are significantly influenced by kidney function, as well as nutritional status. Therefore, in clinical work, the indices of kidney function and nutritional status could be simultaneously measured to improve interpretation of the results of those tumor marker concentrations.</p>

Clinical observation of jianpi bushen formula to prevent collapse of osteonecrosis of femoral head / 中国中药杂志

To observe the clinical effect of Jianpi Bushen formula to prevent the collapse of osteonecrosis of femoral head (ONFH) of type phlegm and blood stasis obstructing the collaterals. 50 cases (including 73 hips) of non-collapse ONFH (ARCO I, II, III a) were selected from the out-patient department of orthopedic in Guang'anmen Hospital attached to China Academy of Chinese Medical Science. All the cases fit for diagnostic criteria were given Jianpi Bushen formula and followed up. Staging criteria was ARCO classification. Harris score evaluated the hip function. The mean follow-up time was 4.2 years (3-5.4 years). After a mean of 4.2 years (3-5.4 years) followed-up, 12 hips collapse occurred while 61 hips not (the non-collapse rate was 83.56%). Collapse did not occurred among the all 7 hips at stage ARCO I (the non-collapse rate was 100%). Among the 49 hips at stage ARCO II, 9 hips collapse occurred while 40 hips not (the non-collapse rate was 81.63%). Among the 17 hips at stage ARCO III a, 3 hips collapse occurred while 14 hips not (the non-collapse rate was 82.35%). Kaplan-Meier analysis showed the average survival time of non-collapse was 5 years (4. 8-5.2 years). The 3 year survival rate of non-collapse was 92.5%. The 4 year survival rate was 74% and the condition tended to be stability trend. The Harris score was 71.93 +/- 11.25 before treatment and 81.63 +/- 12.16 after treatment, significantly different. These results suggest that: Jianpi Bushen formula is an effective method for treating ONFH of type phlegm and blood stasis obstructing the collaterals. It can delay or prevent the collapse of femoral head and significantly improve the hip function.

Effect of removable partial dentures restoration on oral health-related quality of life of patients with shortened dental arch / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To assess the effect of removable partial dentures (PRD) restoration on oral health-related quality of life (OHRQOL) of subjects with shortened dental arches (SDA) using the Chinese version of the Oral Health Impact Profile (OHIP-14).</p><p><b>METHODS</b>Consecutive patients with shortened dental arches were recruited from the Department of Prosthodontics, Foshan Chancheng Hospital of Stomatology. The Chinese version of OHIP-14 was administered to each subject before treatment and after treatment. The subjective outcomes of removable partial dentures therapy on SDA were collected and the pre- and post-treatment scores of the Chinese version of OHIP-14 were compared.</p><p><b>RESULTS</b>After treatment, significant decrease in patients' total score of OHRQOL [before: 9 (6, 12) ; after: 4 (2.25, 6)] was detected, as well as physical pain subscale, physical disability subscale and handicap subscale; and the score of OHRQOL in function limitation subscale increased [before: 0(0,0); after: 2(1, 2)].</p><p><b>CONCLUSIONS</b>The main impacts of shortened dental arch with intact anterior region affecting patients' OHRQOL are physical disability.From a quality-of-life perspective, patients with SDA can perceive benefits from RPD.</p>

Influence of Shenqing Recipe on morphology and quantity of colonic interstitial cells of Cajal in trinitrobenzene sulfonic acid induced rat colitis / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To observe the influence of Shenqing Recipe (SQR), a kind of Traditional Chinese Medicine, on the morphology and quantity of colonic interstitial cells of Cajal (ICC) in trinitrobenzene sulfonic acid (TNBS)-induced rat colitis, and to investigate the possible mechanism of SQR in regulating intestinal dynamics.</p><p><b>METHODS</b>Sixty rats were randomly divided into normal control, model 1, model 2, mesalazine, and high-dose, and low-dose SQR groups with 10 rats in each group. TNBS (10 mg) dissolved in 50% ethanol was instilled into the lumen of the rat colon of the latter five groups to induce colitis. On the 4th day after administration of TNBS, each treatment group was administered one of the following formulations by enteroclysis gavage once a day for 7 days: 600 mg•kg⁻¹•d⁻¹ mesalazine, 2.4 g•kg⁻¹•d⁻¹ SQR, and 1.2 g•kg⁻¹•d⁻¹ SQR. Model 2 rats received normal saline solution. After 7 days colonic samples were collected. While the colonic samples of model 1 group were collected on the 3rd day after TNBS administered. Ultrastructure of ICC in the damaged colonic tissues was observed with transmission electron microscope. Expression of c-kit protein in colonic tissue was determined by immunohistochemical staining and Western blot.</p><p><b>RESULTS</b>The ultrastructure of colonic ICC in the rat model of TNBS-induced colitis showed a severe injury, and administration of SQR or mesalazine reduced the severity of injury. Similarly, the expression of c-kit protein of TNBS-induced colitis rat model was significantly decreased compared with the normal control group (P < 0.05). Treatment with SQR or mesalazine significantly increased the expression of c-kit protein compared with the administration of control formulations (P < 0.05), especially the high-dose SQR group.</p><p><b>CONCLUSION</b>SQR could alleviate and repair the injured ICC, and improve its quantity, which might be involved in regulating intestinal motility.</p>

Feasibility of peptide mass fingerprinting for differential diagnosis of IgA and non-IgA nephropathy / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the feasibility of peptide mass fingerprinting for non-invasive differential diagnosis of IgA nephropathy (IgAN) from the non-IgA nephropathy (IgAN).?</p><p><b>METHODS</b>According to the results of renal biopsy, 56 patients were divided into IgAN group (n=28) and non-IgAN group (n=28), and peptide mass fingerprints were acquired from these patients using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).</p><p><b>RESULTS</b>Nine different peptides were identified between IgAN and non-IgAN. The two most distinctive differentially expressed peptides, with peptide peak values of 4476.46 and 1968.10, showed area under curve values of 86.18% and 79.77%. Principal component analysis demonstrated that the accumulated explained variance of the first 8 differential peptides reached 95%, suggesting the feasibility of differential diagnosis of IgAN from non-IgAN. Comparison with the Matrix protein database identified the peptide with a relative molecular mass of 5338.08 as a fragment of mucin 4 inform and the 2082.77 peptide as fragment of α1-II type collagen inform.</p><p><b>CONCLUSION</b>MALDI-TOF MS is feasible for differential diagnosis of IgAN and non-IgAN and also has great potentials in the classification of the subtypes of other systemic diseases.</p>

Effect of small interfering RNA targeting Rac1 gene on colony formation of SW480 cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a vector expressing small interfering RNA (siRNA) against Rac1 gene and observe its effect on soft agar colony formation of SW480 cells in vitro.</p><p><b>METHODS</b>Oligos of 64 base pairs for hairpin RNA targeting Rac1 were chemically synthesized and annealed. The siRNA constructs for Rac1, produced by inserting the annealed oligos into the downstream of H1 promoter of linearized pSUPER, were confirmed by restriction digestion and DNA sequencing. The constructed Rac1-siRNA was transfected into SW480 cells and Western blotting was performed to assess the expression and interference efficiency of siRNAs against Rac1.The soft agar colony formation assay was used to study the effect of Rac1 gene silencing on SW480 cells.</p><p><b>RESULTS</b>Restriction digestion and DNA sequencing showed that the siRNA targeting Rac1 gene was successfully constructed. The siRNA could effectively down-regulate the expression of Rac1 in SW480 cells. Soft agar colony formation assay showed that the colony number and diameter of SW480 cells was reduced after siRNA transfection.</p><p><b>CONCLUSION</b>A vector expressing hairpin RNA against Rac1 gene are successfully produced, which significantly reduces the colony numbers and size of SW480 cells in vitro, suggesting that Rac1 plays an important role in the growth of colorectal cancer in vitro.</p>

The study on metabolic difference of human body affected by active stress and passive stress under special events / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the metabolic difference of body influenced by active stress and passive stress under special events.</p><p><b>METHODS</b>To detect serum multiple biochemistry index of 57 earthquake rescue medical team and 13 victims of a natural calamity in Wenchuan earthquake by using Hitachi 7600 automatic analyzer.</p><p><b>RESULTS</b>Stress affected biochemistry index deeply. To compared with rescue medical team, the serum ADA, ALP and TG of victims increased obviously and TP, ALB, MAO, Cr, UA, K, Na, Cl, Ca, ApoA1 and HDL decreased obviously.</p><p><b>CONCLUSION</b>Many biochemistry index have been changed under stress and it relate with stress extent. The human body function status was better in active stress than in passive stress.</p>

Effect of p21-activated kinase-1 on the invasiveness of colorectal carcinoma cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of p21-activated kinase-1 (PAK1) gene transfection on the invasiveness of human colorectal carcinoma SW480 cells in vitro.</p><p><b>METHODS</b>SW480 cells in routine culture were transfected with the recombinant plasmid EGFP-C1/PAK1 via Lipofectamine(TM) 2000. The expression of PAK1 protein in SW480 cells was detected using Western blotting, and the changes of the invasiveness of SW480 cells were evaluated using Boyden chamber invasion assay.</p><p><b>RESULTS</b>Forty-eight hours after transfection with pEGFP-C1/ PAK1, the PAK1 protein expression increased significantly in comparison with those in negative and vector control groups. The invasiveness of the SW480 cells was significantly enhanced after the transfection.</p><p><b>CONCLUSION</b>The PAK1 gene transfection can increase the expression of PAK1 in SW480 cells and enhance the invasiveness of the cells. PAK1 can be associated with the invasiveness and metastasis of colorectal carcinoma cells.</p>

Effect of X-ray exposure on soluble tumor necrosis factor receptor-p75 release in hepatocellular carcinoma HepG2 cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of X-ray exposure on the release of soluble tumor necrosis factor receptor-p75 (sTNFR-p75) in hepatocellular carcinoma HepG2 cells in vitro.</p><p><b>METHODS</b>Enzyme-linked immunosorbent assay (ELISA) was used to examine the levels of sTNFR-p75 in the supernatants of HepG2 cells before and after X-ray exposure. The cell apoptosis was analyzed by flow cytometry and transmission electron microscope(TEM), and the morphological changes of the cells were examined under optical microscope and transmission electron microscope(TEM).</p><p><b>RESULTS</b>X-ray exposure of the cells resulted in a strong increase of cell apoptosis (P<0.05) and sTNFR-p75 production in the cells as compared with the those before the exposure (P<0.01). Optical microscopy revealed apoptotic changes of HepG2 cell after the exposure, shown as cell shrinkage, spherical cell morphology, cytoplasmic and nuclear condensation. Apoptotic bodies were detected by TEM.</p><p><b>CONCLUSION</b>X-ray exposure induces HepG2 cells apoptosis by inhibiting the release of sTNFR-p75 into the supernatant.</p>