Objective:To study the clinical characteristics and risk factors of nephrocalcinosis in preterm infants.Methods:From March 2021 to August 2021, all preterm infants admitted to NICU of our hospital were retrospectively analyzed. The infants were assigned into nephrocalcinosis group and non-nephrocalcinosis group according to urinary tract ultrasound. Clinical data including gestational age, birth weight(BW), nutritional support strategy and complications were reviewed.Results:A total of 40 preterm infants (<34 weeks) were enrolled. 9 cases were in the nephrocalcinosis group and 31 cases in the non-nephrocalcinosis group. The nephrocalcinosis group had lower BW[(1 167±214) g vs.(1 586±215) g], higher calcium [6.9 (5.1, 8.7) g vs.3.3 (2.1, 6.8) g] and vitamin D intake [3.2(2.5, 4.2)×10 4U vs.1.7(1.1, 3.2)×10 4U] during hospitalization. No significant differences existed between the two groups on the following items:blood calcium and phosphate, 25-hydroxyvitamin D, feeding strategy, time to reach full enteral feeding(TFF), furosemide dosage and respiratory support duration ( P>0.05). In the nephrocalcinosis group, the median age of diagnosing nephrocalcinosis was 40.0(30.0, 52.5)d after birth. 5 cases showed bilateral nephrocalcinosis. 5 cases in the nephrocalcinosis group received renal tubule function examination,4 cases had increased urine β2 microglobulin and 2 cases had increased urine α1 microglobulin. 7 cases had elevated urine calcium in the nephrocalcinosis group. Follow-up showed that nephrocalcinosis disappeared 3-9 months after birth. Conclusions:BW, total calcium and vitamin D intake are risk factors for nephrocalcinosis in preterm infants. Increased urine β2 microglobulin and calcium levels are common co-morbidities in preterm infants with nephrocalcinosis.

Objective To construct c-Met CAR-T cells secreting PD-1 antibodies to reduce immune inhibitory effect of tumor cells and enhance the efficacy of CAR-T cell therapy against pancreatic cancer.Methods Kaplan-Meier Plotter,GEPIA,and Timer 2.0 bioinformatics databases were used to analyze c-Met expression in pancreatic cancer and its correlation with survival and immune infiltration status.In clinical samples of pancreatic cancer and pancreatic cancer Aspc-1 cells,c-Met and PD-L1 expressions were detected using immunohistochemistry or flow cytometry.Using gene editing technology,PD-1 secretory antibodies and HIS tags were linked to second-generation c-Met CAR molecules to construct PD-1/c-Met CAR plasmids,which were then packaged into lentiviruses for infection of activated T cells.The positive rate and cell subset distribution of CAR-T cells were analyzed with flow cytometry,and secretory PD-1 antibodies in cell supernatants were detected using Western blotting.The target cell killing efficiency and proliferative activity of the modified CAR-T cells were evaluated after activation,and cytokine secretion was analyzed using ELISA.Results The expression of c-Met was significantly higher in pancreatic cancer than in normal tissues,and its expression level was negatively correlated with the patients'survival and positively correlated with immune cell infiltration.The clinical samples of pancreatic cancer tissues expressed significantly higher levels of c-Met and PD-L1 than the adjacent tissues,and 90.7%and 57.7%of Aspc-1 cells were positive for c-Met and PD-L1,respectively.The constructed PD-1/c-Met CAR-T cells were capable of secreting PD-1 antibodies and showed a significantly higher killing efficiency against tumor cells than c-Met CAR-T cells at an effector-to-target ratio of 20:1,with also a higher proliferative activity after target cell stimulation and higher levels of IL-2 and TNF-α secretin.Conclusion PD-1/c-Met CAR-T cells have higher killing efficiency against pancreatic cancer cells with also higher proliferative activity than c-Met CAR-T cells.

Objective To construct c-Met CAR-T cells secreting PD-1 antibodies to reduce immune inhibitory effect of tumor cells and enhance the efficacy of CAR-T cell therapy against pancreatic cancer.Methods Kaplan-Meier Plotter,GEPIA,and Timer 2.0 bioinformatics databases were used to analyze c-Met expression in pancreatic cancer and its correlation with survival and immune infiltration status.In clinical samples of pancreatic cancer and pancreatic cancer Aspc-1 cells,c-Met and PD-L1 expressions were detected using immunohistochemistry or flow cytometry.Using gene editing technology,PD-1 secretory antibodies and HIS tags were linked to second-generation c-Met CAR molecules to construct PD-1/c-Met CAR plasmids,which were then packaged into lentiviruses for infection of activated T cells.The positive rate and cell subset distribution of CAR-T cells were analyzed with flow cytometry,and secretory PD-1 antibodies in cell supernatants were detected using Western blotting.The target cell killing efficiency and proliferative activity of the modified CAR-T cells were evaluated after activation,and cytokine secretion was analyzed using ELISA.Results The expression of c-Met was significantly higher in pancreatic cancer than in normal tissues,and its expression level was negatively correlated with the patients'survival and positively correlated with immune cell infiltration.The clinical samples of pancreatic cancer tissues expressed significantly higher levels of c-Met and PD-L1 than the adjacent tissues,and 90.7%and 57.7%of Aspc-1 cells were positive for c-Met and PD-L1,respectively.The constructed PD-1/c-Met CAR-T cells were capable of secreting PD-1 antibodies and showed a significantly higher killing efficiency against tumor cells than c-Met CAR-T cells at an effector-to-target ratio of 20:1,with also a higher proliferative activity after target cell stimulation and higher levels of IL-2 and TNF-α secretin.Conclusion PD-1/c-Met CAR-T cells have higher killing efficiency against pancreatic cancer cells with also higher proliferative activity than c-Met CAR-T cells.

Objective:To explore the association between grip strength and cognitive function in elderly people aged 65 years and over.Methods:Information on grip strength, cognitive function, and lifestyle in the elderly population aged 65 years and over in Wuhan was collected by unified professionally trained investigators.A total of 533 study subjects aged(70.7±5.1)years were grouped by quartile into four grip strength groups of Q1(<18.6 kg), Q2(18.6~24.1 kg), Q3(24.2~31.1 kg), Q4(>31.1 kg).Multiple linear regression and Logistic regression analyses were used to analyze the relationship of grip strength with cognitive function scores and cognitive impairment.Results:The mean grip strength of the 533 subjects was(24.94±9.15)kg.After adjusting for sociodemographic characteristics, lifestyle factors, history of diseases, as compared with grip strength Q1 group, the linear regression coefficients(Beta value)of cognitive scores and 95% confidence intervals(95% CI)showed statistically significantly positive correlation[0.45(-0.36, 1.26)、0.40(-0.52, 1.32)and 1.19(0.07, 2.31), all P<0.05]only between cognitive scores and grip strength Q2、Q3 and Q4 value; and the odds ratio and 95% CI of incidence of cognitive impairment were 0.97(0.43, 2.21)for grip strength Q2, 0.79(0.30, 2.06)for grip strength Q3, and 0.22(0.05, 0.92)for grip strength Q4.Considering grip strength as the continuous variable, the risk of cognitive impairment was decreased by 6% and the cognitive score was increased by 0.07 with per 1kg increase of grip strength. Conclusions:The results of this study suggest that there is a positive correlation between grip strength and cognitive function in the elderly population, and a lower grip strength is related to increased risk of cognitive impairment.More attention should be paid to the grip strength of the elderly in the community.

New threats posed by the emerging circulating variants of SARS-CoV-2 highlight the need to find conserved neutralizing epitopes for therapeutic antibodies and efficient vaccine design. Here, we identified a receptor-binding domain (RBD)-binding antibody, XG014, which potently neutralizes β-coronavirus lineage B (β-CoV-B), including SARS-CoV-2, its circulating variants, SARS-CoV and bat SARSr-CoV WIV1. Interestingly, antibody family members competing with XG014 binding show reduced levels of cross-reactivity and induce antibody-dependent SARS-CoV-2 spike (S) protein-mediated cell-cell fusion, suggesting a unique mode of recognition by XG014. Structural analyses reveal that XG014 recognizes a conserved epitope outside the ACE2 binding site and completely locks RBD in the non-functional "down" conformation, while its family member XG005 directly competes with ACE2 binding and position the RBD "up". Single administration of XG014 is effective in protection against and therapy of SARS-CoV-2 infection in vivo. Our findings suggest the potential to develop XG014 as pan-β-CoV-B therapeutics and the importance of the XG014 conserved antigenic epitope for designing broadly protective vaccines against β-CoV-B and newly emerging SARS-CoV-2 variants of concern.
Angiotensin-Converting Enzyme 2 ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19 ; Epitopes ; Humans ; SARS-CoV-2/genetics* ; Spike Glycoprotein, Coronavirus/genetics*

TSCI have dyskinesia and sensory disturbance that can cause various life-threaten complications. The patients with traumatic spinal cord injury (TSCI), seriously affecting the quality of life of patients. Based on the epidemiology of TSCI and domestic and foreign literatures as well as expert investigations, this expert consensus reviews the definition, injury classification, rehabilitation assessment, rehabilitation strategies and rehabilitation measures of TSCI so as to provide early standardized rehabilitation treatment methods for TSCI.

Objective? To investigate the effect and safety of high-flow nasal cannula (HFNC) oxygen therapy for patients after extubation in lung transplantation. Methods? A retrospective study was conducted on 60 hospitalized patients with lung transplant during January 2017 to December 2017 in Wuxi People＇s Hospital. According to the different methods of respiratory support prescribed after extubation, the patients were divided into two groups: observation group (n=28, via HFNC) and control group (n=32, via nasal oxygen tube) . The two groups were compared in terms of the clinical indicators including their blood gas analysis(oxygenation index, lactic acid, partial pressure of carbon dioxide) , viscosity of sputum and comfort, etc. Results? Six hours after extubation and before transferring to other departments, the observation group＇s oxygenation index was(263.70±48.97)and(273.22±43.26)mmHg, which were statistically different from those of the control group with (217.83±77.30)and(229.08±68.64)mmHg respectively (P＜ 0.05). Before transferring to other departments, the partial pressure of carbon dioxide in the observation group was (37.04±8.56)mmHg, lower than the control group with (42.43±6.14)mmHg with a statistical difference (P＜0.05); the viscosity of sputum in the obseration group was lower than the control group with statistical difference (P＜0.05); the patients＇ comfort in the observation group achieved (7.72±1.06)points, higher than the control group＇s (4.39±0.82)points with statistical difference (P＜ 0.05). Conclusions? HFNC oxygen therapy for the patients after extubation in lung transplantation has good effects, is safe and reliable, and can be widely applied in clinical practice.

Objective To investigate the efficacy and safety of citrate anticoagulation for continuous renal placement therapy (CRRT) in patients at high risk of bleeding. Methods Forty-seven patients who needed to CRRT and were admitted into the department of critical care medicine from January 2015 to October 2016 were enrolled in this study. According to the patient′s actual condition, they were divided into citrate groups (24 cases) and saline group (23 cases). Patients in saline group were given saline wash. The efficacy and safety were compared between the two groups. The filter lifetime, after treatment the activated partial thromboplastin time (APTT), hemoglobin (Hb), blood gas indexes were compared between the two groups. Results The citrate group used 76 filters while the 0.9% sodium chloride group used 87 filters. The average filter lifetime in citrate group was (22.4 ± 12.7) h which was longer than (8.6±3.3) h in 0.9%sodium chloride group (t=9.79, P<0.01). The incidence of coagulation in extracorporeal circulation was 3.9%(3/76) which was lower than 16.1%(14/87) in 0.9%sodium chloride group(χ2=5.20, P<0.05). Conclusions Regional citrate anticoagulation is a safe and effective option for CRRT in patients at high risk of bleeding.

Heparinase II (Hep II) from Flavobacterium heparinum is an enzyme that could specifically cleave certain sequence of heparin and heparan sulfate. In this work, fermentation conditions of recombinant heparinase II (His-Hep II) producing bacteria were optimized, including initial induction time, inducer (IPTG) concentration, induction temperature and induction time. The optimum conditions were as follows: cultivating recombinant bacteria to exponential prophase under 37 degrees C, then adding IPTG to a final concentration of 0.3 g/L, finally cultivating recombinant bacteria under 20 degrees C for 10 h. The total crude enzyme activity reached 570 U/L. Based on these results, high cell density fermentation of recombinant bacteria was studied. The final OD600 could reach 98 and the total crude enzyme activity of His-Hep II increased to 9 436 U/L.
Fermentation ; Flavobacterium ; metabolism ; Microbiological Techniques ; Polysaccharide-Lyases ; biosynthesis ; Recombinant Proteins ; biosynthesis

Microbial nitrilases have attracted increasing attention in nitrile hydrolysis for carboxylic acid production in recent years. A bacterium with nitrilase activity was isolated and identified as Pseudomonas putida CGMCC3830 based on its morphology, physiological and biochemical characteristics, as well as 16S rRNA gene sequence. The nitrilase production was optimized by varying culture conditions using the one-factor-at-a-time method and response surface methodology. Glycerol 13.54 g/L, tryptone 11.59 g/L, yeast extract 5.21 g/L, KH2PO4 1 g/L, NaCl 1 g/L, urea 1 g/L, initial pH 6.0 and culture temperature 30 degrees C were proved to be the optimal culture conditions. It resulted in the maximal nitrilase production of 36.12 U/mL from 2.02 U/mL. Investigations on substrate specificity demonstrate P. putida nitrilase preferentially hydrolyze aromatic nitriles. When applied in nicotinic acid synthesis, 2 mg/mL P. putida cells completely hydrolyzed 20.8 g/L 3-cyanopyridine into nicotinic acid in 90 min. The results indicated P. putida CGMCC3830 displayed potential for industrial production of nicotinic acid.
Aminohydrolases ; biosynthesis ; Culture Media ; Hydrolysis ; Niacin ; biosynthesis ; Nitriles ; metabolism ; Pseudomonas putida ; enzymology ; Pyridines ; metabolism ; RNA, Ribosomal, 16S ; genetics ; Substrate Specificity ; Temperature