The level of Tim-3+CD8+ T cells can serve as a potential marker for evaluating the severity of acute graft-versus-host disease after haplo-PBSCT

Early and accurate diagnosis of acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation is crucial for the prognosis of patients. This study identified a potential biomarker for the severity of aGVHD after human leukocyte antigen (HLA)-haploidentical peripheral blood hematopoietic stem cell transplantation (haplo-PBSCT). We included 20 healthy subjects and 57 patients who underwent haplo-PBSCT. Of these patients, 22 developed aGVHD after haplo-PBSCT. The results showed that patients with aGVHD had significantly increased levels of Tim-3+/Perforin+/Granzyme B+CD8+ T cells, but significantly decreased Galectin-9. The differences in Galectin-9 and Tim-3+/Granzyme B+CD8+ T cells between grade I-II aGVHD and III-IV aGVHD were also significant. In vitro, the apoptosis of CD8+ T cells from aGVHD patients was significantly increased after Tim-3/Galectin-9 pathway activation, which decreased Granzyme B secretion. As revealed by univariate analysis, the level of Tim-3+CD8+ T cells was a risk factor for severe aGVHD. ROC analysis demonstrated that high levels of Tim-3+CD8+ T cells had a significant diagnostic value for severe aGVHD, with an area under the curve of 0.854 and cut-off value of 14.155%. In conclusion, the binding of Tim-3 with exogenous Galectin-9 can promote apoptosis of CD8+ T cells and affect the secretion of Granzyme B. Tim-3+CD8+ T cells have the potential to serve as immunological markers for assessing the severity of aGVHD after haplo-PBSCT and identifying patients at a higher risk for severe aGVHD.

Prevalence of Clonorchis sinensis infections in freshwater fish in mainland China: A meta-analysis / 中国血吸虫病防治杂志

Objective To understand the real prevalence of Clonorchis sinensis infections in the freshwater fish in mainland China, so as to provide insights into clonorchiasis control and detection of freshwater fish. Methods All literatures reporting the prevalence of C. sinensis infections in the freshwater fish, the second intermediate host of the parasite, were jointly retrieved in Chinese and English electronic databases from January 1, 2010 to December 31, 2020, including Wanfang Data, CNKI, PubMed, Web of Science, Embase and Cochrane Library. All studies were screened based on inclusion and exclusion criteria, and the quality of all enrolled literatures was evaluated. The pooled prevalence of C. sinensis infections in freshwater fish and its 95% confidence interval (CI) were estimated using the software Stata version 15.0, and subgroup analyses were performed to investigate the region-, season- and sample source-specific pooled prevalence of C. sinensis infections in freshwater fish. In addition, the sensitivity and publication bias of all included studies were analyzed. Results A total of 40 eligible literatures were included in this study, including 37 Chinese literatures and 3 English literatures, and there were 10 high-quality literatures, 27 moderate-quality literatures and 3 low-quality literatures. A total of 53 species containing 37 959 freshwater fish were reported in these 40 studies, and 73.58% (39/53) of freshwater fish species were identified with C. sinensis infections. Meta-analysis showed 23.5% [95% CI: (0.19, 0.28)] pooled prevalence of C. sinensis infections in freshwater fish in mainland China, and subgroup analyses higher prevalence of C. sinensis infections in freshwater fish in northeastern China [35.7%, 95% CI: (0.22, 0.50)] than in central [25.9%, 95% CI: (0.04, 0.48)] and southern China [20.6%, 95% CI: (0.09, 0.32)], higher prevalence of C. sinensis infections in freshwater fish sampled in spring [44.1%, 95% CI: (0.35, 0.53)] than in autumn [6.7%, 95% CI: (0.05, 0.08)] and summer [3.3%, 95% CI: (−0.01, 0.07)], and higher prevalence of C. sinensis infections in freshwater fish sampled from natural water [25.2%, 95% CI: (0.17, 0.33)] than from retail trades [22.2%, 95% CI: (0.17, 0.28)] and breeding chain [12.3%, 95% CI: (0.03, 0.22)]. However, all included studies had a publication bias with a low sensitivity. Conclusions The prevalence of C. sinensis infections is high in freshwater fish in mainland China, and there are still challenges for clonorchiasis control. Reinforcement of health education, diagnostics development and food safety supervision is recommended in future clonorchiasis control programs.

COL6A3 promotes cellular malignancy of osteosarcoma by activating the PI3K/AKT pathway

SUMMARY OBJECTIVE In this study, we aimed to investigate the role of COL6A3 on cell motility and the PI3K/AKT signaling pathway in osteosarcoma. METHODS The relative expression of COL6A3 was achieved from a GEO dataset in osteosarcoma tissue. siRNA technology was applied to decrease the COL6A3 expression in cells, and cell counting kit-8 (CCK-8) assay and colony formation analysis were used to examine the cell proliferation potential. Knockdown COL6A3 made the proliferation and colony formation abilities worse than the COL6A3 without interference. Likewise, in contrast to the si-con group, cell invasion and migration were inhibited in the si-COL6A3 group. Moreover, the western blot results suggested that the PI3K/AKT signaling pathway was manipulated by measuring the protein expression of the PI3K/AKT pathway-related markers, due to the COL6A3 inhibition. CONCLUSION COL6A3 plays a crucial role in modulating various aspects of the progression of osteosarcoma, which would provide a potentially effective treatment for osteosarcoma.

RESUMO OBJETIVO Neste estudo, investigamos a função do COL6A3 na mobilidade celular e na via PI3K/AKT em osteossarcomas. METODOLOGIA A expressão relativa do COL6A3 foi obtida a partir de dados GEO em tecidos de osteossarcoma. O RNA de interferência (siRNA) foi utilizado para reduzir a expressão do COL6A3 nas células, e o teste de contagem de células kit-8 (CCK-8) e a análise de formação de colônias foram realizados para examinar o potencial de proliferação celular. Além disso, o Transwell comprovou os efeitos do si-COL6A3 na invasão celular e migração em células de osteossarcoma. Para medir os níveis de expressão das proteínas e mRNAs, utilizamos transcriptase reversa quantitativa (qRT-PCR) e western blot. RESULTADOS O COL6A3 foi regulado nos tecidos e células do osteossarcoma quando comparado com o controle normal. A redução de COL6A3 reduziu a proliferação e a capacidades de formação de colônias em relação ao COL6A3 sem interferência. Do Mesmo modo, ao contrário do observado no grupo si-con, a invasão e migração celular foram inibidas no grupo si-COL6A3. Além disso, o resultado do western blot sugere que a via PI3K/AKT foi manipulada, medindo a expressão proteica dos marcadores relacionados à PI3K/AKT, devido à inibição do COL6A3. CONCLUSÃO O COL6A3 desempenha um papel crucial na modulação de vários aspectos da progressão do osteossarcoma, o que pode representar um possível tratamento eficaz para a doença.

Factors influencing suicidal ideation and suicide attempt of middle school students in minority areas of Guizhou Province / 中国学校卫生

Objective@#To analyze the prevalence and influencing factors of suicidal behaviors among middle school students in minority areas of Guizhou Province, and to provide the reference for targetedly preventing suicidal behavior among middle school students.@*Methods@#Questionnaire survey including adolescent health information, Depression Self-rating Scale for Children(DSR-SC), Childhood Trauma Questionnaire-Short Form (CTQ-SF) and the Pittsburgh Sleep Quality Index survey were conducted among 6 139 middle school students in minority areas of Guizhou Province, Logistic regression was used to analyze the influencing factors of suicidal behavior of middle school students.@*Results@#A total of 1 241(20.2%) had suicidal ideation in the past 12 months, and 250 (4.1%) of middle school students had committed suicide in the past 12 months. Suicidal ideation among middle school students was positively associated with: female, non-residence, being only child, smoking, having poor relationship with teachers, poor relationship with classmates, school violence victimization, physical abused, sexual abuse, emotionally neglected, having depression, having sleeping disorders(OR=1.67, 1.64, 0.84, 0.72, 0.67, 0.58, 2.06, 2.45, 2.12, 1.68, 2.82, 1.62). While being women(OR=1.52), non-residence(OR=2.25), smoking(OR=0.52), poor relationship with teachers(OR=0.49), poor relationship with classmates(OR=0.47), school violence victimization (OR=1.79) and physically abused(OR=2.17), sexual abused(OR=1.97), physically neglected(OR=1.94), depressive symptoms(OR=3.14), sleep disorders (OR=1.66) were associated positively with the risk of suicide attempt.@*Conclusion@#Suicide-related behaviors are more common in minority areas of Guizhou Province. Teachers and parents should attach great importance to it, and their suicidal risk should be early recognized to reduce suicide-related behaviors.

An investigation and analysis of health-related quality of life in patients with Kashin-Beck disease in Shaanxi Province / 中华地方病学杂志

Objective To investigate health-related quality of life (HRQOL) and influencing factors in adult patients with Kashin-Beck disease (KBD) in Shaanxi Province for improving health-related quality of life of KBD patients.Methods By the two-stage cluster random sampling method,data of 901 cases of KBD patients with the disease severity of grade 1 and higher were collected from higher KBD incidence areas of Baoji,Xianyang and Tongchuan in Shaanxi Province in February 2016.The Chinese version of SF-36 (including 8 dimensions:physical functioning,role-physical,bodily pain,general health,vitality,social functioning,role-emotional,mental health) was used to measure the HRQOL.Multiple linear regression was used to analyze the influencing factors of HRQOL.Results In 901 cases of KBD patients,male accounted for 55.49％ (500/901),female accounted for 44.51 (401/901).The median age of patients was 58 (22-83) years old.According to the severity of disease criteria,patients with grade Ⅰ were 37.18％ (335/901),patients with grade Ⅱ were 53.50％ (482/901) and patients with grade Ⅲ] were 9.32％ (84/901).The scores of each dimension of SF-36 scale in patients of male patients had higher scores in GH and MH than women [GH:35 (25,50) vs 30 (20,45),MH:56 (48,72) vs 56 (44,68),.9 ＜ 0.05].There were significant differences in PF,RP,BP,GH,VT and SF scores among different age groups (P ＜ 0.05).Similarly,there were significant differences in PF,RP,GH,VT,SF,RE and MH scores among patients with different educational levels (P ＜ 0.05).Married group was only higher in the PF and SF scores than other marital status groups [PF:55 (40,75) vs 50 (35,65),SF:62 (50,75) vs 50 (38,75),P ＜ 0.05].As expected,there were significant differences in the scores of PF,BP,GH and SF between the severity of diseases.There were also significant differences in the scores of PF,BP,GH,VT,SF and MH in KBD patients from different regions (P ＜ 0.05).According to multiple linear regression analysis results,male was the conservation factor of GH and MH scores compared with female;age was a risk factor for PF,RP,BP,GH,VT and SF scores;compared with the illiterate,primary school education was a protective factor for PF and RE scores,junior high school and above were PF,RP,VT,SF,RE and MH scores protective factors;compared with degree Ⅰ KBD,degree Ⅱ and degree Ⅲ were risk factors for PF,BP,GH and SF scores;compared with Baoji area,Xianyang area was a protective factor of SF score,however,Tongchuan area was a protective factor for PF,GH,VT,SF and MH scores.Conclusions KBD has severely damaged the HRQOL of patients.Priority should be given to age,educational attainment and severity of the disease for developing measures to improve KBD patients' HRQOL in different regions.

Subxiphoid video-assisted thoracoscopic thymectomy versus traditional video-assisted thoracic surgery thymectomy for myasthenia gravis: A case control study / 中国胸心血管外科临床杂志

@#Objective To investigate the clinical outcomes of subxiphoid video-assisted thoracoscopic thymectomy for myasthenia gravis. Methods The clinical data of the 85 patients undergoing video-assisted thoracoscopic thymectomy for myasthenia gravis in Department of Cardiothoracic Surgery, Huashan Hospital affiliated to Fudan University between January 2014 and July 2016 were studied. Subxiphoid approach video-assisted thoracoscopic thymectomy (SXVT) and through traditional unilateral approach video-assisted thymectomy (TVAT) were compared. The clinical outcomes of SXVT and TVAT were compared. Results There was no surgical death and no statistical difference between the two groups in drainage time, postoperative volume of drainage, postoperative hospital stay and bleeding volume during operation (P>0.05). However, the acute chest pain after surgery, as well as the postoperative chest pain, and operative time were less in the the SXVT group than that in the TVAT group (P<0.05). Conclusion SXVT for myasthenia gravis is safe and executable. It can alleviate intercostal neuralgia and abnormal chest wall feeling. And it should be considered in the treatment of myasthenia gravis.

Application of Leucocyte Adherence Inhibition Test in Determining Immunological Activity of Ribonucleic Acid / 中国药师

Objective:To establish a method for determining the immunological activity of ribonucleic acid. Methods: Leucocyte adherence inhibition test ( LAI) was applied, and the important parameters of LAI including the mouse strain, drug concentration, treatment time, content of buffer solution and cell density were researched. The immunological activity of RNAⅠ, Ⅱand Ⅲ was re-spectively determined by the method. Results:Stable and reliable parameters were obtained: the sample concentration was 10 mg· ml-1 , the treatment time was 2 hours, Ca2+ and Mg2+ were necessary for the buffer solution, and the cell density was about 4 × 107 cell·ml-1 . The strain of mouse showed no effect on the results. As a result, the determination method for immunological activity was established. Using the method, the immunological activity of RNA Ⅰ,Ⅱand Ⅲ was determined 3 times, and the results met the re-quirements with RSD below 20%. Conclusion:The method is suitable for determining the immunological activity of RNA.

Rare cavernous hemangioma of adrenal gland: case report / Hemangioma cavernoso raro da glândula adrenal: relato de caso

CONTEXT: Cavernous hemangiomas of the adrenal gland are rare benign neoplastic tumors. The clinical presentation of adrenal hemangiomas is usually vague, and they are often discovered incidentally through imaging examination s performed for other reasons. CASE REPORT: We report the case of a non-functional adrenal hemangioma found incidentally in a 37-year-old man with a one-year history of headache and hypertension. A right adrenal mass was detected by means of magnetic resonance imaging. Physical examination and all laboratory values were unremarkable. The patient underwent laparoscopic right adrenal gland resection. Histopathological evaluation confirmed adrenal cavernous hemangioma. CONCLUSIONS: Most occurrences of cavernous hemangiomas of the adrenal gland are non-functional and often discovered incidentally. Although rare, these unusual benign adrenal masses should form part of the differential diagnosis of adrenal neoplasms. The proper treatment for adrenal cavernous hemangioma is surgical removal. .

CONTEXTO: Hemangiomas cavernosos da glândula adrenal são tumores neoplásicos raros, benignos. A apresentação clínica dos hemangiomas adrenais é geralmente vaga e muitas vezes eles são descobertos acidentalmente por exame imagiológico realizado por outras razões. RELATO DE CASO: Nós relatamos um caso de hemangioma adrenal não funcional encontrado por acaso em um homem de 37 anos de idade com histórico de um ano de dor de cabeça e hipertensão. Foi detectada massa adrenal direita por meio de ressonância magnética. Exame físico e todos os valores de laboratoriais estavam normais. O paciente foi submetido a cirurgia laparoscópica para ressecção da glândula adrenal direita. Avaliação histopatológica confirmou hemangioma adrenal cavernoso. CONCLUSÃO: A maioria dos hemangiomas cavernosos de glândula adrenal são não funcionais e muitas vezes descobertos por acaso. Embora raras, essas massas adrenais benignas devem ser parte de um diagnóstico diferencial de neoplasias suprarrenais. O tratamento adequado para o hemangioma adrenal cavernoso é a remoção cirúrgica. .

A family cluster of nitrite poisoning, Suzhou City, Jiangsu Province, China, 2013

Background：In April 2013, a hospital in Suzhou City notified authorities of a patient with nitrite poisoning with two other family members who had similar toxic symptoms five days prior. We investigated the event to identify the cause, source and possible route of contamination.Methods：A case was defined as any person living in the Yang Shan Hua Yuan community who had been diagnosed with cyanoderma and food poisoning symptoms from 15 to 25 April 2013. Active case finding was conducted by interviewing community residents and reviewing medical records from local clinics; information was then retrospectively collected on the patient’s food history, cooking procedures and food sources.Results：We identified three nitrite poisoning cases, one male and two females, from the same family. The time between dinner and onset of illness was less than an a hour. A retrospective survey showed that a substance presumed to be sugar mixed with asparagus on 17 April and with stir-fried asparagus on 21 April was the suspected contaminant. The presumed sugar came from a clean-up of a neighbouring rental house. Nitrite was detected in a vomitus sample, the sugar substance and two leftover food samples.Conclusion：This family cluster of nitrite poisoning resulted from the mistaken use of nitrite as sugar to cook dishes. We recommend that sodium nitrite be dyed a bright colour to prevent such a mistake and that health departments strengthen food hygiene education to alert people about the danger of eating unidentified food from an unknown source.

In vitro and in vivo studies of pirarubicin-loaded SWNT for the treatment of bladder cancer

Intravesical chemotherapy is an important part of the treatment for superficial bladder cancer. However, the response to it is limited and its side effects are extensive. Functional single-walled carbon nanotubes (SWNT) have shown promise for tumor-targeted accumulation and low toxicity. In the present study, we performed in vivo and in vitro investigations to determine whether SWNT-based drug delivery could induce high tumor depression in rat bladder cancer and could decrease the side effects of pirarubicin (tetrahydropyranyl-adriamycin, THP). We modified SWNT with phospholipid-branched polyethylene glycol and constructed an SWNT-THP conjugate via a cleavable ester bond. The cytotoxicity of SWNT-THP against the human bladder cancer cell line BIU-87 was evaluated in vitro. Rat bladder cancer in situ models constructed by N-methyl-N-nitrosourea intravesical installation (1 g/L, 2 mg/rat once every 2 weeks for 8 weeks) were used for in vivo evaluation of the cytotoxicity of SWNT and SWNT-THP. Specific side effects in the THP group including urinary frequency (N = 12), macroscopic hematuria (N = 1), and vomiting (N = 7) were identified; however, no side effects were observed with SWNT-THP treatment. Flow cytometry was used to assess the cytotoxicity in vitro and in vivo. Results showed that SWNT alone did not yield significant tumor depression compared to saline (1.74 ± 0.56 and 1.23 ± 0.42%) in vitro. SWNT-THP exhibited higher tumor depression than THP-saline in vitro (74.35 ± 2.56 and 51.24 ± 1.45%) and in vivo (52.46 ± 2.41 and 96.85 ± 0.85%). The present findings indicate that SWNT delivery of THP for the treatment of bladder cancer leads to minimal side effects without loss of therapeutic efficacy. Therefore, this nanotechnology may play a crucial role in the improvement of intravesical treatment of bladder cancer.

Breast Cancer Associated Fibroblasts Promote MCF-7 Invasion in vitro by Secretion of HGF / 华中科技大学学报（医学）（英德文版）

This study was aimed to explore the influence of breast cancer associated fibroblasts (CAFs) in migration and invasion of breast cancer cell line MCF-7,and investigate whether hepatocyte growth factor (HGF) is involved in this process.Primary breast CAFs and their corresponding normal breast fibroblasts (NFs) were obtained by collagenase digestion.On the basis of the co-culture,the migration and invasion capacity of MCF-7 cells was compared between CAFs and NFs by Transwell.The difference in the HGF expression between them was detected by ELISA.The secretion of HGF was knocked down by using RNA interference technology in CAFs.Then the changes of migration and invasion capacity of MCF-7 cells were investigated by Transwell.Eventually,we isolated high-purity CAFs and NFs,and the CAFs had a stronger ability in promoting MCF-7 migration and invasion than the NFs.ELISA results demonstrated that CAFs secreted higher HGF,and the capacity of MCF-7 migration and invasion was declined after knocking down the secretion of HGF in CAFs by RNA interference.It is suggested that CAFs can promote MCF-7 migration and invasion through HGF in vitro.

Silencing MTA1 by RNAi Reverses Adhesion, Migration and Invasiveness of Cervical Cancer Cells (SiHa) via Altered Expression of p53, and E-cadherin/β-catenin Complex / 华中科技大学学报（医学）（英德文版）

It has been reported that metastasis-associated gene 1 (Mtal) is overexpressed in many malignant tumors with high metastatic potential.In addition,some studies indicated that MTA1 participated in invasion,metastasis,and survival of cancer cells by regulating cell migration,adhesion and proliferation.But the role of MTA 1 is unclear in vitro in the development of cervical cancer cells.This study investigated whether and how MTA1 mediated cell proliferation,migration,invasion and adhesion in cervical cancer.MTA1 expression level was detected by Western blot in two cervical cancer cell lines of different invasion potentials.The effects of MTA1 expression on SiHa cell apoptosis,cycle,proliferation,migration,invasion and adhesion were tested by flow cytometry,MTT,wound-healing assay,Transwell assay and adhesion assay,respectively.The expression levels of p53,E-cadherin,and β-catenin activity were evaluated in untreated and treated cells.The results showed that MTA1 protein expression was significantly higher in SiHa than in HeLa,which was correlated well with the potential of migration and invasion in both cell lines.Furthermore,the cell invasion,migration and adhesion capabilities were decreased after inhibition of MTA1 expression mediated by Mtal-siRNA transfection in SiHa.However,no significant differences were found in cell apoptosis,cycle,and proliferation.In addition,E-cadherin and p53 protein levels were significantly up-regulated,while β-catenin was significantly down-regulated in SiHa transfected with the siRNA.These results demonstrated that MTA1 played an important role in the migration and invasion of cervical cancer cells.It was speculated that the decreased migration and invasion capability by inhibiting the MTA1 expression in the SiHa cell line may be mediated through the altered expression of p53,and E-cadherin/β-catenin complex.MTA1 could serve as a potential therapeutic target in cervical cancer.

miR-125b Confers Resistance of Ovarian Cancer Cells to Cisplatin by Targeting Pro-apoptotic Bcl-2 Antagonist Killer 1 / 华中科技大学学报（医学）（英德文版）

Chemotherapy is the preferred therapeutic approach for advanced ovarian cancer,but a suceessful long-term treatment is prevented by the development of drug resistance.Recent works have underlined the involvement of non-coding RNAs,microRNAs (miRNAs) in cancer development,with several conjectures regarding their possible involvement in the evolution of drug resistance.This study is to investigate the promoting effects and mechanism of miR-125b involved in the development of chemoresistance in ovarian cancer.The different expression of miR-125b in cisplatin-sensitive ovarian cancer cell line (OV2008) and its resistant variant (C13*) was identified by real-time PCR.An in vitro cytotoxicity assay and apoptosis assay using CCK-8 assay and flow cytometry,were carried out to detect the effect of miR-125b and Bak1 on cisplatin resistance of cells.Real-time PCR,Western blotting and luciferase reporter assay were used to detect whether Bak1 is a target of miR-125b.As compared with OV2008 cells,the expression levels of miR-125b in C13* cells were increased.It was found that the up-regulation of microRNA-125b caused a marked inhibition of cisplatin-induced cytotoxicity and apoptosis and a subsequent increase in the resistance to cisplatin in OV2008 and C13* cells.Moreover,Bakl was a direct target of miR-125b,and down-regulation of Bak1 suppressed cisplatin-induced apoptosis and led to an increased resistance to cisplatin.Our study indicates that miR-125b has a significantly promoting effect on chemoresistance of C13* cells and up-regulation of miR-125b expression contributes to cisplatin resistance through suppression of Bakl expression.This finding has important implications in the development of targeted therapeutics for overcoming cisplatin resistance in ovarian cancer.

Adenocarcinoma of esophago-gastric junction / 国际外科学杂志

The incidence of gastroesophageal junction cancers is rapidly increasing, the Year' s upwards rate being more than 5％ ～ 10％. Siewert in 1987 proposed a definition for adenocarcinoma of the esophagogastric junction (AEG), as a adenocarcinoma within 5 cm of the gastroesophageal junction (GEJ), and divided AEG in to three types. The majority of patients with AEG in Asian countries have type Ⅱ and Ⅲ cancers, and no obvious differences have been reported in 5-year survival rates between patients with different types of AEG. In contrast, in western countries, the distribution of each type of AEG is nearly equal,patients with AEG type Ⅰ cancer have the best prognosis, whereas the overall survival of patients with type Ⅲ cancer is the worst. Now people presume that the AEG is a special type of carcinoma which is not same as carcinoma of esophagus and stomach. The surgical routes and operative methods and the extent of lymph node dissection of AEG are still not completely clear, and AEG lacks also a independent TNM system for itself. We will summarize the actuality of diagnosis and therapy of AEG.

Preparation and assay of C-glucosyltransferase from roots of Pueraria lobata.

C-glucosyltransferase (EC 2.4.1.X) is one of the key enzymes for the biosynthesis of puerarin. This paper describes the methodology in purification and assay of the enzyme for the first time in Puerarin lobata (Willd.) Ohwi. C-glucosyltransferase from roots of P. lobata was extracted and partially purified by (NH4)2SO4 saturation. The effects of pH, temperature, and substrate concentration on the activity of the enzyme were investigated. The properties of the puerarin produced by C-glucosyltransferase were studied by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The peak activity of C-glucosyltransferase was detected in fraction of by 80% saturation of (NH4)2SO4 and the optimal conditions for enzymatic reaction were 35.5 μmol l-1 of isoliquiritigenin and 560 μmol l-1 of UDP-G at pH 8.1, 28oC for 1 h. Mn2+ at 1 mmol l-1 and Al3+ at 1 mmol l-1 increased the enzyme activity, while Mg2+ inhibited its activity. The enzyme activity in Nicotiana tabacum and P. lobata were detected under the above assay conditions. Higher activity was found in roots than in leaves and stems of P. lobata, while no enzyme activity was detected in leaves of N. tabacum. It was the first time that activity of C-glucosyltransferase, which transforms isoliquiritigenin to puerarin, was detected in P. lobata.

Effect of Spindle Checkpoint on Akt2-mediated Paclitaxel-resistance in A2780 Ovarian Cancer Cells / 华中科技大学学报（医学）（英德文版）

Recent evidence has suggested that Akt2 plays an important role in the protection of cells from paclitaxel(PTX)-induced apoptosis and control of the cell cycle.In addition,some scholars suggested that the PTX sensitivity depends on a functional spindle assembly checkpoint.In the present study,we investigated the role of the Akt2/Bub1 cross-talking in apoptosis and cell cycle after exposure of the A2780 ovarian cancer cells to paclitaxel(PTX).Recombinant expression plasmid WT-Akt2 was transfected into A2780 cells by lipofectamine2000,and then the expression level of Akt2 gene was detected by using RT-PCR and Western blotting.Cell apoptosis and cell cycle were detected by flow cytometry and Hoechst 33342 staining after treatment with PTX.Moreover,we compared the expression level of Bubl in different groups by Western blotting.Our study showed that up-regulation of Akt2 contributed to A2780 ovarian cancer cells overriding PTX-induced G2/M arrest,and inhibited Bub1 expression.Our findings might shed light on the molecular mechanism of PTX-induced resistance in ovarian cancer and help develop novel anti-neoplastic strategies.

Suppression of Allogeneic T Cells Proliferation by CD3/CD46-Induced T-regulatory 1 Cells / 华中科技大学学报（医学）（英德文版）

CD46 is not only identified as a complement regulatory protein which protects host cells from complement attack,but also a new co-stimulatory molecule for human T cells.CD3/CD46 co-stimulation can induce a T-regulatory 1 cell(Tr1)-specific cytokine phenotype in human CD4+T cells.However,the role of CD46 as a co-stimulatory molecule in the modulation of the acquired immunity,such as transplant immunology,remains unclear.In this study,CD4+T cells were isolated from human CD46-transgenic C57BL/6 mice by magnetic-activated cell sorting,and further induced by anti-CD3,anti-CD28 and anti-CD46 antibodies respectively,and anti-CD3/anti-CD28 antibodies,anti-CD3/anti-CD46 antibodies,or the monoclonal antibody panel against CD3/CD28/CD46.The levels of interleukin-2(IL-2),γ-interferon(γ-IFN),interleukin-10(IL-10)and transforming growth factor-β(TGF-β)were detected in the supematants of different groups.Suppression of allogeneic T cell proliferation were assessed by using mixed lymphocyte reaction(MLR)assay,in which monoclonal antibodies against CD46 were added to the culture.The results showed that CD3/CD28,CD3/CD46 and CD3/CD28/CD46 co-stimulation could significantly induce stronger proliferation of T cells than CD3 stimulation(P＜0.05),and CD3/CD28/CD46 co-stimulation significantly increased the proliferation of T cells when compared with CD3/CD28 or CD3/CD46 co-stimulation(P＜0.05 for each).IL-2 and γ-IFN levels were much higher in CD3/CD28 co-stimulation group than in CD3,CD28,CD46 and CD3/CD46 groups(P＜0.05 for each).IL-10 and TGF-β levels were dramatically increased in CD3/CD46 co-stimulation group as compared with those in the CD3,CD28,CD46 and CD3/CD28 groups(P＜0.05 for each).CD3/CD46 co-stimulation significantly inhibited the T cell proliferation and allogenic immune responses through the secretion of IL-10 and TGF-β in MLR (P＜0.05).These results suggested that CD3/CD46 can induce Tr1 cells to modulate allogenic immune responses,and it may become a novel target for the development of new therapeutic approach for T-cell-mediated diseases.CD46 plays an important role in regulating the T cell-mediated immune responses by bridging innate and acquired immunity.

Treatment of anterior dislocation of sternoclavicular joint with Kirschner wire minimally invasive fixation / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the safety and efficacy of close reduction and percutaneous needle fixation for the treatment of anterior dislocation of sternoclavicular joint.</p><p><b>METHODS</b>A retrospective analysis was performed with 6 cases of anterior sternoclavicular dislocation by close reduction and percutaneous needle fixation with Kirschner wire treated from January 2001 to February 2009, including 5 males and 1 female aged from 19 to 45 with an average of 28.8 years old. Among the 6 cases, 4 were on right lateral and 2 were on left lateral. The time from injured to treatment was from 6 hours to 12 days (averaged 4.5 days). The clinical effects were evaluated according to Rockwood scoring and the complications were observed.</p><p><b>RESULTS</b>All 6 patients were followed-up for 3 to 13 months (averaged 6 months). According to Rockwood scoring,the preoperative score was (7.00 +/- 0.89) points, postoperative score was (13.17 +/- 1.72) points; the results showed excellent in 5 cases and good in 1 case. No local infection, postoperative pain,recurrent dislocation,broken needle, and other complications were observed in this study.</p><p><b>CONCLUSION</b>The treatment of anterior sternoclavicular joint dislocation with Kirschner wire minimally invasive fixation is an easy, reliable fixation with less complications.</p>

Expression of insulin-like growth factor-1 and alkaline phosphatase in the reconstruction of cleft palate with distraction osteogenesis in rhesus / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the osteogenesis mechanism by analysis of the expression of insulin-like growth factor-I (IGF-1) and alkaline phosphatase (ALP) in the reconstruction of cleft palate (CP) with distraction osteogenesis (DO) in rhesus.</p><p><b>METHODS</b>The CP animal models were established surgically. 21 rhesus in experimental group underwent DO to close the soft and bony defect, followed by consolidations. Every 3 animals were killed and the specimen were taken out after consolidation of 1, 2, 4, 6, 8, 12, 24 weeks. The mRNA of IGF-1 and ALP were detected with Real-time RT-PCR technique. The expression of IGF-1 and ALP was quantitatively analyzed by ELISA. The results were compared with those in control and sham groups (each of 2 animals), respectively.</p><p><b>RESULTS</b>Since consolidation, the mRNA of IGF-1 and ALP increased significantly at one week and reached the peak at two weeks, but decrease to control level after 12 weeks of consolidation. The expression of IGF-1 also increased to peak level after two weeks of consolidation. The expression of ALT increased significantly since consolidation and reach the peak value after six weeks. They all decreased to nearly control level after 8-12 weeks.</p><p><b>CONCLUSIONS</b>The palate cleft can be successfully closed with new formed bone after DO. The mechanism of bone consolidation is intramembranous bone formation.</p>

Expression of decoy receptor 3 in liver tissue microarrays.

Background. Decoy receptor 3 (DcR3), a new member of the tumour necrosis factor receptor (TNFR) superfamily, is amplified and overexpressed in various cancers. We investigated the expression of DcR3 protein in liver tissue microarrays and assessed its importance in patients with hepatocellular carcinoma (HCC). Methods. In this retrospective study, tissue from 120 patients with HCC, 48 with tissue at least 2 cm away from the tumour (juxta-tumour tissue), 62 with cirrhosis and 23 with normal livers were studied as tissue microarrays. Immunohistochemistry was used to detect the expression of DcR3. Statistical analyses were done to assess the association between DcR3 expression and the clinicopathological features of HCC. Results. The positivity rate of DcR3 in HCC tissue was significantly higher than that in juxta-tumour tissue, cirrhosis and normal liver (p=0.017, p<0.0001, p<0.0001, respectively). The positive rate of DcR3 in juxta-tumour and cirrhotic tissue both increased significantly when compared with normal liver tissue (p<0.0001, p=0.005, respectively). The positivity rate of DcR3 in HCC in clinical TNM stages I and II was significantly lower than that in stages III and IV (p<0.0001). The positivity rate of DcR3 in patients without metastasis within 20 months decreased significantly compared with those with metastasis (p<0.0001). DcR3 expression in patients with alphafoetoprotein levels >400 g/L, portal vein tumour emboli, capsular infiltration and multicentric tumour was significantly higher than in groups without these features (p=0.021, p<0.0001, p<0.0001, p=0.002, respectively). Conclusion. The overexpression of DcR3 might play an important role in the pathogenesis, progression and metastases of HCC. The DcR3 gene might serve as an important molecular biological indicator in diagnosing and predicting the biological behaviour of patients with HCC.