NLRP3 licenses NLRP11 for inflammasome activation in human macrophages.

Intracellular sensing of stress and danger signals initiates inflammatory innate immune responses by triggering inflammasome assembly, caspase-1 activation and pyroptotic cell death as well as the release of interleukin 1ß (IL-1ß), IL-18 and danger signals. NLRP3 broadly senses infectious patterns and sterile danger signals, resulting in the tightly coordinated and regulated assembly of the NLRP3 inflammasome, but the precise mechanisms are incompletely understood. Here, we identified NLRP11 as an essential component of the NLRP3 inflammasome in human macrophages. NLRP11 interacted with NLRP3 and ASC, and deletion of NLRP11 specifically prevented NLRP3 inflammasome activation by preventing inflammasome assembly, NLRP3 and ASC polymerization, caspase-1 activation, pyroptosis and cytokine release but did not affect other inflammasomes. Restored expression of NLRP11, but not NLRP11 lacking the PYRIN domain (PYD), restored inflammasome activation. NLRP11 was also necessary for inflammasome responses driven by NLRP3 mutations that cause cryopyrin-associated periodic syndrome (CAPS). Because NLRP11 is not expressed in mice, our observations emphasize the specific complexity of inflammasome regulation in humans.

ADAR1 mutation causes ZBP1-dependent immunopathology.

The RNA-editing enzyme ADAR1 is essential for the suppression of innate immune activation and pathology caused by aberrant recognition of self-RNA, a role it carries out by disrupting the duplex structure of endogenous double-stranded RNA species1,2. A point mutation in the sequence encoding the Z-DNA-binding domain (ZBD) of ADAR1 is associated with severe autoinflammatory disease3-5. ZBP1 is the only other ZBD-containing mammalian protein6, and its activation can trigger both cell death and transcriptional responses through the kinases RIPK1 and RIPK3, and the protease caspase 8 (refs. 7-9). Here we show that the pathology caused by alteration of the ZBD of ADAR1 is driven by activation of ZBP1. We found that ablation of ZBP1 fully rescued the overt pathology caused by ADAR1 alteration, without fully reversing the underlying inflammatory program caused by this alteration. Whereas loss of RIPK3 partially phenocopied the protective effects of ZBP1 ablation, combined deletion of caspase 8 and RIPK3, or of caspase 8 and MLKL, unexpectedly exacerbated the pathogenic effects of ADAR1 alteration. These findings indicate that ADAR1 is a negative regulator of sterile ZBP1 activation, and that ZBP1-dependent signalling underlies the autoinflammatory pathology caused by alteration of ADAR1.

Distinguishing Oligosaccharide Isomers Using Far-Infrared Ion Spectroscopy: Identification of Biomarkers for Inborn Errors of Metabolism.

Distinguishing isomeric saccharides poses a major challenge for analytical workflows based on (liquid chromatography) mass spectrometry (LC-MS). In recent years, many studies have proposed infrared ion spectroscopy as a possible solution as the orthogonal, spectroscopic characterization of mass-selected ions can often distinguish isomeric species that remain unresolved using conventional MS. However, the high conformational flexibility and extensive hydrogen bonding in saccharides cause their room-temperature fingerprint infrared spectra to have broad features that often lack diagnostic value. Here, we show that room-temperature infrared spectra of ion-complexed saccharides recorded in the previously unexplored far-infrared wavelength range (300-1000 cm-1) provide well-resolved and highly diagnostic features. We show that this enables distinction of isomeric saccharides that differ either by their composition of monosaccharide units and/or the orientation of their glycosidic linkages. We demonstrate the utility of this approach from single monosaccharides up to isomeric tetrasaccharides differing only by the configuration of a single glycosidic linkage. Furthermore, through hyphenation with hydrophilic interaction liquid chromatography, we identify oligosaccharide biomarkers in patient body fluid samples, demonstrating a generalized and highly sensitive MS-based method for the identification of saccharides found in complex sample matrices.

[Definition of severe pulmonary tuberculosis: a scoping review].

Objective: To clarify the definition of severe pulmonary tuberculosis and its inclusion criteria by summarizing and analyzing the studies of severe pulmonary tuberculosis (TB). Methods: A systematic search of Medline (via PubMed), Cochrane Library, Web of Science, Web of Science, Epistemonikos, Embase, CNKI, WanFang database, and CBM database was conducted to collect studies published between 2017 and 2022 on patients with severe pulmonary TB. Searches were performed using a combination of subject terms and free words. The search terms included: tuberculosis, severe, serious, intensive care, critical care, respiratory failure, mechanical ventilation, hospitalization, respiratory distress syndrome, multiple organ failure, pulmonary heart disease, and pneumothorax. The definitions and inclusion criteria for severe pulmonary TB in the included studies were extracted. Results: A total of 19 981 studies were identified and 100 studies were finally included, involving 8 309 patients with severe pulmonary TB. A total of 8 (8.00%) studies explicitly mentioned the definition of severe pulmonary TB, and 53 (53.00%) studies clearly defined the inclusion criteria for patients with severe pulmonary TB. A total of 5 definitions and 30 inclusion criteria were extracted. A total of 132 dichotomous variables and 113 continuous variables were included in the outcome indicators related to patients with severe pulmonary TB of concern in the studies. Conclusions: The definition and diagnostic criteria for severe TB are unclear, and there is an urgent need to develop a clear definition and diagnostic criteria to guide clinical practice.

[Determination of 4-methyl-2-pentanol in workplace air by solvent desorption-gas chromatography].

Objective: To establish a method for the determination of 4-methyl-2-pentanol in the air of workplace by gas chromatography. Methods: In January 2022, 4-methyl-2-pentanol in the air of workplace was collected by activated carbontube, eluted with dichloromethane-methanol (95∶5, V/V), separated by capillary column and determined by gas chromatogram. Results: The limit of detection for 4-methyl-2-pentanol was 0.04 µg/ml. The linear range of 4-methyl-2-pentanol was 0.16-1616.60 µg/ml, with the regression equation of y=1.94x-5.48, and the coefficient correlation was 0.99958, and the minimum detection concentration was 0.03 mg/m(3) (collected sample volume was 1.50 L). The within-run precisions were 1.08%-1.75% and the between-run precisions were 1.41%-2.52%. The desorption rates were 95.15%-99.91%. The samples could be stored at least 3 days at room temperature and 7 days at 4 ℃ without significant loss. Conclusion: The method has the advantages of good precision, high sensitivity and simple operation. It is suitable for the determination of 4-methyl-2-pentanol in the air of workplace.

Transcriptome profiling reveals morphogenesis-related candidate genes and pathways in the chick embryonic small intestine.

1. A better understanding of intestinal development is essential for the intestinal health of poultry. Intestinal villification starts on embryo day E15 and is generally completed before hatching (E21). The development of lymphoid organs in the intestine starts during embryogenesis. However, transcriptional information on the processing of intestinal morphogenesis and immune development during chick embryogenesis is limited.2. In this work, RNA-sequencing was performed using 12 biological replicates to investigate Hy-Line brown chick embryonic small intestinal transcription at E15 and E21. Differentially expressed genes (DEGs) between E15 and E21 were identified. GO and KEGG enrichment analyses, based on the DEGs, were performed to identify key GO terms in the biological process category and key KEGG pathways. PPI networks were constructed based on the DEGs in the key pathways to screen hub genes. The embryonic small intestinal morphology and IgA distribution were observed by histological processing. The serum levels of IgA and lysozyme were measured by ELISA.3. A total of 76.38 Gb of high-quality RNA-sequencing data were generated and uploaded. A total of 2,676 DEGs, between E15 and E21, were identified. Structural development and villification of the small intestine at E15 tended to proceed via the expression of nervous system development-related genes. A combination of the histological and serological results with the transcriptome data indicated that the identified genes and pathways may be strong candidates for intestinal morphogenesis-regulation.4. The small intestine appears to have developed a relatively complete morphology and transport, metabolism, digestion and immunity functions by E21. This work provided a transcriptome profile of the chick embryonic small intestine and provided insights into the intestinal development and health of poultry.

[A follow-up study on the introduction of vegetables and fruits during infants aged 4-8 months in Beijing and Chenzhou City of Hunan Province].

OBJECTIVE: To investigate the introduction of vegetables and fruits in 4-8 months old infants, and to describe the maternal and infants' characteristics associated with the introduction of vegetables and fruits. METHODS: Mother-infant dyads (n=228) were recruited from 12 to 16 weeks postpartum and formally entered the study at 4 months of age. Data collected via face to face interview at 4-8 months postpartum, including the timing and types of added vegetables and fruits, as well as a variety of maternal and infant characteristics (n=204). Rank sum test and multiple linear regression were used to analyze the maternal and infant characteristics related to the introduction of vegetables and fruits. RESULTS: The time of introducing vegetables was concentrated at the age of 7 months, and the time of adding fruits was mainly at 6 months. Fruits were added earlier than vegetables (P < 0.001), and the variety of the added fruits was higher than that of vegetables (P=0.045). 48% (n=98) of infants had no more than three types of fruits and vegetables at 8 months. Only 9.8% (n=20) had added more than 10 kinds of fruits and vegetables at 8 months. Green leafy vegetables were the most commonly added vegetable, and apple was the most popular fruit. Compared with women who were 35 years of age or younger, women beyond 35 years old introduced vegetables to their babies 0.6 months later. 4-month-old exclusively breastfed infants had vegetables 0.4 months later than mixed-fed infants. Women with a bachelor's degree or above added 2-3 more types of fruits and vegetables to their babies than those with junior high school education and below. CONCLUSION: The adding time of fruits was earlier than that of vegetable. Apples and green leafy vegetables are commonly added. Women with lower educational backgrounds add fewer types of fruits and vegetables to their babies. Mothers who choose exclusive breastfeeding and those over 35 years of age at childbirth add vegetables to their babies later than others. They should be targeted for health promotion programs that aim to improve the intake of fruits and vegetables among infants.

Development of the Asia Pacific Consortium on Osteoporosis (APCO) Framework: clinical standards of care for the screening, diagnosis, and management of osteoporosis in the Asia-Pacific region.

Guidelines for doctors managing osteoporosis in the Asia-Pacific region vary widely. We compared 18 guidelines for similarities and differences in five key areas. We then used a structured consensus process to develop clinical standards of care for the diagnosis and management of osteoporosis and for improving the quality of care. PURPOSE: Minimum clinical standards for assessment and management of osteoporosis are needed in the Asia-Pacific (AP) region to inform clinical practice guidelines (CPGs) and to improve osteoporosis care. We present the framework of these clinical standards and describe its development. METHODS: We conducted a structured comparative analysis of existing CPGs in the AP region using a "5IQ" model (identification, investigation, information, intervention, integration, and quality). One-hundred data elements were extracted from each guideline. We then employed a four-round Delphi consensus process to structure the framework, identify key components of guidance, and develop clinical care standards. RESULTS: Eighteen guidelines were included. The 5IQ analysis demonstrated marked heterogeneity, notably in guidance on risk factors, the use of biochemical markers, self-care information for patients, indications for osteoporosis treatment, use of fracture risk assessment tools, and protocols for monitoring treatment. There was minimal guidance on long-term management plans or on strategies and systems for clinical quality improvement. Twenty-nine APCO members participated in the Delphi process, resulting in consensus on 16 clinical standards, with levels of attainment defined for those on identification and investigation of fragility fractures, vertebral fracture assessment, and inclusion of quality metrics in guidelines. CONCLUSION: The 5IQ analysis confirmed previous anecdotal observations of marked heterogeneity of osteoporosis clinical guidelines in the AP region. The Framework provides practical, clear, and feasible recommendations for osteoporosis care and can be adapted for use in other such vastly diverse regions. Implementation of the standards is expected to significantly lessen the global burden of osteoporosis.

Monoclonal antibodies and chimeric antigen receptor (CAR) T cells in the treatment of colorectal cancer.

Colorectal cancer (CRC) is the third most common cancer and the second leading cause of cancer deaths worldwide. Besides common therapeutic approaches, such as surgery, chemotherapy, and radiotherapy, novel therapeutic approaches, including immunotherapy, have been an advent in CRC treatment. The immunotherapy approaches try to elicit patients` immune responses against tumor cells to eradicate the tumor. Monoclonal antibodies (mAbs) and chimeric antigen receptor (CAR) T cells are two branches of cancer immunotherapy. MAbs demonstrate the great ability to completely recognize cancer cell-surface receptors and blockade proliferative or inhibitory pathways. On the other hand, T cell activation by genetically engineered CAR receptor via the TCR/CD3 and costimulatory domains can induce potent immune responses against specific tumor-associated antigens (TAAs). Both of these approaches have beneficial anti-tumor effects on CRC. Herein, we review the different mAbs against various pathways and their applications in clinical trials, the different types of CAR-T cells, various specific CAR-T cells against TAAs, and their clinical use in CRC treatment.

[Anatomical study and clinical application of in situ reduction and fixation of anterior medial fenestration approach of femoral head fracture].

Objectives: To explore the feasibility of anterior medial fenestration approach in situ reduction and fixation in the treatment of PipkinⅠ and Ⅱ femoral head fractures,and to explore the clinical effect of this operation. Methods: Hips of two anti-corrosion adult specimens treated with formalin were dissected, then anatomical structures and directional characteristics of anterior medial main muscles,ligaments,blood vessels and nerves were observed.The anterior medial fenestration approach was performed on bilateral hips of four fresh frozen specimens to determine pulling direction of stripped muscles and ligaments required during operation,and to observe and analyze vascular and nerve traction protection directions exposed in the approach.Determine extent of exposure to the approach and assess feasibility of this approach.The clinical data of 12 patients with Pipkin Ⅰ and Ⅱ femoral head fractures who underwent in situ reduction and fixation of anterior medial fenestration at Department of Orthopaedics,Affiliated Hospital of Chengdu University from February 2016 to April 2018 were retrospectively analyzed.There were 3 males and 9 females with an age of 48.5 years(range:37 to 59 years).There were 8 cases of Pipkin type Ⅰ and 4 cases of Pipkin type Ⅱ.The operation time,blood loss,fracture healing time,last Thompson-Epstein evaluation and Harris score were observed. Results: Anterior medial fenestration approach to expose the femoral head in 4 bilateral hips with a total of 8 sides of fresh frozen specimens.The upper boundary of observation fenestration was pubic body (anterior acetabulum),and the outer upper boundary was iliacus and psoas muscle.The lateral boundary is rectus femoris and femoral vessels,the lower boundary was transverse branch of the medial femoral circumflex artery and vein.The medial boundary was pubis muscle,short adductor muscle and long adductor muscle.Pubofemoral and iliofemoral ligament were seen in fenestration. Four quadrants in front of femoral head in fenestration can be seen after cutting switch capsule active hip joint. In 12 patients with femoral head fracture,the operation time was 107.5 minutes(range:90 to 135 minutes),and the intraoperative bleeding volume was 115.0 ml(range:85 to 150 ml).The patients were followed up for 18.6 months(range:12 to 28 months).The fracture healing time of 12 patients was 144.2 days(range:120 to 180 days).The curative effect was evaluated according to Thompson-Epstein standard at the last follow-up:excellent in 6 cases,good in 4 cases and fair in 2 cases.At the last follow-up,the Harris score of hip joints was 85.1(range:75 to 93). Conclusions: Anterior medial fenestration in situ reduction and fixation surgery is feasible for the treatment of Pipkin Ⅰ and Ⅱ femoral head fractures. The short and midterm follow-up reveal satisfactory effect.

[Study on the correlation between dietary N-glycolylneuraminic acid intake and chronic inflammation state of body].

Objective: To explore the correlation between dietary N-glycolylneuraminic acid (Neu5Gc) intake and chronic inflammation state of body. Methods: A total of 306 samples of 102 types of food were purchased from a supermarket in Xiamen in September 2019, including grains, meat, poultry, seafood, eggs, beans, dairy products, vegetables and fruits. The content of Neu5Gc in food was determined by liquid chromatography-mass spectrometry. A total of 500 healthy freshmen from Xiamen University were selected by using a simple random sampling method. The food frequency questionnaire was used to investigate the food intake in the past year. The food intake was corrected by 3 consecutive 24-hour recalls, and the amount of Neu5Gc intake was calculated. The concentration of anti-Neu5Gc antibody, C-reactive protein (CRP) and interleukin-6 (IL-6) in serum was detected. Spearman correlation analysis was used to explore the correlation between Neu5Gc intake and anti-Neu5Gc antibody, CRP and IL-6 levels. Results: Neu5Gc was mainly found in red meat and liquid dairy products. The contents of Neu5Gc in beef, lamb and pork were (30.32±2.84), (20.39±4.73) and (5.58±1.04) mg/kg, respectively, and in liquid milk and yogurt were (10.87±1.54) and (6.91±0.24) mg/L, respectively. The M (P25, P75) intake of Neu5Gc for all participants was 4.62 (2.20, 8.60) mg/d. The M(P25, P75) intake of Neu5Gc for males about 6.60(2.83, 10.20) was higher than that for females about [3.84 (1.84, 6.35) mg/d] (P<0.001). The M (P25, P75) of serum anti-Neu5Gc, CRP and IL-6 levels were 3.07 (2.17, 4.14) µg/ml, 0.37 (0.22, 0.87) mg/ml and 61.82 (12.23, 315.30) pg/ml, respectively. Spearman correlation analysis showed that the intake level of Neu5Gc was positively correlated with serum anti-Neu5Gc antibody, CRP and IL-6 levels, with rs values about 0.222, 0.102 and 0.126, respectively (all P values <0.05). Conclusion: Dietary Neu5Gc intake is mainly from red meat and liquid dairy products, and its intake level is positively correlated with chronic inflammatory state of body.

[Reduction of the atlantoaxial dislocation associated with basilar invagination through single-stage posterior approach: using Xuanwu occipital-cervical reduction surgical suite].

Objective: To examine the effect of posterior reduction in atlantoaxial dislocation (AAD) associated with basilar invagination(BI) using Xuanwu occipital-cervical fusion system in single stage. Methods: Thirty-seven AAD accompanied with BI cases treated at Department of Neurosurgery, Xuanwu Hospital, Capital Medical Universiy and the Second Hospital of Hebei Medical University were retrospective analyzed. There were 15 males and 22 females with age of (42.3±12.3)years (range: 18-69 yars). All the cases had congenital osseous abnormalities, such as assimilation of atlas and abnormal cervical fusion. Anterior tissue was released through posterior route followed by cage implantation into facet joint and occipital-cervical fixation with cantilever technique. The clinical results were evaluated using Japanese Orthopedic Association scale(JOA) and the main radiological measurements including anterior atlantodental interval (ADI),the distance of odontoid tip above Chamberlain line,clivus-canal angle (CCA) and the length of syrinx were collected.The preoperative and postoperative JOA score and radiological measurements were compared by paired t-test. Results: The mean JOA score of the patients increased from 10.5 to 14.4 at the one-year follow-up(t=14.3,P=0.00).Complete reduction of AAD and BI was achieved in 34 patients.The mean clivus-canal angle improved from 118.0 degrees preoperative to 143.7 degrees postoperative(t=6.2,P=0.00). Shrinkage of the syrinx was observed 1 week after surgery in 24 patients, and 6 months in 31 patients. Twenty-eight patients achieved bone fusion 6 months after surgery. All the patients achieved bone fusion 12 months after surgery. One-side vertebral artery occlusion was diagnosed in 1 case postoperatively for transient dizziness, and relieved in 2 weeks. Two patients developed moderate neck pain after surgery, and relieved in 1 month. No implant failure, spacer subsidence or infection was observed. Conclusions: The treatment of AAD associated with BI using Xuanwu occipital-cervical fusion system from posterior approach in single stage is effective and safe. Cage implantation intraarticularly and fixation with cantilever technique achieve complete reduction in most cases.

[Comparison of external fixation with or without limited internal fixation for open knee fractures].

Objective: To explore the characteristics and methods of different fixation methods and prevention of open knee joint fracture. Methods: The data of 86 cases of open knee joint fracture admitted from January 2002 to December 2015 in Department of Orthopaedics, Affiliated Hospital of Chengdu University were analyzed retrospectively.There were 65 males and 21 females aged of 38.6 years. There were 38 cases treated with trans articular external fixation alone, 48 cases were in the trans articular external fixation plus auxiliary limited internal fixation group. All the patients were treated according to the same three stages except for different fixation methods. Observation of external fixation and fracture fixation, fracture healing, wound healing and treatment, treatment and related factors of infection control and knee function recovery. χ(2) test was used to analyze data. Results: Eleven patients had primary wound healing, accounting for 12.8%. Seventy-five patients had two wounds healed, accounting for 87.2%. Only 38 cases of trans articular external fixator group had 31 cases of articular surface reduction, accounting for 81.6%; Five cases of trans articular external fixator assisted limited internal fixation group had 5 cases of poor reduction, accounting for 10.4%; There was significant difference between the two groups (χ(2)=44.132, P<0.05). Take a single cross joint external fixation group, a total of 23 cases of patients with infection, accounted for 60.5% of external fixation group; trans articular external fixation assisted limited internal fixation group there were 30 cases of patients with infection, accounting for the assistance of external fixator and limited internal fixation group 62.5%; There was significant difference between the two groups(χ(2)=0.035, P>0.05). Five cases of fracture nonunion cases of serious infection, patients voluntarily underwent amputation. The Lysholm Knee Scale: In the external fixation group, 23 cases were less than 50 points, accounting for 60.5%, 15 cases were more than 50 points, accounting for 39.5%, external fixation and limited internal fixation group 20 cases were less than 50 points, accounting for 41.7%, 28 cases were more than 50 points, accounting for 58.3%; There was significant difference between the two groups(χ(2)=1.279, P>0.05). Conclusions: Prevention and control of infection is a central link in the treatment of open fracture of the knee. Trans articular external fixator plus limited internal fixation is an important measure to treat open fracture of the knee-joint.

[Research of the biomarkers in pulmonary and extrapulmonary acute respiratory distress syndrome].

Objective: To explore the biomarkers that applied to pulmonary acute respiratory distress syndrome (ARDS) and extrapulmonary ARDS respectively. Methods: A total of 49 eligible patients with ARDS were selected from Department of Respiratory intensive care unite (ICU) , Department of General ICU and Department of Emergency ICU, the Second Affiliated Hospital of Nanchang University, during April 2015 and February 2016, and divided into pulmonary ARDS group (n=26) and extrapulmonary ARDS group (n=23) according to primary causes. Totally 19 cases from healthy examination personnel were served as control. Acute physiology and chronic health evaluation Ⅲ (APACHE Ⅲ) score and lung injury score of all hospitalized patients were completed within 24 hours after the diagnosis of ARDS and the serum levels of angiogenin2 (Ang-2) , surfactant protein D (SP-D), soluble receptor for advanced glycation end products (sRAGE), Krebs von den Lungen-6 (KL-6) were measured. Then the 28-day outcomes were observed. The serum levels of the biomarkers were compared between each group and correlation analysis was conducted between different indexes. Finally, The relationship between the four biomarkers and the outcome of each group was analyzed. Results: The serum level of Ang-2 in pulmonary ARDS group and extrapulmonary ARDS group was significantly higher than that in control group[1 462 (569, 3 638) and 4 568 (2 236, 8 457) vs 460 (389, 578) ng/L, all P<0.05], and the serum level of Ang-2 was significantly higher in extrapulmonary ARDS group than that in pulmonary ARDS group (P<0.05). The serum level of SP-D in pulmonary ARDS group and extrapulmonary ARDS group was significantly higher than that in control group[5.9 (2.5, 22.7) mg/L and 3.7 (1.7, 6.5) vs 1.9 (0.6, 4.4) mg/L, all P<0.05], and the serum level of SP-D was significantly higher in pulmonary ARDS group than that in extrapulmonary ARDS group (P<0.05). The serum level of sRAGE in pulmonary ARDS group and extrapulmonary ARDS group was significantly higher than that in control group[238 (101, 583) and 229 (114, 443) vs 117 (88, 176) ng/L, all P<0.05]. The serum level of KL-6 in pulmonary ARDS group was significantly higher than that in control group[10 435 (3 401, 17 607) vs 4 780 (3 962, 5 320) U/L, P<0.05]. Increased serum levels of SP-D (OR=1.089, 95% CI: 1.001-1.085) and KL-6 (OR=1.206, 95% CI: 1.016-1.432) at the early stage were the independent prognostic factors for the 28-day mortality for pulmonary ARDS. Advanced age (OR=2.622, 95% CI: 1.046-6.574) was the independent prognostic factor for the 28-day mortality for extrapulmonary ARDS. Conclusion: SP-D and KL-6 may be the serum biomarkers of pulmonary ARDS and Ang-2 may be the serum biomarker of extrapulmonary ARDS.

[Clinical outcome of totally thoracoscopic cardiac surgery for mitral valve replacement: a series of 634 cases].

OBJECTIVE: To summarize the clinical outcome of totally thoracoscopic cardiac surgery for mitral valve replacement. METHODS: Clinical data of 634 cases undergoing totally thoracoscopic cardiac surgery for mitral valve replacement from May 2004 to February 2016 in Department of Thoracoscopic Cardiacsurgery, Shanghai Yodak Cardiothoracic Hospital was analyzed retrospectively. There were 292 male and 342 female patients, aged from 17 to 68 years with a mean of (45±13) years. All the 634 patients had moderate-severe mitral valve stenosis and (or) incompetence, 263 patients had moderate-severe tricuspid valve incompetence, 356 patients had atrial fibrillation, 46 patients had left atrium thrombosis. Cardiopulmonary bypass was established with right femoral artery and a single 2 stage venus cannula in the right atrium. The ascending aorta was cross-clamped and the myocardium was protected by coronary perfusion with cold crystalloid cardioplegia. Totally thoracoscopic mitral valve replacement were performed. RESULTS: Thirteen cases had incision expanded and 8 cases had conversions to sternotomy. Cardiopulmonary bypass and aortic cross-clamp time were (89±18) minutes and (51±12) minutes, respectively. Operation time was (3.1±1.2) hours. Mechanical ventilation time and intensive care unit stay were (17±6) hours and (27±8) hours, respectively. Postoperation drainage quantity was (390±70) ml. The hospital days was (9.2±2.1) days. There were 5 cases in-hospital deaths. Postoperative complications occurred in 42 cases (6.6%), including 18 cases of right hemoneumothorax, 12 cases of reoperation for bleeding, 3 cases of perivalvular leakage (reoperation was done in 1 patient), 3 cases of low cardiac output syndrome, 2 cases of acute renal failure, 2 cases of inferior vena cava injury, 1 case of right femoral artery thrombosis and liver injury, respectively. The mean duration of follow-up was (58±9) months in 608 cases, with a follow-up rate of 96.7% (608/629). Three patients had died during the period of follow-up caused by congestive heart failure (2 patients) and stroke (1 patient). Late complication among 605 survivors were 37 cases, including 32 cases of moderate tricuspid valve insufficiency, 3 cases of stroke, 1 case of perivalvular leakage and infective endocarditis, respectively.There was no reoperation during the period of follow-up. CONCLUSION: Totally thoracoscopic cardiac surgery for mitral valve replacement is safe and effective, with unique superiority and clinical feasible.

Commercial-scale biotherapeutics manufacturing facility for plant-made pharmaceuticals.

Rapid, large-scale manufacture of medical countermeasures can be uniquely met by the plant-made-pharmaceutical platform technology. As a participant in the Defense Advanced Research Projects Agency (DARPA) Blue Angel project, the Caliber Biotherapeutics facility was designed, constructed, commissioned and released a therapeutic target (H1N1 influenza subunit vaccine) in <18 months from groundbreaking. As of 2015, this facility was one of the world's largest plant-based manufacturing facilities, with the capacity to process over 3500 kg of plant biomass per week in an automated multilevel growing environment using proprietary LED lighting. The facility can commission additional plant grow rooms that are already built to double this capacity. In addition to the commercial-scale manufacturing facility, a pilot production facility was designed based on the large-scale manufacturing specifications as a way to integrate product development and technology transfer. The primary research, development and manufacturing system employs vacuum-infiltrated Nicotiana benthamiana plants grown in a fully contained, hydroponic system for transient expression of recombinant proteins. This expression platform has been linked to a downstream process system, analytical characterization, and assessment of biological activity. This integrated approach has demonstrated rapid, high-quality production of therapeutic monoclonal antibody targets, including a panel of rituximab biosimilar/biobetter molecules and antiviral antibodies against influenza and dengue fever.

Molecular and biochemical characterization of recombinant guinea pig tumor necrosis factor-alpha.

Tumor necrosis factor alpha (TNF-α) is a cytokine which plays opposing roles in the context of infectious disease pathogenesis. TNF-α is essential for the development of a protective immune response to some pathogens, for example, Mycobacterium tuberculosis, by synergizing with other cytokines. However, exorbitant or uncontrolled TNF-α activity may also drive pathology and disease symptoms in many infectious diseases. In order to elucidate the beneficial and detrimental roles of TNF-α in tuberculosis (TB) and other diseases for which the guinea pig is the small animal model of choice, recombinant guinea pig (rgp)TNF-α has been produced using prokaryotic expression systems. However, it is unknown whether posttranslational modifications which cannot be made in the prokaryotic expression systems may be important for rgpTNF-α structure and function. Therefore, we carried out a comparative study by expressing rgpTNF-α in prokaryotic and eukaryotic expression systems and analyzed the eukaryotic-expressed rgpTNF-α for the presence of posttranslational modifications by subjecting it to NanoLC-MS/MS. We conclude that the eukaryotic-expressed rgpTNF-α lacks posttranslational modifications, and we found no significant difference in terms of the biological activity between prokaryotic- and eukaryotic-expressed rgpTNF-α. Taken together, results from our study show that a prokaryotic expression system can be used for generating large amounts of rgpTNF-α without concern for the biological integrity.

Characterization of OsPM19L1 encoding an AWPM-19-like family protein that is dramatically induced by osmotic stress in rice.

The plant-specific AWPM-19-domain proteins play important roles in plant development and stress responses. In the current study, OsPM19L1 encoding Oryza sativa AWPM-19-like protein 1 was isolated from rice. Tissue-specific gene expression analysis revealed that OsPM19L1 was highly expressed in the leaf sheath of rice. Interestingly, expression of OsPM19L1 was high at the early stage of panicle development and decreased thereafter. qRT-PCR analysis indicated that OsPM19L1 was dramatically induced by 20% PEG stress (>600-fold), exogenous abscisic acid (>350-fold), salt and cold stress. Subcellular localization assay suggested that the OsPM19L1-GFP (green fluorescent protein) fusion protein was localized in the membrane system in rice cells. Moreover, under stress conditions, OsPM19L1 expression was enhanced in an ABI5-Like1 (ABL1) deficiency rice mutant, abl1, suggesting that ABL1 negatively regulates OsPM19L1 gene expression. Thus, OsPM19L1 appears to be closely associated with stress tolerance through ABA-dependent pathway in rice.