Tertiary Lymphoid Structure Raises Survival and Immunotherapy in HPV- HNSCC.

Immune checkpoint blockade (ICB) targeting PD-1/PD-L1 has been used for the treatment of head and neck squamous cell carcinoma (HNSCC). However, the overall response rate to ICB therapy for HNSCC remains less than 20%. It has recently been reported that the appearance of tertiary lymphoid structures (TLSs) in tumor tissue is correlated with better prognosis and response to ICB treatment. Here, we demonstrated an immune classification for the tumor microenvironment (TME) of HNSCC by analyzing The Cancer Genome Atlas (TCGA)-HNSCC data set and found that immunotype D with TLS enrichment had a better prognosis and response to ICB treatment. Furthermore, we observed that TLSs were present in a part of tumor samples of human papillomavirus (HPV) infection negative HNSCC (HPV- HNSCC) and were associated with the densities of dendritic cell (DC)-LAMP+ DCs, CD4+ T cells, CD8+ T cells, and progenitor T cells in TME. We established an HPV- HNSCC mouse model with TLS-enriched TME by overexpressing LIGHT in a mouse HNSCC cell line. We found that the induction of TLS formation enhanced the response to PD-1 blockade treatment in the HPV- HNSCC mouse model, accompanied by increases in DCs and progenitor exhausted CD8+ T cells in the TME. Elimination of CD20+ B cells attenuated the therapeutic effect of PD-1 pathway blockade in TLS+ HPV- HNSCC mouse models. These results indicate that TLSs contribute to the favorable prognosis and antitumor immunity of HPV- HNSCC. Inducing TLS formation in HPV- HNSCC tumors is a potential therapeutic method for improving the ICB response rate in patients with HPV- HNSCC.

[Clinical characteristics of 14 cases of neonatal tracheotomy in neonatal intensive care unit].

Objective: To analyze the clinical characteristics of neonatal tracheotomy in neonatal intensive care unit (NICU). Methods: This single-center retrospective study included 14 neonates admitted to NICU of Beijing Children's Hospital, Capital Medical University from January 2016 to August 2021, and were<28 days of age on admission, who met the criteria of tracheotomy and finally completed the procedure. The clinical characteristics including age, weight, duration of ventilation, etiology of tracheotomy, length of hospital stay and prognosis were summarized and analyzed. Wilcoxon signed-rank test was used to compare the weight gain velocity and the duration of ventilation before and after tracheotomy. Paired t-test was used to compare the hospitalization length before and after tracheotomy. Spearman correlation was used to analyze the correlation between the clinical characteristics and outcomes. Results: For the 14 neonates, the gestational age was (38±4) weeks and birth weight was (2 824±949) g. Nine of them were male. The age on transportation was 16 (6, 25) d. A total of 10 neonates were on invasive ventilation on admission, the other 4 were on nasal continuous positive airway pressure support. Bilateral vocal cord paralysis (7 cases) was the commonest cause of tracheotomy. The age on operation was 33 (22, 44) d. There were statistically significant differences in duration of ventilation and weight gain velocity before and after operation (19.00 (10.50, 34.00) vs. 0.86 (0.06, 3.25) d, 1.66 (-0.16, 5.54) vs. 4.69 (2.30, 9.32) g/(kg·d), Z=3.01 and -1.98, both P<0.05). The total hospital stay in NICU was (37±12) d. One neonate died during hospitalization. The existence of pneumonia on admission was positively correlated to NICU stay length (r=0.57, P=0.027), the pre-operational weight gain velocity was negatively correlated to the post-operational NICU stay length (r=-0.73, P=0.020). There were 4 neonates de-cannulated during 7-38 months after the tracheotomy, and 5 neonates still wearing the tracheal cannulation during 15-66 months after the tracheotomy. Two neonates died and 2 neonates lost follow-up after discharge. All neonates could not vocalize normally before de-cannulation, and the language development obviously lagged behind the normal age group after de-cannulation. Conclusions: Bilateral vocal cord paralysis is the commonest cause of neonatal tracheotomy. The benefit of tracheotomy for NICU neonates with surgical indications is obvious, especially in facilitating extubation and improving weight gain.

Steinernema populi n. sp. (Panagrolaimomorpha, Steinernematidae), a new entomopathogenic nematode species from China.

Steinernema populi n. sp. was recovered by baiting from beneath poplar trees in China. Morphological and molecular features provided evidence for placing the new species into the Kushidai clade. The new species is characterized by the following morphological features: third-stage infective juveniles (IJ) with a body length of 1095 (973-1172) µm, a distance from the anterior end to excretory pore of 77 (70-86) µm and a tail length of 64 (55-72) µm. The Body length/Tail length (c) ratio and Anterior end to Excretory pore/ Tail length × 100 (E%) of S. populi n. sp. are substantially greater than those of all other 'Feltiae-Kushidai-Monticolum' group members. The first-generation males can be recognized by a spicule length of 66 (57-77) µm and a gubernaculum length of 46 (38-60) µm. The new species is further characterized by sequences of the internal transcribed spacer and partial 28S regions of the ribosomal DNA. Phylogenetic analyses show that Steinernema akhursti and Steinernema kushidai are the closest relatives to S. populi n. sp.

[Introduction of landmarking approach and its application in dynamic prediction].

Conventional prediction model, as a static prediction model, can be only used to predict the probability of the occurrence of an event during the observation period using the information available at baseline survey. However, based on current clinical demands, dynamic prediction, which obtains prediction probabilities for both baseline survey and later time points given the history of the events and covariates up to that time, is gaining a growing attention. As a dynamic prediction model, the landmarking approach is simple, easy to use, computationally efficient and has a comparable performance of joint modeling, which makes it to be widely used in recent researches. Because of its limited application in China, this paper makes a brief introduction of its ideas and basic application to further promote its applications in clinical dynamic prediction.

[Clinical characteristics and outcomes of 111 neonates with upper airway obstruction admitted via transportation].

Objectives: To analyze the clinical characteristics and outcomes of neonates with upper airway obstruction (UAO) who were admitted via transportation, hence to provide more evidence-based information for the clinical management of UAO. Methods: This was a single center retrospective study. Patients were hospitalized in Beijing Children's Hospital from January 1, 2016 to May 31, 2021 with age <28 days or postmenstrual age (PMA) ≤44 weeks, and UAO as the first diagnosis. The general information of patients, obstructed sites in the upper airway, treatment, complications and prognosis were analyzed. The outcomes of surgical UAO vs. non-surgical UAO were analyzed by 2 by 2 χ2 test. Results: A total of 111 cases were analyzed (2.3% of the total NICU hospitalized 4 826 infants in the same period), in which 62 (55.9%) were boys and 101 (91.0%) were term infants, and their gestational age was (38.7±2.0) weeks, birth weight (3 207±585) g, PMA on admission (40.8±2.5) weeks and weight on admission was (3 221±478) g. There were 92 cases (82.9%) with symptoms of UAO presenting on postnatal day 1, and 35 cases (31.5%) had extra-uterine growth retardation on admission. The diagnosis of UAO and the obstructive site was confirmed in 25 cases (22.5%) before transportation. There were 24 cases (21.6%), 71 cases (64.0%), and 16 cases (14.4%) who had UAO due to nasal, throat, and neck problems, respectively. The top 5 diagnosis of UAO were vocal cord paralysis (28 cases), bilateral choanal atresia (20 cases), laryngomalacia (15 cases), pharynx and larynx cysts (7 cases), and subglottic hemangioma (6 cases). The diagnosis and treatment of all the patients followed a multidisciplinary approach consisted of neonatal intensive care unit, ear-nose-throat department and medical image departments. A total of 102 cases (91.9%) underwent both bronchofiberscope and fiber nasopharyngoscope investigation. Seventy cases (63.1%) required ventilation. Among the 58 cases (52.3%) who required surgical intervention, 16 had tracheotomy. For cases with vs. without surgical intervention, the rate of cure and (or) improvement were 94.8% (55/58) vs. 54.7% (29/53), and the rate of being discharged against medical arrangement were 1.7% (1/58) vs. 45.3% (24/53) (χ²=24.21 and 30.11, both P<0.01). Conclusions: Neonatal UAO may locate at various sites of the upper airway. The overall prognosis of neonatal UAO is favorable. A multidisciplinary approach is necessary for efficient evaluation and appropriate surgical intervention.

[A study on the online medical consulting websites based on the personal computer side].

Online medical consulting, acting as the primary type of Internet medical market, has been developing dramatically in the past ten years and begins to take shape. This study collected available information to describe service content status and service provider for online medical consulting websites. The current online medical consulting sites are mainly comprehensive medical consultation websites. The most common consulting provision from is combining graphics and text, which might not meet users' primary demand. The registered physicians are mostly the ones with junior position and work in the eastern and south-central parts of China. Activities of the registered physicians vary across the departments, but with extremely low initiatives.

Virtual reality for acute and chronic pain management in adult patients: a narrative review.

Virtual reality is a computer-generated environment that immerses the user in an interactive artificial world. This ability to distract from reality has been utilised for the purposes of providing pain relief from noxious stimuli. As technology rapidly matures, there is potential for anaesthetists and pain physicians to incorporate virtual reality devices as non-pharmacological therapy in a multimodal pain management strategy. This systematic narrative review evaluates clinical studies that used virtual reality in adult patients for management of acute and chronic pain. A literature search found 690 citations, out of which 18 studies satisfied the inclusion criteria. Studies were assessed for quality using the Jadad and Nottingham-Ottawa Scales. Agreement on scores between independent assessors was 0.87 (95%CI 0.73-0.94). Studies investigated virtual reality use: intra-operatively; for labour analgesia; for wound dressing changes; and in multiple chronic pain conditions. Twelve studies showed reduced pain scores in acute or chronic pain with virtual reality therapy, five studies showed no superiority to control treatment arms and in one study, the virtual reality exposure group had a worsening of acute pain scores. Studies were heterogeneous in: methods; patient population; and type of virtual reality used. These limitations suggest the evidence-base in adult patients is currently immature and more rigorous studies are required to validate the use of virtual reality as a non-pharmacological adjunct in multimodal pain management.

[Reevaluation of equivocal HER2 status detected by immunohistochemistry according to the 2019 guidelines for HER2 detection].

Objective To understand the effects and clinical significance of the 2019 guidelines for the human epidermal growth factor receptor 2 (HER2) detection. Methods: According to the 2014 guidelines, 548 cases of invasive breast cancer with equivocal HER2 (2+) detected by immunohistochemistry (IHC) in Taizhou Enze Medical Center, Zhejiang Province, China from 2013 to 2019 were selected. The results of IHC and HER2/CEPl7 double-probe were reevaluated and divided into groups according to the 2019 guidelines for the comparative analysis. Results: Among the 548 IHC HER2 (2+) invasive breast cancers, the number of positive, equivocal and negative cases for HER2 were 96 (17.52%), 81 (14.78%) and 371 (67.70%), respectively, according to the 2014 guidelines. However, according to the 2019 guidelines, 10 cases (1.82%) were reclassified as IHC 1+, 2 cases in the group 2 were reclassified as negative, and all the originally equivocal cases in group 4 were reclassified as negative. Finally, the total number of positive and negative cases for HER2 were 94 (17.15%) and 454 (82.85%), respectively. Conclusions: After applying the 2019 guidelines, the number of IHC 2+ cases decreases, and the positive rate for HER2 also decreases slightly due to the reevaluation change in groups 2 and 4, leading to reclassification of the cases that were deemed equivocal according to the 2014 guidelines. In general, the new 2019 guidelines are more reasonable and easier to use.

[Application of Python web crawler technology in infodemiology].

Python web crawler technology, which automatically and massively getting information from the Internet by mimicking net users' browsing behavior, is a basic supporting technique to extract and integrate multi-source heterogeneous data in the field of Infodemiology. There are two types of Python web crawler: simple and massive-scale, both collect information simultaneously from the database establishment. Advantages of this technique are characterized as: being simple syntax, in high flexibility and low cost in learning and maintenance. Contents of the current application scenarios include surveillance, implementation and evaluation of health intervention programs on public health issues, as well as on smart doctor seeking. For the last two years, the Chinese government started to encourage the integration and utilization of multi-source heterogeneous data including internet information. Hence, the number of application scenarios for Python web crawler technology are bound to increase in the foreseeable future. Corresponding matched talent cultivations and technical innovations are suggested to add to the current education and research systems on public health issues.

[Analysis of robotic natural orifice specimen extraction surgery on 162 cases with rectal neoplasms].

Objective: To explore the safety and feasibility of da Vinci robot surgical systems in natural orifice specimen extraction surgery (NOSES) for rectal neoplasms. Methods: A descriptive cohort study was used. Inclusion criteria: (1) age ≥18 years old; (2) diagnosis of rectal cancer by biopsy via colonoscopy or benign neoplasm locating in rectum that could not be resected locally through the anus; (3) R0 resection can be achieved by preoperative evaluation; (4) the CDmax (maximum circumferential diameter) was ≤5 cm or specimens could still be extracted from the anus despite a CDmax exceeding 5 cm but was along the longitudinal axis of the rectum. Exclusion criteria: (1) emergency operation due to gastrointestinal obstruction, perforation, or bleeding; (2) distal metastasis, induding lung, bone, or liver, that could not be resected simultaneously; (3) history of abdominal surgery or any other contraindications for robotic surgery. Clinicopathological data of 162 patients with rectal neoplasms who underwent robotic NOSES at the General Surgery Department of the Second Xiangya Hospital of Central South University from March 2016 to July 2019 were retrospectively collected. Of 162 patients, 94 were male and 68 were female; the average age was (57±13) years; the average BMI was (23.5±3.2) kg/m(2); the average distance from tumor to the anal verge was (8.2±2.9) cm. Five trocars were used to perform total mesorectal excision (TME), and the descending colon artery was preserved. Sterile endoscope sleeve for the specimen extraction was inserted into the pelvic cavity through the anus, and the resected specimen was pulled out through the sleeve. Outcomes of safety (operation time, intraoperative blood loss and postoperative morbidity of complication) and oncological outcomes (number of lymph nodes harvested, rate of lymph node metastasis and rate of positive resection margin) were collected. Results: All the 162 cases completed robotic NOSES successfully with no conversion to laparotomy. The average operation time was (188.7±79.8) minutes; the average blood loss was (47.1±33.2) ml; the average and the maximum CDmax of specimens were (3.4±1.5) cm and 12 cm respectively. A total of 154 patients underwent robotic TME. One underwent robotic TME plus resection of liver metastasis; one underwent robotic TME plus partial transverse colectomy; two patients underwent robotic TME plus ovariectomy; another two underwent robotic TME plus hysterectomy; one patient underwent robotic TME plus left partial nephrectomy due to renal angioleiomyoma; another one underwent robotic TME plus ureteral repair due to intraoperative injury of the left ureter. All the specimens were extracted through the anus. Protective ileostomy was performed in 6.8% (11/162) of the patients. The average number of lymph node harvested was 14.9±5.1. According to pathological reports, 156 neoplasms were adenocarcinoma. Tis stage was 1.3% (2/156), T1 stage was 9.0% (14/156), T2 stage was 26.3% (41/156), T3 stage was 35.9% (56/156), and T4 stage was 27.6% (43/156). Lymph node metastasis accounted for 34.6% (54/156), and simultaneous liver metastasis was observed in one case. Circumferential resection margins (CRMs) and upper and lower resection margins were negative in all the patients. The average postoperative feeding time and postoperative hospital stay were (4.2±4.1) days and (11.4±7.7) days, respectively. Postoperative morbidity of complication was 12.3% (20/162). The incidence of anastomotic leakage was 4.9% (8/162), of which only 4 cases (2.5%) received ileostomy. Within postoperative 90-day, no anal dysfunction or death were found. Conclusion: Robotic NOSES for rectal neoplasms is safe and feasible.

[The performance of visual photoscreening for Chinese preschool children aged 4 to 5 years].

Objective: To assess the accuracy of photoscreening for detecting refractive amblyopia risk factors (ARFs) in Chinese preschool children aged 4 to 5 years. Methods: A cross-sectional study. Comprehensive ocular examinations were conducted for preschool children in Nanjing, China from September to December, 2016. Photoscreening (Plusoptix A12C) was applied for refractive screening without cycloplegia. Voluntary children and children suspected of eyes abnormalities received cycloplegic retinoscopy (CR). Results of photoscreening and CR were compared using Wilcoxon signed rank test, and Bland-Altman plot were used to assess the agreement between the photoscreener and CR. According to the updated preschool vision screening guidelines from American Association for Pediatric Ophthalmology and Strabismus (AAPOS) in 2013, CR was adopted for identifying children with ARFs, which was considered as a golden standard. Based on the golden standard, the accuracy of 5 sets of referral criteria (including sensitivity standard, Matta/Silbert standard, AAPOS2013 standard, Alaska Blind Child Discovery standard, specificity standard) for photoscreener were tested. Receiver operating characteristics curves were constructed applied to evaluate the quality of the photoscreener in refractive ARFs detection and to find probably the best cut-off points. Results: In total, 1 986 children [mean age, (4.57±0.29) years] received comprehensive examinations, including 1 084 boys and 902 girls. The test ability of photoscreening was 99.04% (1 967/1 986) in the preschool children, and 96.56%(1 827/1 892) of the children got a reliable result within three screening attempts. In 538 children who had data of CR, refractive error of one child exceeded the upper limit of the photoscreener value setting, which was directly categorized as hyperopia, so in the end, 537 children were included to analyze the comparison between the two tests. The measurement values of photoscreening were lower than those of CR in sphere, cylinder and spherical equivalent [(0.75 (0.50, 1.25) D vs. 1.25 (1.00, 1.75) D, Z=-10.36, P<0.01; -0.50 (-0.75, -0.25) D vs. -0.25 (-0.75, 0.00) D, Z=-11.10, P<0.01; 0.63 (0.38, 0.88) D vs. 1.00 (0.75, 1.50) D, Z=-13.33, P<0.01]. The 95% limit of agreement cover rates between the photoscreening and CR in sphere, cylinder and spherical equivalent was 96.28% (517/537), 95.34% (512/537) and 96.65% (519/537), respectively. Based on the golden standard, 47 (8.74%) children had refractive ARFs, and the range of sensitivity, specificity, Youden index, positive predictive values and negative predictive values for detecting refractive ARFs of the 5 common used referral criteria was from 63.83% to 97.87%, from 53.36% to 97.56%, from 0.51 to 0.80, from 16.73% to 74.51% and from 96.57% to 99.62%, respectively. Considering particular refractive ARFs on the basis of the receiver operating characteristic curves, the optimal cut-off point for astigmatism was set at 1.38 D. Conclusion: Photoscreening could be an applicable tool to detect refractive ARFs in preschool children. (Chin J Ophthalmol, 2020, 56: 189-196).

[Analysis of follow-up results of chrono-chemotherapy or conventional chemotherapy combined with intensity modulated radiotherapy in locally advanced nasopharyngeal carcinoma].

Objective: To evaluate the long-term effect and safety of chrono-chemotherapy combined with intensity modulated radiotherapy (IMRT) in locally advanced nasopharyngeal carcinoma (NPC). Methods: 160 patients with locally advanced NPC were randomly divided into a chrono group and conventional group according to random number table. In the first stage, all patients underwent two cycles of induced chemotherapy, consisting of docetaxel, cisplatin and 5-Fu every 21 days. Notably, patients received chrono-moduated chemotherapy according to circadian rhythm in the chrono group, and conventional chemotherapy in the conventional group. Then, 21 days after the completion of first stage, three cycles of concurrent cisplatin chemotherapy every 21 days were given to all patients during IMRT. The median follow-up after the completion of radiotherapy was 31 months. Long-term side effects and the survival of patients were observed. Results: Patients in the chrono group had significantly lower rates of hearing loss (22.72%), dysphagia (0) and neck fibrosis (4.54%) compared with those in the conventional group (39.13%、8.69%, 15.94%, respectively, all P<0.05). Meanwhile, the 1- year overall survival rates (97.0% vs 92.8%), 3-year overall survival rates (80.3% vs 81.2%), 1-year progression free survival rates (95.5% vs 87.0%), 3-year progression free survival rates (71.2% vs 73.9%), 1-year locoregional relapse-free survival rates (97.0% vs 95.7%), 1-year locoregional relapse-free survival rates (92.4% vs 92.8%), 1-year distant metastasis-free survival rates (97.0% vs 98.6%) and 3-year distant metastasis-free survival rates (90.9% vs 91.3%) between the chrono group and the conventional group were not statistically significant (all P>0.05). Conclusions: Compared with conventional chemotherapy, chrono-chemotherapy combined with IMRT didn't affect long-term survival, but reducing the incidence of adverse events in patients with locally advanced NPC.

Clone, expression and plasminogen binding property of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis.

Fructose-1,6-bisphosphate aldolase (FbA), a well characterized glycometabolism enzyme, has been found to participate in other important processes besides the classic catalysis. To understand the important functions of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis (CsFbAs, CsFbA-1/2/3) in host-parasite interplay, the open reading frames of CsFbAs were cloned into pET30a (+) vector and the resulting recombinant plasmids were transformed into Escherichia coli BL21 (DE3) for expression of the proteins. Purified recombinant CsFbAs proteins (rCsFbAs) were approximately 45.0 kDa on 12% SDS-PAGE and could be probed with each rat anti-rCsFbAs sera by western blotting analysis. ELISA and ligand blot overlay indicated that rCsFbAs of 45.0 kDa as well as native CsFbAs of 39.5 kDa from total worm extracts and excretory-secretory products of Clonorchis sinensis (CsESPs) could bind to human plasminogen, and the binding could be efficiently inhibited by lysine analog ε-aminocaproic acid. Our results suggested that as both the components of CsESPs and the plasminogen binding proteins, three CsFbAs might be involved in preventing the formation of the blood clot so that Clonorchis sinensis could acquire enough nutrients from host tissue for their successful survival and colonization in the host. Our work will provide us with new information about the biological function of three CsFbAs and their roles in hostparasite interplay.

Circular RNA circHIPK3 serves as a prognostic marker to promote chronic myeloid leukemia progression.

Increasing evidence demonstrate that circular RNAs (circRNAs) play critical role in regulation of gene expression, which participate in the pathogenesis of cancer, including chronic myeloid leukemia (CML). In this study, we aimed to investigate the expression profiling of circHIPK3 in CML. We found that circHIPK3 was significantly upregulated in peripheral blood mononuclear cells (PBMC) and serum samples from CML compared with healthy controls. High circHIPK3 expression predicted a poor outcome of CML patients. Further loss-function experiments suggested the oncogenic role of circHIPK3 in CML. Our findings provide insights on the role of circHIPK3 in the development and treatment of CML.

Enantioconvergent hydrolysis of m-nitrostyrene oxide at an elevated concentration by Phaseolus vulgaris epoxide hydrolase in the organic/aqueous two-phase system.

(R)-m-Nitrophenyl-1,2-ethanediol (m-NPED) is a versatile and highly value-added chiral building block for the synthesis of some bioactive compounds, such as (R)-Nifenalol. To efficiently produce (R)-m-NPED through the enantioconvergent hydrolysis of racemic (rac-) m-nitrostyrene oxide (m-NSO) using the whole resting cells of Escherichia coli/pCold-pveh2 intracellularly expressing PvEH2, an epoxide hydrolase from Phaseolus vulgaris, two reaction systems were investigated. In the Na2 HPO4 -NaH2 PO4 buffer (50 mmol l-1 , pH 7·0) system, merely 15 mmol l-1 rac-m-NSO was successfully subjected to enantioconvergent hydrolysis, producing (R)-m-NPED with 86·0% enantiomeric excess (eep ) and 177·6 mg l-1  h-1 space-time yield (STY). The experimental result indicated that there is inhibitory effect of rac-m-NSO at high concentration on PvEH2. To efficiently increase the concentration of rac-m-NSO and the STY of (R)-m-NPED, petroleum ether was first selected to construct an organic/aqueous two-phase system. Then, both the volume ratio (vo /vb ) of petroleum ether to phosphate buffer and the weight ratio (wc /ws ) of E. coli/pCold-pveh2 dry cells to rac-m-NSO were optimized as 2 : 8 and 5 : 1, respectively. In the optimized petroleum ether/phosphate buffer two-phase system, the enantioconvergent hydrolysis of rac-m-NSO at 40 mmol l-1 (6·6 mg ml-1 ) was carried out at 25°C for 12 h using 33·0 mg ml-1 vacuum freeze-dried cells of E. coli/pCold-pveh2, producing (R)-m-NPED with 87·4% eep , 82·3% yield and 502·4 mg l-1  h-1 STY. SIGNIFICANCE AND IMPACT OF THE STUDY: Epoxide hydrolases play a crucial role in producing enantiopure epoxides and/or vicinal diols. However, numerous biocatalytic reactions of organic compounds, such as epoxides, in aqueous phase suffered various restrictions. Herein, the enantioconvergent hydrolysis of rac-m-NSO in two reaction systems was investigated using the whole cells of Escherichia coli/pCold-pveh2. As a result, the concentration of rac-m-NSO and the space-time yield of (R)-m-NPED in organic/aqueous two-phase system were significantly increased, when compared with those in aqueous phase. To our knowledge, this is the first report about the production of (R)-m-NPED from rac-m-NSO at an elevated concentration by PvEH2 in the two-phase system.

Clone, expression and plasminogen binding property of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis

@# Fructose-1,6-bisphosphate aldolase (FbA), a well characterized glycometabolism enzyme, has been found to participate in other important processes besides the classic catalysis. To understand the important functions of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis (CsFbAs, CsFbA-1/2/3) in host-parasite interplay, the open reading frames of CsFbAs were cloned into pET30a (+) vector and the resulting recombinant plasmids were transformed into Escherichia coli BL21 (DE3) for expression of the proteins. Purified recombinant CsFbAs proteins (rCsFbAs) were approximately 45.0 kDa on 12% SDS-PAGE and could be probed with each rat anti-rCsFbAs sera by western blotting analysis. ELISA and ligand blot overlay indicated that rCsFbAs of 45.0 kDa as well as native CsFbAs of 39.5 kDa from total worm extracts and excretory-secretory products of Clonorchis sinensis (CsESPs) could bind to human plasminogen, and the binding could be efficiently inhibited by lysine analog ε-aminocaproic acid. Our results suggested that as both the components of CsESPs and the plasminogen binding proteins, three CsFbAs might be involved in preventing the formation of the blood clot so that Clonorchis sinensis could acquire enough nutrients from host tissue for their successful survival and colonization in the host. Our work will provide us with new information about the biological function of three CsFbAs and their roles in hostparasite interplay

MicroRNA-383 suppresses pancreatic carcinoma development via inhibition of GAB1 expression.

OBJECTIVE: Pancreatic carcinoma (PC) is a serious malignancy associated with high morbidity and mortality rates. Previous studies have identified various microRNAs (miRNAs) involved in the development of PC; however, the role of miR-383 still remains unclear. This study investigates the role of miR-383 in the malignant transformation of PC. MATERIALS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to quantify miR-383 and Grb2 associated binding protein 1 (GAB1) RNA levels, and Western blot analysis was performed to measure protein expression. The ability of miR-383 to bind and regulate the expression of GAB1 was assessed using a Luciferase reporter assay. Cell Counting Kit-8 (CCK-8) experiments and flow cytometry analysis were used to assess cell proliferation and apoptosis, respectively. RESULTS: Down-regulation of miR-383 was associated with adverse clinical results and poor prognosis in PC patients. Mechanistically, miR-383 inhibited cell proliferation and promoted apoptosis of PANC-1 (human pancreatic cancer cell) cells. Our results show that miR-383 can act directly on GAB1 to inhibit its expression in PC. This downregulation of GAB1 limits cell proliferation and induced apoptosis of PANC-1 cells. CONCLUSIONS: MiR-383 suppresses tumor development and progression through the downregulation of GAB1 expression.