Effect of Ursolic Acid on Proliferation and Apoptosis of Colorectal Cancer Cells Based on Akt/FoxO Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effects and molecular mechanism of ursolic acid on the proliferation and apoptosis of colorectal cancer cells. MethodThe proliferation inhibition rate of human colorectal cancer RKO cells treated with different concentrations of ursolic acid （0， 5， 10， 15， 20， 25， 30 μmol·L-1） was detected by cell counting kit-8 （CCK-8）， and the half maximal inhibitory concentration （IC50） at 24 h and 48 h was calculated. According to the IC50 of RKO cells treated with ursolic acid for 24 h， two concentrations were selected for subsequent experiments. The colony formation assay was used to detect the proliferation ability of the cells and flow cytometry was used to detect the apoptosis rate and cell cycle arrest after treatment of RKO cells with ursolic acid. After treatment of RKO cells with ursolic acid for 24 hours， the expression of B-cell lymphoma 2 （Bcl-2）-associated X protein （Bax） in RKO cells， Bcl-2 in Raji cells， PMA responsive gene in T lymphocyte （Noxa）， cyclin-dependent kinase inhibitor 1A （p21）， cyclin-dependent kinase inhibitor 1B （p27）， cyclin-dependent kinase 4 （CDK4）， protein kinase B （Akt）， phosphorylated Akt （p-Akt）， forkhead transcription factor O3a （FoxO3a）， and phosphorylated FoxO3a （p-FoxO3a） was determined by Western blot. ResultCompared with the blank group， the ursolic acid groups could inhibit the viability of RKO cells （P<0.05， P<0.01）， and the colony formation rates of RKO cells in the ursolic acid groups were reduced （P<0.05， P<0.01） in a concentration-dependent manner. The cells in the ursolic acid group （20 μmol·L-1） experienced cell cycle arrest， which increased in the early stage of synthesis， ie， the G0/G1 phase （P<0.05） as compared with the results in the blank group. Compared with the blank group， the ursolic acid groups （15 and 20 μmol·L-1） showed increased protein expression of p21 and p27， decreased expression of CDK4 protein （P<0.05， P<0.01）， and increased apoptosis rate， and the ursolic acid group （20 μmol·L-1） showed increased protein expression of Bax and Noxa and decreased expression of Bcl-2 （P<0.05， P<0.01）. In terms of mechanism， compared with the blank group， the ursolic acid group （20 μmol·L-1） down-regulated the expression of p-Akt protein and up-regulated the expression of p-FoxO3a （P<0.05， P<0.01）， and there was no significant change in the total protein of Akt and FoxO3a. ConclusionUrsolic acid can effectively inhibit the proliferation of colorectal cancer RKO cells and promote cell apoptosis， which may be related to the Akt/FoxO pathway.

Application of Self-assembled Nano-strategies of Traditional Chinese Medicine in Tumor Therapy： A Review / 中国实验方剂学杂志

Chinese medicine self-assembly nano-strategies（CSAN） is to utilize the self-assembly property of Chinese medicine components， so that the Chinese medicine components can self-assemble to form structurally stable nano-preparations through non-covalent interactions. The formation of Chinese medicine self-assembly nano-preparations is often a synergistic result of a variety of non-covalent interactions， and many Chinese medicine monomers are susceptible to self-assembly due to their structural characteristics， and the phenomenon of self-assembly of Chinese medicine is also common in the decoction of single or compound Chinese medicine， which has attracted the attention of researchers. It is found that CSAN can improve the solubility and bioavailability of active components in Chinese medicine， which is of positive significance for the development and application of insoluble components of Chinese medicine. The self-assembly phenomenon of Chinese medicine decoction is closely related to the therapeutic efficacy， and the study of self-assembly phenomenon of Chinese medicine will bring a new perspective for the explanation of the mechanism of Chinese medicine decoction. At the same time， traditional Chinese medicine（TCM） has unique advantages in the field of anti-tumor. The application of CSAN in the field of oncology can not only exert the anti-tumor effect of the active components of Chinese medicine directly， but also act as a natural nano-carrier to carry chemotherapy drugs for combination chemotherapy， improve the targeting of drugs， enhance the anti-tumor efficacy， and reduce the side effects of chemotherapy， which has excellent anti-tumor potential. The preparation method of Chinese medicine self-assembly nano-preparations is simple， low cost， and has better safety than traditional nano-preparations， which is conducive to the promotion of the clinical transformation of nano-preparations， and also helps to provide new strategies and perspectives for promoting the modernization of TCM. Therefore， based on a large number of researches in this field in recent years， this paper reviewed the formation mechanism， different assembly forms， formation conditions and stability of Chinese medicine self-assembly nano-preparations by searching databases such as China national knowledge infrastructure（CNKI）， PubMed， WanFang data and VIP， and summarized the application of CSAN in different tumor therapies， providing a reference for further research on CSAN.

Value of central vein sign and iron deposition to differentiate multiple sclerosis from neuromyelitis optica spectrum disorder / 中华放射学杂志

Objective:To investigate the value of central vein sign (CVS) and iron deposition on quantitative susceptibility imaging (QSM) of 3.0 T MRI in differentiating multiple sclerosis (MS) from neuromyelitis optica spectrum disease (NMOSD).Methods:This study was a retrospective study. A total of 54 MS patients and 49 NMOSD patients were enrolled from July 2018 to December 2020 in People′s Hospital of Leshan and the First Affiliated Hospital of Chongqing Medical University. All patients underwent conventional MRI and three-dimensional enhanced T 2*-weighted angiography (3D-ESWAN), and ESWAN-filtered phase and QSM were reconstructed from 3D-ESWAN data. First, brain lesions of MS and NMOSD were screened on proton density (PD)-T 2WI, and then the location of lesions, CVS and nodular/annular iron deposition were observed on phase and QSM images. The χ 2 test was used to compare the differences in intracranial lesion location, CVS and iron deposition between MS and NMOSD patients. Receiver operating characteristic curve and area under the curve (AUC) were used to assess the efficiency of CVS and QSM iron deposition to differentiate MS from NMOSD. Results:A total of 968 MS lesions were observed in 54 MS patients, of which CVSs were found in 354 lesions and 227 CVSs were located around the lateral ventricles, 117 in deep white matter (DWM) and 10 in the cortex/subcortex; 372 lesions showed nodular iron deposition, and 193 lesions ring iron deposition on QSM. Totally 247 brain lesions were observed in 41 of 48 patients with NMOSD, of which CVSs were found in 4 lesions and 1 located around the lateral ventricle, 3 located in the DWM; 3 lesions showed nodular iron deposition on QSM. There were significant differences in cortex/subcortex lesions, CVS and iron deposition between MS and NMOSD patients (χ 2 were 29.33, 115.66 and 258.21, respectively, all P<0.001). The AUC of CVS for differentiating MS from NMOSD was 0.941 (95%CI 0.887-0.994), with a sensitivity of 96.3% and a specificity of 91.8%; the AUC of iron deposition for differentiating MS from NMOSD was 0.969 (95%CI 0.930-1.000), with a sensitivity of 100% and a specificity of 93.9%. Conclusion:CVS and iron deposition on 3.0 T MRI are distinct radiologic features of MS lesions from those of NMOSD lesions, and have certain value in the differential diagnosis.

Research Development in Patients with Multiple Sclerosis by Focus on MRI / 罕见病研究

Multiple sclerosis (MS) is an autoimmune inflammatory demyelinating disease of the central nervous system whose clinical manifestations and diagnostic criteria are non-specific and absence of golden criteria when diagnosing. Magnetic resonance imaging (MRI) can not only observe the focal lesions of demyelination, but also evaluate microstructural damages and iron deposition in the white/gray matter in MS patients by applying various developing sequences. MRIs play an irreplaceable role in revealing pathological evolution, prognosis and monitoring after treatment of MS patients. This paper reviewed the clinical value and application of MRIs for the MS patients in recent years.

Neonatal Ebstein's anomaly with type B Wolff-Parkinson-White syndrome: a case report / 中华围产医学杂志

This paper reported a male newborn with Ebstein's anomaly complicated by type B Wolff-Parkinson-White syndrome. The baby was delivered at 39 weeks of gestation and diagnosed as Ebstein's anomaly complicated by type B Wolff-Parkinson-White syndrome 2 d after birth. He was improved and discharged after conservative management in the Department of Internal Medicine. No abnormality was found by fetal echocardiography at 20 weeks of gestation, while supraventricular tachycardia was diagnosed by M-mode ultrasonography at 38 weeks of gestation due to rapid fetal heart rate. In addition to hemodynamic changes,the abnormal cardiac structure in patients with Ebstein's anomaly may also lead to tachyarrhythmia. Therefore, in fetuses diagnosed with supraventricular tachycardia by prenatal ultrasound or children with type B Wolff-Parkinson-White syndrome detected by electrocardiogram after birth, tricuspid valve should be carefully scanned under echocardiography to avoid the missed diagnosis of Ebstein's anomaly.

Effects of femtosecond laser treatment on surface characteristics and flexural strength of zirconia / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the effects of femtosecond laser treated microgrooved surface on microscopic topography, phase transformation, and three-points flexural strength of zirconia, and to provide reference for surface microstructure optimization of zirconia implant.@*METHODS@#According to different surface treatment methods, 57 computer aided design/computer aided manufacture (CAD/CAM) zirconia bars (20.0 mm×4.0 mm×1.4 mm) were evenly divided into three groups: sintered group, no treatment after sintering, taken as control; sandblasted group, sandblasted with 110 μm aluminium oxide (Al2O3) after sintering; microgrooved group, femtosecond laser fabricated microgrooves with 50 μm width, 30 μm depth, and 100 μm pitch. Surface microscopic topography was observed with scanning electron microscope (SEM) and 3D laser microscope. Further, surface roughness in each group and microgroove size were measured. Crystal phase was analyzed with X-ray diffraction. Specimens were subjected to three- points flexural strength test, and Weibull distribution was used to analyze their strength characteristics.@*RESULTS@#SEM showed that sintered surface was flat with clear grain structure; sandblasted surface exihibited bumps and holes with sharp margins and irregular shape; microgrooves were regularly aligned without evident defect, and nano-scale particles were observed on the surface inside of the microgrooves. Ra value of microgrooved group [(9.42±0.28)] μm was significantly higher than that of sandblasted group [(1.04±0.03) μm] and sintered group [(0.60±0.04) μm], and there was statistical difference between sandblasted group and sintered group (P < 0.001). The microgroove size was precise with (49.75±1.24) μm width, (30.85±1.02) μm depth, and (100.58±1.94) μm pitch. Crystal phase analysis showed that monoclinic volume fraction of sandblasted group (18.17%) was much higher than that of sintered group (1.55%), while microgrooved group (2.21%) was similar with sintered group. The flexural strength of sandblasted group (986.22±163.25) MPa had no statistical difference with that of sintered group (946.46±134.15) MPa (P=0.847), but the strength in microgrooved group (547.92±30.89) MPa dropped significantly compared with the other two groups (P < 0.001). Weibull modulus of sintered, sandblasted, microgrooved groups were 7.89, 6.98, and 23.46, respectively.@*CONCLUSION@#Femtosecond laser was able to form micro/nanostructured microgrooves on zirconia surface, which deleteriously affected the flexural strength of zirconia.

Advances in the application of confocal microscopy to keratopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#Confocal microscopy serving as an <i>in vivo</i> histological examination for cornea holds superiority in disease diagnosis, assessment and follow-up as well as physiological and pathological study due to it's real-time, non-invasive, quick operation with high-resolution imaging, and has been widely applied in ophthalmic clinic and research. This article reviewed the recent advances in the application of confocal microscopy in infectious or degenerative corneal diseases, and ocular surface diseases caused by systemic diseases.

Promoting effect of 810 nm low-level laser on axonal regeneration of neurons in mice with spinal cord injury and its related mechanism / 中华创伤杂志

Objective To investigate the effect of 810 nm low-level laser on neuronal axonal regeneration of mice with spinal cord injury and its related mechanism.Methods In vivo experiment:20 Balb/c mice were randomly divided into the spinal cord injury group(SCI group)and the 810 nm low-level laser irradiation group(low-level laser group)after spinal cord injury according to the random number table method,with each group containing ten mice.A mice SCI model was established through clamp injury and the low-level laser group continuously irradiated the damaged area with weak 810 nm low-level laser with selected parameters(continuous wave with wave length 810 nm,power density 2 mW/cm2,spot are 4.5 cm2,irradiation time 50 minutes,energy 6000J/cm2).Then immunofluorescence staining was used to observe the M1 macrophage marker-inducible nitric oxide synthase(iNOS),the M2 macrophage marker arginase 1(Arg-1)and the universal marker F4/80 of macrophages after 14 days.Furthermore,in the in vitro experiment,standardized low-level laser-macrophage irradiation model was established.Another 20 Balb/c mice were used to obtain primary bone marrow-derived macrophages which were induced into M1 macrophages using lipopolysaccharide(LPS)and interferon-gamma(INF-γ).The M1 macrophages were randomly divided into the M1 macrophage group(M1 group)and the low-level laser therapy group(M1 + low-level laser group)equally according to the random number table method.The M1 group was not treated,and the M1 + low-level laser group was treated with low-level laser of selected parameters.RT-qPCR and ELISA were used to detect the expression of interleukin-1 receptor antagonist(IL-1RA)and interleukin-10(IL-10)in M1 macrophages 24 hours after irradiation.Western blot was used to analyze the expression of iNOS,Arg-1,differentiation antigen cluster 206(CD206),protein kinase B(AKT),phosphorylated protein kinase B(p-AKT),cyclic adenosine response element binding protein(CREB)and phosphorylated cyclic adenosine response element binding protein(p-CREB)in M1 macrophages 48 hours after irradiation.Dorsal root gangtion neurons(DRG)were cultured in two groups of macrophage conditioned medium,and the length of DRG axon growth was measured 48 h later to evaluate the effect of low-level laser on neuronal axon growth.Results In the in vivo experiment,compared with mice with spinal cord injury alone,the fluorescence intensity of F4/80+ iNOS+ in the spinal cord injury area decreased(1.00±0.08vs. 0.06±0.04)(P＜ 0.05)and the fluorescence intensity of F4/80 + Arg-1 + increased after low-level laser(1.00±0.07vs.2.15±0.12)(P＜0.01).In the in vitro experiment,compared with the M1 group,the expression of the M1 macrophage marker iNOS in the M1 + low-level laser group decreased(1.00±0.11 vs.0.08±0.01)(P＜ 0.01);the M2 macrophage marker Arg-1(1.00±0.14vs.2.44±0.16)(P＜0.01),and the expression of CD206(1.00±0.12 vs.1.83±0.05)(P＜0.01)increased.In addition,IL-1RA expression was increased in the M1 + low-level laser group compared with the M1 group(RT-qPCR:1.00±0.00vs.2.27±0.22)(P＜0.01)(ELISA:1435.58±100.48vs.2006.12±123.91(P＜0.05);IL-10 expression was also increased in the M1 +low-level laser group compared with the M1 group(RT-qPCR:1.00±0.00 vs. 3.45±0,56)(P＜0.05)(ELISA:137.13±4.20 vs.188.29±8.49)(P＜ 0,01);compared with the M1 group,the macrophage polarization pathway protein in the M1 + low-level laser group increased,AKT(1.07±0.12vs.1.74±0.04)(P＜0.01),p-AKT(1.00±0.12 vs.1.64±0.15)(P＜0.05),p-CREB(1.00±0.10vs.2.12±0.18)(P＜0.01).Compared with the M1 group,the conditioned medium of the M1 + low-level laser group significantly promoted DRG axon growth(567.66±63.59 vs.1068.95±130.14)(P＜ 0,05).Conclusions The 810 nm low-level laser irradiation can promote neuronal axon regeneration of mice with spinal cord injury,which may be related to the regulation of macrophage polarization phenotype by low-level laser through AKT/CREB pathway.

Preparation of sorafenib and doxorubicin loaded nanodroplets and its ultrasound-sensitive in vitro:experimental study / 中华超声影像学杂志

Objective To prepare ultrasound(US)responsive nanodroplets(NDs)simultaneously loaded with anticancer drug Sorafenib(SF)and Doxorubicin(DOX),and to characterize its ultrasound responsibility in vitro and in vivo.Methods The SF/DOX NDs were prepared using the thin-film hydration method.The particle diameter,Zeta potential and drug-encapsulation efficiency were characterized.The acoustic droplets vaporization activity was monitored by in vitro ultrasound imaging and light microscope. The cavitation effect was monitored by in vitro ultrasound imaging and transmission electron microscopy. Results SF/DOX NDs were round in shape,the mean diameter and Zeta potential of SF/DOX NDs was (498 ± 67.34)nm,-(38.87 ± 3.78)mV,respectively.The entrapment efficiency of SF and DOX was (58.14±2.93)%,(51.23±4.11)%,respectively.SF/DOX NDs underwent a phase transition into bubbles and could be continuously imaged for more than 25 min in vitro,and afterward therapeutic ultrasound pulse induced inertial cavitation and substantially enhanced treatment.Conclusions US-responsible SF/DOX NDs are prepared using thin-film hydration mehtod,it can enhance ultrasonic echo in vitro and release anticancer drug by the aid of US exposure,which possesses greater researching and applicating value.

Clinical effects of internal fixation with Herbert screws for the treatment of Pipkin femoral head fractures / 中国骨伤

<p><b>OBJECTIVE</b>To explore the therapeutic effects of internal fixation with Herbert screws for the treatment of Pipkin type I and type II femoral head fractures.</p><p><b>METHODS</b>From January 2008 to December 2012, 23 patients with Pipkin type I and type II femoral head fractures were treated with open reduction and internal fixation by Herbert screws through Kocher-Langenbeck approach. Twenty-three patients (aged 18 to 52 years with a mean of 35.5 years, including 18 males and 5 females patients, 8 left hips and 15 right hips) with femoral head fractures and posterior hip dislocation. The fracture was classified according to Pipkin classification based on the radiographic findings, 5 patients had type I and 18 had type II fractures. The duration time from admission to the operation ranged from 6 to 72 h (averaged 32 h). The clinical and radiographic outcomes of the patients were measured using Thompson-Epstein scoring scale. The Harris hip score(HHS) was used to evaluate and compare hip functions at the latest follow-up between affected and healthy sides.</p><p><b>RESULTS</b>All the patients were followed up, and the duration ranged from 20 to 48 months (averaged 30 months). According to Thompson-Epstein system, 12 patients got an excellent result, 6 good, 4 fair and 1 poor. The average HHS at the finial follow-up was 87.80 ± 8.46 (ranged from 66 to 95), which is similar to that in the healthy side 90.10 ± 6.35 (ranged from 72 to 98) (t = 1.044, P = 0.302). The complications such as deep infection, and deep vein thrombosis were not found. At the 3rd year during follow-up,4 patients had avascular necrosis of femoral head. At the 1st year of follow-up, 1 patient had hip pain after walking,screws loosening and shift after trauma,and serious complications of traumatic arthritis. All the 5 patients were treated with total hip arthroplasties.</p><p><b>CONCLUSION</b>The treatment of internal fixation with Herbert screws through the Kocher-Langenbeck approach is effective for Pipkin type I and type II femoral head fracture. The method is reliable and valuable for recommendation. However, such fracture may have avascular necrosis of femoral head and complication of traumatic arthritis, which should be observed carefully in clinic with preparation of the prevention and treatment measures.</p>

The effect of Shugan-Yishen formula on HER2-ERK/MAPK-ERα Pathway of TAM-resistant breast cancer cell lines / 国际中医中药杂志

Objective To investigate the effect of Shugan-Yishen formula on Tamoxifen-resistant MCF-7 cell line(LCC9) through HER2-ERK/MAPK-ERα pathway. Methods Four serums were prepared with the method of serum pharmacology:the serum of Shugan-Yishen formula, Shugan-Yishen formula plus Tamoxifen(TAM), blank and TAM alone. After LCC-9 cells were affected in the above-mentioed serums in vitro, the inhibition rate and the level of mRNA as well as the protein expression of HER2, ERα, ERK/MAPK, p-ERK/MAPK were detected. Results MTT showed that TAM alone group、Shugan-Yishen formula group and the Shugan-Yishen formula plus TAM group had higher inhibition rates at 48th hour, the inhibition rates was(6.61±0.129)%, (47.43±2.24)%, (54.19±3.364)%, compared to the blank group, Shugan-Yishen formula group and the Shugan-Yishen formula plus TAM group had a stronger inhibition on LCC9 at 48th hour (P0.05). RT-PCR result showed that compared to the blank group, the expression levels of ESR1 mRNA of group of Shugan Yishen formula plus TAM werehigher by and large at 48th hour (P0.05). Conclusions ①The combination of Shugan-Yishen formula with tamoxifen can inhibit the LCC9 cells. The inhibition effect reaches the peak at 48th hour. ②The mechanism of Shugan-Yishen formula plus TAM combination effects on human breast cancer TAM-resistant cell line LCC9 may improve the sensitivity of ERαby inhibiting the expression of HER2, ERK/MAPK.

Research on cultivation for comprehensive humanistic quality of oral medical students, which is oriented by competency / 华西口腔医学杂志

Medicine has dual features of humanities and natural science. Thus, it is necessary for the development of modern higher education to carry out the humanistic quality education to medical students. It is not only the request of modern medical model and medical development, but also the urgent need of the development of medical and health. Besides, it plays an important part in the cultivation of medical students. In the face of the urgent need of cultivation for comprehensive humanistic quality of oral medical students, West China School of Stomatology, Sichuan University, led in the cultivation method which is oriented by competency, give some suggestions to deal with students' lack of language ability, humanistic concern to patients and aesthetic accomplishment. And it has already achieved a better teaching effect.

Competency oriented management: a training pattern of elite stomatology student / 华西口腔医学杂志

The competency oriented management theory and method, which was proved to be an effective management measure, have been put into practice in many areas including high education. This essay will probe into the possibility of introducing the competency oriented management theory into the education of undergraduate stomatology students, analyze and research its feasibility from a theoretical point of view by combing the features of stomatology students and the patterns of educating stomatology students. This essay proposes a new pattern of training elite students of stomatology major with competency as the orientation and summarizes the practical experiences in training such elite students.

Treatment of fracture of fifth metatarsal base and degree III lateral ligament injury of ankle with compressed cannulated screw and suture anchors / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the clinical outcomes of surgical treatment of fracture of the fifth metatarsal base combined with degree III lateral ligament injury of ankle.</p><p><b>METHODS</b>From January 2008 to December 2011, 32 patients with fracture of the fifth metatarsal base combined with degree III lateral ligament injury of ankle were treated with surgery. Fractures were fixed with compressed canulated screw and ligaments were repaired with suture anchors. After operation, ankle joints were fixed in neutral position and slightly valgus position by plaster slab. Taking out stitch was performed at 2 weeks after operation and non-weight loading walking by double crutches support started; after the 6 weeks, remove the gypsum and part-weight loading walking by brace protection; at the 8 weeks after operation, completely weight loading walking was permitted. American Orthopaedic Foot and Ankle Society (AOFAS) was used to evaluate the clinical effect.</p><p><b>RESULTS</b>Thirty-two patients were followed up from 8 to 18 months with an average of 12 months. All fractures obtained healing with an average time of 12.5 weeks (ranged, 8 to 24 weeks). According to the standard of AOFAS, 18 cases got excellent results and 14 good.</p><p><b>CONCLUSION</b>The method that fracture fixation with compressed canulated screw and ligament repair with suture anchors can obtain satisfactory effects in treating fracture of the fifth metatarsal base and degree III lateral ligament injury of ankle.</p>

Detection of WU polyomavirus in children with low respiratory tract infections using real-time fluorescent quantitative PCR / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Development and application of a real time fluorescent quantitative PCR (FQ-PCR) assay for detecting WU polyomavirus in children with low respiratory tract infections.</p><p><b>METHODS</b>The VP2 gene of WU polyomavirus was selected as the detection target, from which the real time primers and probes were designed. The standard curve was established by using recombinant plasmid as template. And the FQ-PCR assay for specific detection of WU polyomavirus was established. The specificity, sensitivity and reproducibility of the method were evaluated. Furthermore, the clinical specimens from children with respiratory tract infections collected in Wenling First People's Hospital were quantitatively detected using this method.</p><p><b>RESULTS</b>In this study, the FQ-PCR method was established to detect a specific fragment in VP2gene of WU polyomavirus. The standard curve coefficient R2 was 0.998. And this method can detect as low as 50 copies recombinant plasmid. The clinical specimens of sputum and throat swab from children with respiratory tract infections were quantitatively detected using this method. 7 sputum specimens were detected as WU polyomavirus positive in 700 sputum specimens, the positive ratio was 1.00%. No positive specimens were detected in 146 specimens of throat swabs and 846 blood samples from same patient population.</p><p><b>CONCLUSION</b>The results indicated that the FQ-PCR assay method established in this study was specific, rapid and sensitive for detecting WU polyomavirus in children with lower respiratory tract infections. The sputum specimen is more suitable to be used for gene detection of WU polyomavirus than throat swab or blood.</p>

Detection of BK virus infection in renal transplant recipients and clinical application / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the detection methods of BK virus infection in kidney transplant recipients, and to explore the clinical application.</p><p><b>METHODS</b>132 cases of renal transplant recipients were undertaken BK virus detection including presence of decoy cells in urinary sediment, urine and serum BKV-DNA to demonstrate the BK virus replication.</p><p><b>RESULT</b>Among 132 cases of renal transplant recipients, urinary decoy cell was found in 37 (28.0%) patients and the median time was 12 months after surgery. 32 (24.2%) patients were diagnosed as BK viruria at a median of 11 months after surgery, and 16 (12.1%) recipients were diagnosed as BK viremia at a median of 15 months after surgery, 5 patients with BK viruria were diagnosed as BK virus associated nephropathy according to allograft biopsy.</p><p><b>CONCLUSION</b>To make early diagnosis of BK virus infection, detection of urine decoy cells and BKV-DNA in urine and plasma sample is important,which provides an important basis for the prevention of BK virus associated nephropathy.</p>

Expression of MCP-1 in the pancreas of piglet with chronic obstructive pancreatitis and its significance / 中华胰腺病杂志

Objective To determine the serum concentration of MCP-1 and the expression of MCP-1 protein in the pancreas in the piglet with chronic obstructive pancreatitis and to explore the role of MCP-1 protein in pancreatic fibrosisits.Methods The piglet model of chronic obstructive pancreatitis was established by incomplete ligation of the pancreatic duct.The piglets were sacrificed at 4, 6, 8 weeks after induction.Pathological changes of pancreas were examined.Pancreatic fibrosis was assessed by VG staining.Serum MCP-1 concentrations were detected by ELISA method.MCP-1 and α-SMA, PDGF, TGF-β1 and NF-κB protein expression were detected by immunohistochemistry.Results The induction was successful in 14 piglets ( 58.3% ).Mild atrophic changes, interstitial fibrosis, chronic inflammatory cell infiltration could be observed in the body and tail of pancreas from the 4th week in the experimental group.The most obvious changes occurred in the 8th week.Stage Ⅰ pancreatic fibrosis occurred in 5 piglets (35.7%), stage Ⅱ in 4 piglets (28.6%), stage Ⅲ in 5 rats ( 35.7% ).Seurm MCP-1 at 4, 6, 8 weeks was ( 102.44 ± 36.25 ) pg/ml,(97.84 ± 28.67) pg/ml, ( 94.32 ± 28.42 ) pg/ml, respectively, and was significantly higher than that in control group [ ( 10.42 ±5.86) pg/ml, (8.58 ±4.86) pg/ml, (8.22 ±4.58) pg/ml, P ＜0.01 ].There was no MCP-1 protein expression in the control group;MCP-1 protein was detected in the successful induction group, and MCP-1 expression was positively correlated with expressions of the PDGF, TGF-β1, α-SMA and NF-κB.Conclusions MCP-1 may play an important role in the course of pancreatic fibrosis in chronic obstructive pancreatitis.

Isolation and cell culture of human bocavirus (HBoV) by human bronchial epithelial cell lines / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate pave a way for studying pathogenicty of HBoV.</p><p><b>METHODS</b>Isolation and cell culture of HBoV by human bronchial epithelial cell line, which was founded in our laboratory. The morphology of the virus were primarily studied with a transmission electron microscope. In addition, transcript mRNA was detected in human bronchial epithelial cells, which was passaged and infected within HBoV, using the reverse-transcription polymerase chain reaction (RT-PCR). The amplified products nucleotide sequence of HBoV were sequencing and sequence analysis.</p><p><b>RESULTS</b>Cytopathic effect (CPE) was observed after the aseptic residue of filtration of 2 case sputum specimens with HBoV, which was inoculated to the human bronchial epithelial cell line. The virus particles were observed in the cytoplasm, which were hexagonal or spherical in shape and 18-26 nm in diameter,bulk was 20 nm. cDNA amplicon obtained 295 bp fragment results of electrophoresis bands as same as NS1 region of the conserved matrix gene of publish sequence of HboV. PCR products nucleotide sequence of HboV were compared with corresponding HboV GeneBank sequences. The comparison/alignment and construction of phylogenetic trees also point to an affiliation of the parvovirus to the species HBoV.</p><p><b>CONCLUSION</b>Isolation and identification of HBoV could be done in the human bronchial epithelial cell, and we found some characterizing CPE in the human bronchial epithelial cell after HBoV infection. The above studies pave a way for studying pathogenicty of human bocavirus.</p>

Expression of recombinant VP2 gene in insect sf9 cells and screening of clinical specimens / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To clone and express VP, gene from HBoV, and the expressed VP, protein was as the antigen in order to detect serum from children in Wenling area with lower respiratory tract infections.</p><p><b>METHODS</b>The VP, gene was recombined with the genome of Baculovirus, which infected the insect cell. The fusion protein with HA tag was applied to confirm the specificity of expressed protein. Furthermore, the recombinant protein was observed using electron microscopy. The 176 serum from children in Wenling area with lower respiratory tract infections was screened using Western blot.</p><p><b>RESULTS</b>The expressed VP2 protein was more than 60% in total proteins from insect cell, and MWt about 60 x 10(3). The virus-like particle (VLP) was observed using electron microscopy, and size about 20 nm. The 176 serum from children in wenling area with lower respiratory tract infections was screened using Western blot. The HBoV positive rate was 2.28% (4/176).</p><p><b>CONCLUSION</b>The VP2 protein from human bocavirus was expressed in insect cell successfully. Through HA tag the VP2 protein was specific, and then the assay using SDS-PAGE with Western blot could detect and screen the antibody in serum from children with lower respiratory tract infections rapidly and accurately.</p>

Study on relationship between hepatitis B virus DNA load and genotype with large envelope protein / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the relation between hepatitis B virus DNA load and genotype with the level of large envelope protein.</p><p><b>METHODS</b>Serum HBV DNA was quantitively detected by using real time polymerase chain reaction (RT-PCR). The LHBs were detected by using enzyme linked immuno sorbent assay (ELISA) and HBV markers were detected by time differentiate immunofluorescence assay in 140 serum samples collected from chronic hepatitis B patients.The genotypes of HBV were identified by DNA sequencing; and analyze their relationship.</p><p><b>RESULTS</b>There was no significant difference between positive rate of LHBs and that of HBV DNA in HBeAg negative and positive group (P > 0.05); The HBV LHBs absorbency was markedly correlated with the HBV DNA load ( R2 = 0.9267). The difference of HBV LHBs absorbency between HBV genotype B and C was not significant.</p><p><b>CONCLUSIONS</b>The close correlation between HBV LHBs absorbence and HBV DNA load illustrated that he level of serum LHBs can be used to estimate the state of HBV replication; and there is no relationship between HBV LHBs absorbency and genotypes. So HBV LHBs may be used as a new serological marker to detect HBV replication.</p>