[Erdheim-Chester disease initially discovered at extraskeletal locations: a clinicopathological analysis of four cases].

Objective: To investigate the clinicopathological features of Erdheim-Chester disease (ECD) initially diagnosed at extraskeletal locations. Methods: Clinical and pathological data of four cases of ECD diagnosed initially in extraskeletal locations were collected at Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, from January 2013 to June 2023. BRAF V600E gene was detected by reverse transcription polymerase chain reaction (RT-PCR). Pertinent literatures were reviewed. Results: Four ECD patients included two males and two females ranging in ages from 2 years 11 months to 69 years. The lesions located in the lung (two cases), central nervous system (one case), and the testicle (one case) were collected in the study. One patient had occasional fever at night, one had nausea and vomiting, and two were asymptomatic. Radiologically, the two pulmonary ECD showed diffuse ground-glass nodules in both lungs, and the lesions in central nervous system and testicle both showed solid masses. Microscopically, there were infiltration of foamy histiocyte-like cells and multinucleated giant cells in a fibrotic background, accompanied by varying amounts of lymphocytes and plasma cells. The infiltration of tumor cells in pulmonary ECD was mainly seen in the subpleural area, interlobular septa, and perivascular and peribronchiolar areas. The fibrosis was more pronounced in the pleura and interlobular septa, and less pronounced in the alveolar septa. Immunohistochemical staining showed that all tumor cells expressed CD68, CD163 and Fô€ƒ¼a; one case showed S-100 expression; three cases were positive for BRAF V600E; all were negative for CD1α and Langerin. RT-PCR in all four cases showed BRAF V600E gene mutation. Conclusions: Extraskeletal ECD is often rare and occult, and could be easily misdiagnosed, requiring biopsy confirmation. The radiologic findings of pulmonary ECD is significantly different from other types of ECD, and the histopathological features of pronounced infiltration in the subpleura area, interlobular septa, perivascular and peribronchiolar areas can be helpful in the differential diagnosis from other pulmonary diseases. Detection of BRAF V600E gene mutation by RT-PCR and its expression by immunohistochemical staining are also helpful in the diagnosis.

[Comparison of QuantiFERON-TB Gold Plus and QuantiFERON-TB Gold In Tube in the diagnosis of pulmonary tuberculosis].

Objective: To compare the screening value of QuantiFERON-TB Gold Plus (QFT-Plus) and QuantiFERON®-TB Gold in tube (QFT-GIT) in the auxiliary diagnosis of pulmonary tuberculosis (TB). Methods: A screening test was performed. Patients who were hospitalized in Guangzhou Chest Hospital and underwent QFT-GIT testing from October to December 2020 were prospectively included as research subjects, QFT-Plus testing was added. And the basic information, clinical manifestations, laboratory test results, imaging examinations and other data of these patients were collected. A total of 207 patients were included and divided into tuberculosis group and non-tuberculosis group according to these data. There were 124 cases in the tuberculosis group (94 confirmed patients and 30 clinically diagnosed patients), including 90 males and 34 females, aged 18-93 years, with a median age of 57 (38, 67) years. The non-tuberculosis group included 83 patients (16 patients with non-tuberculous Mycobacteria and 67 patients with other lung diseases), including 49 males and 34 females, with a median age of 60 (51, 68) years. The confirmed patients were subdivided into three grades of low, medium and high Mycobacteriam tuberculosis (MTB) bacterial load, and three grades of mild, moderate and severe pulmonary tuberculosis. The results of QFT-Plus and QFT-GIT were compared, and the levels of IFN-γ in different antigen tubes were compared. Differences between different groups were compared using Mann-Whitney U test and Kruskal-Wallis H test. Results: The QFT-Plus showed a high degree of agreement with the QFT-GIT (κ=0.786, 95%CI: 0.740-0.832), while the main discordant result was QFT-GIT negative/QFT-Plus positive, accounting for 15/17. The sensitivity of QFT-GIT was 80.7%(95%CI: 0.706-0.880), the specificity was 76.3%(95%CI: 0.649-0.850), the positive predictive value was 79.8%(95%CI: 0.697-0.873), and the negative predictive value was 77.3%(95%CI: 0.659-0.859), repectively. QFT-Plus showed a sensitivity of 84.3%(95%CI: 0.743-0.910), a specificity of 78.8% (95%CI: 0.679-0.868), and a positive predictive value of 80.5%(95%CI: 0.703-0.879), the negative predictive value being 82.9%(95%CI: 0.721-0.902), slightly improved to that of the QFT-GIT. Also, this study found that there were significant differences in IFN-γ values between different MTB load or disease severity (P<0.05). Conclusions: There is a good consistency between the QFT-Plus test and the QFT-GIT test, both of which show good application value in the auxiliary diagnosis of pulmonary tuberculosis. Moreover, because of the addition of tuberculosis-specific CD8 cell antigen, the QFT-Plus test has higher sensitivity, lower uncertainty and more application value. This study also found that the bacterial load and disease severity of patients with pulmonary tuberculosis may have a certain correlation with the measured value of IFN-γ.

Effects of systemic therapies on pruritus in adults with atopic dermatitis: a systematic review and meta-analysis.

Pruritus is a hallmark of atopic dermatitis (AD), which affects disease severity and patient quality of life. In AD uncontrolled with first-line topical therapies or in moderate to severe AD, systemic therapies are used; however, there is a paucity of head-to-head trials comparing the effectiveness of these therapies. The aim of this study was to compare the effectiveness of systemic therapies in relieving pruritus in moderate to severe AD in adults, using a meta-analysis. The PubMed, EMBASE, Medline and CINAHL databases were searched from inception up to 31 May 2020 for randomized, placebo-controlled trials investigating the effectiveness of systemic therapies on pruritus with moderate to severe AD in patients aged ≥ 16 years. In total, 26 studies (n = 5190 participants) were identified. Compared with placebo, there was a large and statistically significant (P < 0.001 for all) reduction in pruritus [standard mean difference (SMD); 95% CI] with dupilumab every 2 weeks (-0.88; -1.13 to -0.63), dupilumab every 2 weeks plus topical corticosteroids (-0.77; -0.91 to -0.62), dupilumab once weekly (-0.99; -1.29 to -0.68), dupilumab once weekly plus topical corticosteroids (-0.70; -0.81 to -0.59). There was also a large and statistically significant reduction with ciclosporin (-1.30; -2.34 to -0.26; P = 0.01) and a large, although not statistically significant reduction with azathioprine (-0.85; -2.07 to 0.35). There was a small reduction with both mepolizumab (-0.27; -0.89 to 0.35) and interferon-γ (-0.31; -0.75 to 0.12). Of the investigational drugs, nemolizumab 2.0 mg/kg was the most effective (-8.13; -9.31 to -6.94). The majority of systemic therapies were superior to placebo in reducing pruritus. In particular, the dupilumab studies consistently showed large improvements in pruritus, while nemolizumab showed the strongest antipruritic effects. However, future head-to-head trials are required for conclusive evidence.

Epidemiology and Factors Affecting Functional Outcome of Distal Radial Fracture in an Urban Tertiary Medical Centre in Malaysia.

INTRODUCTION: Distal radial fracture is a commonly encountered fracture. This study aims to study the epidemiology of distal radial fracture and factors affecting the patients' functional outcome one to two years after the injury. MATERIALS AND METHODS: This is a retrospective cohort study. The records of patients, fulfilling the radiographical diagnosis of distal radial fracture, and aged 18 and above, who presented to our Emergency Department from 1st January 2018 to 31st December 2018 were retrieved. According to AO classification, we grouped our patients into A (extra-articular), B (partial articular) and C (complete articular). Patients with congenital abnormalities were excluded. Epidemiological data and relevant medical history were obtained and tabulated. A Malaysian language translation of Disability of the Arm, Shoulder and Hand (DASH) questionnaire was used to assess the functional outcome. RESULTS: Out of 168 patients' data retrieved, only 110 patients' data were found complete for purposes of this study. The mean DASH score was 13.7 ± 7.87 approximately one to two years post-injury regardless of treatment method. Increasing age was associated with higher DASH score with r=0.407(p<0.001). Several variables had significantly better functional outcome: male gender (p=0.01), Type A fracture configuration (p=0.007) and non-operational treatment (p=0.03). There was no significant difference between treatment modalities in Type A fracture (p=0.094), but Type B (p=0.043) and Type C (p=0.007) had better outcome without surgery. There was no significant difference between different ethnic groups, open or closed fracture and mechanism of injury. CONCLUSION: Better functional outcome after sustaining distal radial fracture was associated with young age, male gender, type A fracture and treated non-operatively. Interestingly, more complex fracture pattern had better functionality were observed without surgery.

[An advanced imaging method for measuring and assessing meibomian glands based on deep learning].

Objective: To evaluate the application value of a deep-learning-based imaging method for rapid measurement and evaluation of meibomian glands. Methods: Diagnostic evaluation study. From January 2017 to December 2018, 2 304 meibomian gland images of 576 dry eye patients who were treated at the Eye Center of Wuhan University People's Hospital with an average age of (40.03±11.46) years were collected to build a meibomian gland image database. These images were labeled by 2 clinicians, and a deep learning algorithm was used to build a model and detect the accuracy of the model in identifying and labeling the meibomian glands and calculating the rate of meibomian gland loss. Mean average precision (mAP) and validation loss were used to assess the accuracy of the model in identifying feature areas. Sixty-four meibomian gland images apart from the database were randomly selected and evaluated by 7 clinicians independently. The results were analyzed with paired t-test. Results: This model marked the meibomian conjunctiva (mAP>0.976, validation loss<0.35) and the meibomian gland (mAP>0.922, validation loss<1.0), respectively, thereby achieving high accuracy to calculate the area and ratio of meibomian gland loss. The proportion of meibomian glands marked by the model was 53.24%±11.09%, and the artificial marking was 52.13%±13.38%. There was no statistically significant difference (t=1.935, P>0.05). In addition, the model took only 0.499 second to evaluate each image, while the average time for clinicians was more than 10 seconds. Conclusion: The deep-learning-based imaging model can improve the accuracy of the examination and save time and be used for clinical auxiliary diagnosis and screening of diseases related to meibomian gland dysfunction.(Chin J Ophthalmol, 2020, 56: 774-779).

[Current situation of screening, prevention and treatment of bleeding esophageal varices in cirrhotic portal hypertension in Tibet region: a multicenter study].

Objective: To investigate and analyze the current situation, screening, clinical characteristics, prevention and treatment of bleeding esophageal varices in cirrhotic patients with portal hypertension in Tibet region. Methods: Clinical data of cirrhotic patients with portal hypertension through March 2017 to February 2020 from Tibet region were collected and analyzed retrospectively. Results: 511 cases with liver cirrhosis were included in the study, of which 185 cases (36.20%) had compensated cirrhosis and 326 cases (63.80%) had decompensated cirrhosis. Further analysis of the etiological data of liver cirrhosis showed that 306 cases (59.88%) were of chronic hepatitis B, 113 cases (22.11%) of alcoholic liver disease, and 68 cases (13.31%) of chronic hepatitis B combined with alcoholic liver disease. Among patients with compensated liver cirrhosis, 48 cases (25.95%) underwent endoscopic examination of which 33 diagnosed as high-risk variceal bleeding. However, none of these 33 cases had received non-selective ß-blocker therapy, and only four patients had received endoscopic variceal banding therapy. Among patients with decompensated liver cirrhosis, 83 cases (25.46%) had a history of upper gastrointestinal bleeding, 297 cases (91.10%) had ascites, 23 cases (7.05%) had hepatic encephalopathy, and 3 cases (0.92%) had hepatorenal syndrome. Among the patients with a history of upper gastrointestinal bleeding, 42 cases (50.60%) had received secondary preventive treatment for bleeding esophageal varices, including 39 cases of endoscopic treatment, 1 case of endoscopic combined drug treatment, 3 cases of interventional treatment, and 2 cases of surgical treatment. Conclusion: Chronic hepatitis B and alcoholic liver diseases are the main causes of liver cirrhosis in Tibet region. Moreover, this region lacks screening, prevention and treatment for bleeding esophageal varices in cirrhotic patients with portal hypertension. Therefore, it is necessary to increase the screening of high-risk groups to prevent and improve the first-time bleeding, and promote multidisciplinary team to prevent and treat re-bleeding.

[The ratio of tuberculosis-specific antigen to phytohemagglutinin in T-SPOT assay in the diagnosis of active tuberculosis].

Objective: The aim of this study was to determine the performance of the ratio of tuberculosis-specific antigen (TBAg) to phytohemagglutinin (PHA) (TBAg/PHA ratio) in T-SPOT assay in the diagnosis of active tuberculosis (ATB). Methods: Between January 2014 and January 2017, 378 Mycobacterium tuberculosis (MTB) culture positive patients (268 cases of pulmonary tuberculosis, 110 extra-pulmonary tuberculosis) and 824 healthy individuals were recruited from Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. T-SPOT assay was performed and TBAg/PHA ratio was calculated in all the participants. To validate the study, another group of 223 MTB culture positive TB patients with positive T-SPOT results were recruited from Guangzhou Chest Hospital between January 2017 and December 2017. This was a retrospective case-control study and differences between groups were analyzed using the Mann-Whitney U-test. Results: Of the 378 culture positive ATB patients, 344 patients had positive T-SPOT results. Of the 824 healthy individuals, 204 individuals had positive T-SPOT results. Using healthy individuals as the control group, the sensitivity and specificity of T-SPOT assay in the diagnosis of ATB were 91.0% (344/378) and 75.2% (620/824). Directly using T-SPOT results had a limited accuracy in distinguishing ATB from latent tuberculosis infection (LTBI). The area under the receiver operating characteristic (ROC) curve was between 0.7 and 0.8. However, a further calculation of the TBAg/PHA ratio showed a better performance than TBAg in distinguishing these two conditions, and the area under the ROC curve was 0.881 (95% CI: 0.853-0.909). If using the threshold value of 0.234, the sensitivity and specificity of the TBAg/PHA ratio in distinguishing ATB from LTBI were 69.5% (239/344) and 94.12% (192/204). The validation data showed that the performance of the TBAg/PHA ratio in distinguishing ATB from LTBI was also satisfactory, and the area under the ROC curve was 0.901 (95% CI: 0.872-0.931). Furthermore, the TBAg/PHA ratio had an important role in the diagnosis of extra-pulmonary tuberculosis. If using the threshold value of 0.234, the sensitivity and specificity of the TBAg/PHA ratio in the diagnosis of extra-pulmonary tuberculosis were 79.2% (76/96) and 94.1% (192/204). The area under the ROC curve was 0.932 (95% CI: 0.897-0.967). Conclusions: The TBAg/PHA ratio in T-SPOT assay was better than directly using T-SPOT results in distinguishing ATB from LTBI. This ratio also showed a potential use in the diagnosis of extra-pulmonary tuberculosis.

Detection rate of colonic polyp among patients who had undergone colonoscopy at gastroenterology unit of Serdang Hospital, Malaysia.

OBJECTIVE: The aim of this study was to evaluate the demography, and to determine the detection rate of polyps, and detection rate of adenoma at a Malaysian tertiary hospital. METHODS: This is a retrospective study of all the patients who had undergone colonoscopy at Gastroenterology endoscopy unit, Serdang Hospital from 1st January 2010 to 31st December 2016. Patients who had a history of colorectal cancer, polyp or inflammatory bowel disease were excluded. Data collected which included patients' demography, indication for colonoscopy, colonoscopy finding, and histopathology results. Data was analysed with SPSS version 16. RESULTS: Among the 559 patients who had fulfilled the inclusion criteria (68 males, 44 females), 112 patients were found to have at least one polyp giving the polyp detection rate (PDR) of 20% and 168 polypectomies were performed. The PDR among male patients was higher than that of females (22.5% vs 17.1%, p<0.05). The detection rate of polyp was nearly equal in Malays, Chinese, Indians, and Others. The polyps were more common in those of age 40 years old and above (p<0.05), with the mean age of 63.0±1.5 years. The commonest morphology of polyp in our patients was sessile (58%) and majority was medium size (5-9mm). Otherwise, the polyps were commonly found in the distal colon those that in proximal colon (55.3% vs 38.7%, p<0.05). The adenoma detection rate (ADR) was 19.1% (107/559). CONCLUSION: The detection rate of colonic polyp from colonoscopy is 20% in our centre.

MiR-218 suppresses metastasis and invasion of endometrial cancer via negatively regulating ADD2.

OBJECTIVE: Endometrial cancer (EC) is one of the three most common gynecological cancers. Due to the lack of effective treatment for EC patients in an advanced stage, the mortality rate of EC is increasing rapidly. Hence, it is essential to seek for novel molecular therapeutic targets and biomarkers for EC. The aim of this study was to explore the role of miR-218 in the occurrence and development of EC and to investigate the possible underlying mechanism. PATIENTS AND METHODS: The expression level of miR-218 in EC tissues and cell lines was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Wound healing assay and Matrigel assay were performed to determine the migration and invasion abilities of EC cells. Meanwhile, the potential targets of miR-218 were predicted by bioinformatics analysis and confirmed by Luciferase reporter gene assay. In addition, the protein expression level of Adducin 2 (ADD2) was assessed by Western blotting analysis. RESULTS: QRT-PCR results revealed that miR-218 was significantly downregulated in EC tissues and cell lines. Wound healing assay and Matrigel assay demonstrated that miR-218 suppressed the migration and invasion abilities of EC cells. Online prediction databases predicted that ADD2 was a direct target of miR-218, which was verified by Luciferase reporter gene assay. Rescue experiments further validated that miR-218 could serve as a carcinoma suppressor by negatively regulating ADD2 expression in EC. CONCLUSIONS: In the present study, we elucidated that miR-218 served as a tumor suppressor in EC by negatively regulating ADD2. This might bring a novel insight into new molecular therapeutic targets and biomarkers for EC.

Expressions of VEGF and miR-21 in tumor tissues of cervical cancer patients with HPV infection and their relationships with prognosis.

OBJECTIVE: To investigate the expressions of vascular endothelial growth factor (VEGF) and micro-ribonucleic acid-21 (miR-21) in cervical cancer patients with human papillomavirus (HPV) infection and determine the potential relationships with prognosis. PATIENTS AND METHODS: Expressions of VEGF in cervical cancer tissues and cancer-adjacent tissues were detected by immunohistochemistry, and the expressions of miR-21 and VEGF in both tissues were quantitatively analyzed using reverse transcription polymerase chain reaction (RT-PCR). Patients with cervical cancer were followed up after operation, and the survival rates of patients with different expression levels of miR-21 and VEGF were compared. RESULTS: VEGF was expressed in both cervical cancer tissues and cancer-adjacent tissues. The positive expression rate of VEGF in cervical cancer tissues (75.69%) was significantly higher than that in cancer-adjacent tissues (10.45%). RT-PCR results showed that the expression levels of miR-21 and VEGF in cervical cancer tissues were significantly higher than those in cancer-adjacent tissues (p<0.05). Correlation analyses revealed that miR-21 expression was significantly positively correlated with VEGF expression in cervical cancer tissues (r2=0.4174, p<0.0001). Prognostic analyses showed that the 5-year survival rate of patients was relatively high when miR-21 and VEGF were lowly expressed. CONCLUSIONS: VEGF and miR-21 are highly expressed in tumor tissues of cervical cancer patients with HPV infection. VEGF expression is significantly positively correlated with miR-21 expression, and the high levels of VEGF and miR-21 predict unfavorable prognosis of cervical cancer. Data provide a theoretical support for clinical treatment of cervical cancer patients with HPV infection.

[Effects of zinc ions on biological functions of human umbilical vein endothelial cells].

Objective: To evaluate the effect of zinc ions on human umbilical vein endothelial cells biological functions. Methods: The primary human umbilical vein endothelial cells were cultured with the ECM medium, and cells were divided into 8 groups: the control group(routine culture,n=3), 20 µmol/L zinc group(20 µmol/L zinc chloride solution was added into the cell medium, n=3), 40 µmol/L zinc group(40 µmol/L zinc chloride solution was added into the cell medium, n=3),80 µmol/L zinc group(80 µmol/L zinc chloride solution was added into the cell medium, n=3), 100 µmol/L zinc group(100 µmol/L zinc chloride solution was added into the cell medium, n=3), 200 µmol/L zinc group(200 µmol/L zinc chloride solution was added into the cell medium, n=3),300 µmol/L zinc group(300 µmol/L zinc chloride solution was added into the cell medium, n=3), 500 µmol/L zinc group(500 µmol/L zinc chloride solution was added into the cell medium, n=3). The cell proliferation curve was derived from real time cell analysis (RTCA). The viability value was obtained via CCK-8 reagent, and the migration distance was tested by scratch-wound assay while the adhesion function was detected by RTCA. Results: (1)After 18 hours, RTCA showed that the proliferation cell indexes were 4.5±0.6, 3.7±0.4, 3.6±0.3, 2.5±0.4, and 2.5±0.4 in the 20, 40, 80, 100, and 200 µmol/L zinc groups, as compared with 3.5±0.3 in the control group (all P<0.05). Proliferation cell indexes were 0 in both of the 300 µmol/L and 500 µmol/L zinc groups. (2)After 96 hours, the viability were 1.21±0.05, 1.10±0.03, 0.99±0.05, 0.62±0.02, 0.45±0.04, 0.11±0.01, and 0.12±0.06, respectively in the 20, 40, 80, 100, 200, 300, and 500 µmol/L zinc groups, as compared with 0.75±0.05 in the control group (all P<0.05). (3)After 12 hours, the migration distances were (0.56±0.11),(0.96±0.07),(0.49±0.02), and (0.29±0.01)mm in the 20, 40, 80, and 100 µmol/L zinc groups, as compared with (0.24±0.04)mm in the control group (all P<0.05). (4)After 18 hours, the adhesion cell index were 0.40±0.05, 0.31±0.01, 0.38±0.05, and 0.40±0.03 in the 20, 40, 80, and 100 µmol/L zinc groups, as compared with 0.24±0.04 in the control group (all P>0.05). Conclusions: Zinc ions at lower concentration (≤80 µmol/L) can promote proliferation, viability and migration of human umbilical vein endothelial cells, but the adhesion function was not significantly affected by zinc ions. Zinc ions at higher concentration (≥200 µmol/L) can inhibit the cellular function of the human umbilical vein endothelial cells.

A physiologically relevant 3D collagen-based scaffold-neuroblastoma cell system exhibits chemosensitivity similar to orthotopic xenograft models.

3D scaffold-based in vitro cell culturing is a recent technological advancement in cancer research bridging the gap between conventional 2D culture and in vivo tumours. The main challenge in treating neuroblastoma, a paediatric cancer of the sympathetic nervous system, is to combat tumour metastasis and resistance to multiple chemotherapeutic drugs. The aim of this study was to establish a physiologically relevant 3D neuroblastoma tissue-engineered system and explore its therapeutic relevance. Two neuroblastoma cell lines, chemotherapeutic sensitive Kelly and chemotherapeutic resistant KellyCis83 were cultured in a 3D in vitro model on two collagen-based scaffolds containing either glycosaminoglycan (Coll-GAG) or nanohydroxyapatite (Coll-nHA) and compared to 2D cell culture and an orthotopic murine model. Both neuroblastoma cell lines actively infiltrated the scaffolds and proliferated displaying >100-fold increased resistance to cisplatin treatment when compared to 2D cultures, exhibiting chemosensitivity similar to orthotopic xenograft in vivo models. This model demonstrated its applicability to validate miRNA-based gene delivery. The efficacy of liposomes bearing miRNA mimics uptake and gene knockdown was similar in both 2D and 3D in vitro culturing models highlighting the proof-of-principle for the applicability of 3D collagen-based scaffolds cell system for validation of miRNA function. Collectively, this data shows the successful development and characterisation of a physiologically relevant, scaffold-based 3D tissue-engineered neuroblastoma cell model, strongly supporting its value in the evaluation of chemotherapeutics, targeted therapies and investigation of neuroblastoma pathogenesis. While neuroblastoma is the specific disease being focused upon, the platform may have multi-functionality beyond this tumour type. STATEMENT OF SIGNIFICANCE: Traditional 2D cell cultures do not completely capture the 3D architecture of cells and extracellular matrix contributing to a gap in our understanding of mammalian biology at the tissue level and may explain some of the discrepancies between in vitro and in vivo results. Here, we demonstrated the successful development and characterisation of a physiologically relevant, scaffold-based 3D tissue-engineered neuroblastoma cell model, strongly supporting its value in the evaluation of chemotherapeutics, targeted therapies and investigation of neuroblastoma pathogenesis. The ability to test drugs in this reproducible and controllable tissue-engineered model system will help reduce the attrition rate of the drug development process and lead to more effective and tailored therapies. Importantly, such 3D cell models help to reduce and replace animals for pre-clinical research addressing the principles of the 3Rs.

Metallic powder-bed based 3D printing of cellular scaffolds for orthopaedic implants: A state-of-the-art review on manufacturing, topological design, mechanical properties and biocompatibility.

Metallic cellular scaffold is one of the best choices for orthopaedic implants as a replacement of human body parts, which could improve life quality and increase longevity for the people needed. Unlike conventional methods of making cellular scaffolds, three-dimensional (3D) printing or additive manufacturing opens up new possibilities to fabricate those customisable intricate designs with highly interconnected pores. In the past decade, metallic powder-bed based 3D printing methods emerged and the techniques are becoming increasingly mature recently, where selective laser melting (SLM) and selective electron beam melting (SEBM) are the two representatives. Due to the advantages of good dimensional accuracy, high build resolution, clean build environment, saving materials, high customisability, etc., SLM and SEBM show huge potential in direct customisable manufacturing of metallic cellular scaffolds for orthopaedic implants. Ti-6Al-4V to date is still considered to be the optimal materials for producing orthopaedic implants due to its best combination of biocompatibility, corrosion resistance and mechanical properties. This paper presents a state-of-the-art overview mainly on manufacturing, topological design, mechanical properties and biocompatibility of cellular Ti-6Al-4V scaffolds via SLM and SEBM methods. Current manufacturing limitations, topological shortcomings, uncertainty of biocompatible test were sufficiently discussed herein. Future perspectives and recommendations were given at the end.

Bioavailability of butachlor and myclobutanil residues in soil to earthworms.

To establish chemical extraction procedures for predicting bioavailability of butachlor and myclobutanil in soil, several solvent systems, including methanol, methanol-water (9:1), methanol-water (1:1), acetone-water (5:3), petroleum ether and water, were assessed for their feasibility in determining extractability of the target compounds from soil samples. Experimental data showed that the extractability of butachlor and myclobutanil by the solvents was well linearly correlated with their bioavailability to Eisenia foetida and Allolobophora caliginosa, indicating that these extraction procedures may be efficient for predicting bioavailability of the two pesticides. The concentrations of the pesticides accumulated in E. foetida and A. caliginosa varied with species, suggesting that the availability of the soil-sequestered pesticide is a species-dependent process.

Correlation between serum level of neuron-specific enolase and long-term functional outcome after acute cerebral infarction: prospective study.

OBJECTIVE: To determine the value of measuring serum levels of neuron-specific enolase in predicting extent of disease and short- and long-term functional outcome after acute cerebral infarction. DESIGN: Prospective study. SETTING: Neurology departments at two university teaching hospitals, Shanghai. PATIENTS: Thirty-eight patients who presented for acute cerebral infarction between October 1998 and October 2000 were divided into two groups: those whose infarction extended to the cerebral cortex in the carotid artery region (cortical group) and those with an infarction in the subcortical carotid artery region (subcortical group). MAIN OUTCOME MEASURES: Using a solid-phase enzyme immunoassay, we measured serum levels of neuron-specific enolase on admission and on days 2, 3, and 15. Infarct volume was measured by computed tomography on day 5. The Activities of Daily Living scale was used to assess the clinical outcome at 1-, 3-, and 6-month follow-up after onset. RESULTS: Mean (standard deviation) serum neuron-specific enolase levels were significantly higher among patients with acute cerebral infarction than among controls (18.48 [16.61] ng/mL versus 9.00 [2.70] ng/mL; P<0.001). The neuron-specific enolase level was also higher in the cortical group than in the subcortical group (33.54 [29.71] ng/mL versus 15.97 [5.91] ng/mL; P<0.01). Levels peaked after 2.11 (0.86) days and correlated positively with the infarct volume (r=0.81; P<0.01) and negatively with clinical outcome at 1 month (r= -0.37; P<0.05), 3 months (r= -0.45; P<0.01), and 6 months (r= -0.65; P<0.001), as assessed on the Activities of Daily Living scale. CONCLUSION: Serum neuron-specific enolase levels after cerebral infarction may be a useful marker to predict infarct volume and short- or long-term functional outcome.

Protein folding from a highly disordered denatured state: the folding pathway of chymotrypsin inhibitor 2 at atomic resolution.

Previous experimental and theoretical studies have produced high-resolution descriptions of the native and folding transition states of chymotrypsin inhibitor 2 (CI2). In similar fashion, here we use a combination of NMR experiments and molecular dynamics simulations to examine the conformations populated by CI2 in the denatured state. The denatured state is highly unfolded, but there is some residual native helical structure along with hydrophobic clustering in the center of the chain. The lack of persistent nonnative structure in the denatured state reduces barriers that must be overcome, leading to fast folding through a nucleation-condensation mechanism. With the characterization of the denatured state, we have now completed our description of the folding/unfolding pathway of CI2 at atomic resolution.

The hepatitis C virus core protein interacts with NS5A and activates its caspase-mediated proteolytic cleavage.

Viral proteins interact with one another during viral replication, assembly, and maturation. Systematic interaction assays of the hepatitis C virus (HCV) proteins using the yeast two-hybrid method have uncovered a novel interaction between core and NS5A. This interaction was confirmed by in vitro binding assays, and coimmunoprecipitation in mammalian cells. Core and NS5A are also colocalized in COS-7 cells. Interestingly, NS5A is cleaved to give specific-size fragments, when core is coexpressed in mammalian cells. Overexpression of core produced many dying and rounded cells and effects such as DNA laddering and the truncation of poly(ADP-ribose) polymerase 1 (PARP1), both indicators of apoptosis. These observations led us to investigate the link between the induction of apoptosis by core and the cleavage of NS5A. The proteolysis of NS5A and these apoptotic events can be inhibited by caspase inhibitor, Z-VAD, indicating that core induces apoptosis and the cleavage of NS5A by caspases. In cells infected by the HCV, core may provide the intrinsic apoptotic signal, which produces truncated forms of NS5A. The biological function of core-NS5A interaction and the downstream effect of NS5A cleavage are discussed.

Inactivation of the human papillomavirus-16 E6 oncoprotein by organic disulfides.

We are investigating compounds that could be useful in the treatment of neoplastic lesions of the cervix by acting on the oncoprotein E6 of human papillomavirus-16. The E6 protein contains two potential zinc-binding domains that are required for most of its functions. We have published tests that measure (i) the release of zinc ions after chemical alteration of the cysteine groups of these zinc-binding domains (TSQ assay), (ii) the interaction of E6 with the cellular proteins E6AP and E6BP (BIACORE assay), and (iii) the viability of tumor cell lines that require the continuous expression of HPV oncoproteins (WST1 assay). Based on these tests, we identified 4.4'-dithiodimorpholine as a potential lead compound. In this study we examined whether the dithiobisamine moiety of 4,4'-dithiodimorpholine may be an important molecular prerequisite for further drug development in this system. We have evaluated 59 new substances including organic disulfides and those containing the dithiobisamine moiety, as well as structural analogues. The compounds with significant reactivity in all three assays were observed only for dithiobisamine derivatives with saturated cyclic amines and aryl substituted piperazines. The identity of these substances suggests that the N-S-S-N moiety is necessary but not sufficient for reactivity in our assays, and that dithiobisamine based substances are useful as lead compounds that target the cysteine groups of HPV-16 E6 zinc fingers.