BACKGROUND: Previous research demonstrated that a homozygous mutation of g.136372044G>A (S12N) in caspase recruitment domain family member 9 ( CARD9 ) is critical for producing Aspergillus fumigatus -induced ( Af -induced) T helper 2 (T H 2)-mediated responses in allergic bronchopulmonary aspergillosis (ABPA). However, it remains unclear whether the CARD9S12N mutation, especially the heterozygous occurrence, predisposes the host to ABPA. METHODS: A total of 61 ABPA patients and 264 controls (including 156 healthy controls and 108 asthma patients) were recruited for sequencing the CARD9 locus to clarify whether patients with this heterozygous single-nucleotide polymorphisms are predisposed to the development of ABPA. A series of in vivo and in vitro experiments, such as quantitative real-time polymerase chain reaction, flow cytometry, and RNA isolation and quantification, were used to illuminate the involved mechanism of the disease. RESULTS: The presence of the p.S12N mutation was associated with a significant risk of ABPA in ABPA patients when compared with healthy controls and asthma patients, regardless of Aspergillus sensitivity. Relative to healthy controls without relevant allergies, the mutation of p.S12N was associated with a significant risk of ABPA (OR: 2.69 and 4.17 for GA and AA genotypes, P = 0.003 and 0.029, respectively). Compared with patients with asthma, ABPA patients had a significantly higher heterozygous mutation (GA genotype), indicating that p.S12N might be a significant ABPA-susceptibility locus ( aspergillus sensitized asthma: OR: 3.02, P = 0.009; aspergillus unsensitized asthma: OR: 2.94, P = 0.005). The mutant allele was preferentially expressed in ABPA patients with heterozygous CARD9S12N , which contributes to its functional alterations to facilitate Af -induced T H 2-mediated ABPA development. In terms of mechanism, Card9 wild-type ( Card9WT ) expression levels decreased significantly due to Af -induced decay of its messenger RNA compared to the heterozygous Card9S12N . In addition, ABPA patients with heterozygous CARD9S12N had increased Af -induced interleukin-5 production. CONCLUSION Our study provides the genetic evidence showing that the heterozygous mutation of CARD9S12N , followed by allele expression imbalance of CARD9S12N , facilitates the development of ABPA.
Humans ; Aspergillosis, Allergic Bronchopulmonary/complications* ; Aspergillus fumigatus/genetics* ; Asthma/genetics* ; Aspergillus ; Mutation/genetics* ; CARD Signaling Adaptor Proteins/genetics*

As an important element of medical and health sector innovation, the translation of scientific and technological achievements plays a key role in promoting their clinical application and meeting the medical needs of the people. The authors sorted out the problems in such translation at these affiliated hospitals in terms of " people", " finance", " material", and " system". Starting from 2017, the Tenth People′s Hospital Affiliated to Tongji University has explored such practices as establishing hospital-led clinical medical science and technology innovation parks and technology service limited companies. These practices aimed to address the issues of insufficient hospital scientific and technological innovation capabilities and the gap between the hospital′s operation mechanism to translate its scientific and technological achievements and the enterprises and the market. The clinical medical science and technology innovation park integrating administration, industry, education, research, medicine and application, has taken multiple measures to attract excellent research talents and projects from within and beyond the hospital, promote the implementation of innovative scientific research projects. The hospital also established a health industry mode with engagement of social capital from large enterprises. The Technology Services Co., Ltd. was based on the incubation and translation of hospital achievements, combining market and clinical needs, promoting multi-party cooperation between hospitals and external enterprises, improving the chain operation mechanism of hospital scientific and technological achievements translation work, and alleviating the problem of insufficient research pilot funds and productibility funds by means of hospital-led fundraising. The number of patent authorizations of hospitals had increased from 23 cases in 2018 to 105 in 2022, and the amount of patent conversion had increased from 2 million yuan in 2020 to 11 million yuan in 2022. It is recommended that affiliated hospitals of universities further improve the organizational structure of achievement translation, strengthen their professional talent teams, improve their operation mechanism of achievement translation, build a platform for medical school-enterprise cooperation, and improve the evaluation mechanism of translation assessment, in order to promote a virtuous cycle of hospital′s scientific and technological achievement translation work.

Aristolochic acids (AAs) have long been considered as a potent carcinogen due to its nephrotoxicity. Aristolochic acid I (AAI) reacts with DNA to form covalent aristolactam (AL)-DNA adducts, leading to subsequent A to T transversion mutation, commonly referred as AA mutational signature. Previous research inferred that AAs were widely implicated in liver cancer throughout Asia. In this study, we explored whether AAs exposure was the main cause of liver cancer in the context of HBV infection in mainland China. Totally 1256 liver cancer samples were randomly retrieved from 3 medical centers and a refined bioanalytical method was used to detect AAI-DNA adducts. 5.10% of these samples could be identified as AAI positive exposure. Whole genome sequencing suggested 8.41% of 107 liver cancer patients exhibited the dominant AA mutational signature, indicating a relatively low overall AAI exposure rate. In animal models, long-term administration of AAI barely increased liver tumorigenesis in adult mice, opposite from its tumor-inducing role when subjected to infant mice. Furthermore, AAI induced dose-dependent accumulation of AA-DNA adduct in target organs in adult mice, with the most detected in kidney instead of liver. Taken together, our data indicate that AA exposure was not the major threat of liver cancer in adulthood.

Glutamine synthetase is a key enzyme in plant nitrogen assimilation. To study the structure of wheat glutamine synthetase isoenzymes, GS1, GSr, GSe, GS2 and GS2p of wheat were cloned into pET-21a, and the expression condition was optimized. Although wheat glutamine synthetase isoenzymes had 70%-80% amino acid sequence homology, the isoforms expressed with different characteristics. Induced at 30 °C, the most expression level of GSr, GSe and GS2 was after 3 h, and of GS1 was at the 7 h whereas no GS2p was expressed, and the GS isoenzymes showed different expression level, with the order of GS1 (22%)>GSr (15%)>GS2 (12%)>GSe (5%). GSe expressed as soluble protein, and GS1 expressed mainly as soluble protein whereas GSr and GS2 expressed as insoluble proteins. Induced at 30 °C for 3 h, mRNA transcript levels of GS isoforms were different, with the order of GSr (7.59)>GS2 (1.84)>GS2p (1.66)>GSe (1.46)>GS1 (1.00). The levels of mRNA transcription were not consistent with the level of the protein translation. The analysis of mRNA secondary structure showed the free energy of translation initiation region of glutamine synthetase isoforms was different, with the order of GS1 (14.4)GSr (13%)>GS2 (10%)>GSe (7%), and different activities with GS1>GSe>GS2, and the activity of GSr was not detected. The gene sequence of glutamine synthetase isoenzymes determines the amount, status and activity of proteins expressed in prokaryotic cells.

Keratinized mucosa in oral cavity plays an important role in periodontal health. The defect of keratinized mucosa may increase the risks of complication of oral implant surgery and restoration. Graft of keratinized tissue and connective tissue are still the gold standard for treating keratinized mucosa defect now. The current research focus on how to modulate non-keratinized mucosa to highly-efficient and minimally-invasive keratinized mucosa. Keratinocytes are critical components of oral mucosa and its final differentiation into keratinized mucosa is controlled by the connective tissue microenvironment involving a variety of molecules and ions. To fully understand keratinized differentiation of keratinocyte, this review focuses on its influence factors and possible mechanisms under the differentiation.

Objective To evaluate the in vitro effect on tumor cell uptake,tumor imaging and in vivo biodistribution of 99Tcm-epidermal growth factor receptor (EGFR) mRNA antisense PNA probe mediated by cationic liposome.Methods The oligonucleotide with sequence complementary to part of the EGFR mRNA antisense PNA was hybridized in an anti-parallel orientation targeted PNA.PNA hybridization complexes were labeled with 99Tcm by ligand exchange.The assembly of lipofectamine and 99Tcm-labeled heteroduplex was achieved by electrostatic interactions,and the radiolabeled purity was determined by reversedphase HPLC (RP-HPLC).The disparities of cell uptake in SKOV3 cells and the differences of biodistribution and molecular imaging in BALB/c nude mice bearing SKOV3 xenografts between lipofectanine-mediated 99Tcm-EGFR mRNA antisense PNA (group 1) and 99Tcm-EGFR mRNA antisense PNA (group 2) were analyzed.Two-sample t (or t') test and Wilcoxon rank sum test were used for statistical analysis.Results The labeling rates of both group 1 and group 2 were more than 95％ within 6 h.The cell uptake at 1,2,4,6,12,24 h after injection was (28.90±1.12)％,(32.76±1.20)％,(38.20±3.11)％,(41.23±1.60)％,(46.63±1.55)％ and (46.78±2.14)％ in group 1,and was (3.51±0.39)％,(3.90±0.40)％,(4.69±0.18)％,(5.91±0.26)％,(5.30±0.22)％ and (5.39±0.17)％ in group 2 respectively (t'=47.11-58.67,Z=2.80,all P＜0.05).The retention ratios showed significant difference between the two groups (t'=7.25-11.55,Z=2.80,all P＜0.05).The SKOV3 tumor could be visualized in both groups at 1 h post injection but much better visualized in group 1.The T/NT ratios were higher in group 1 at all time points (t =3.96,t'=12.65-14.69,Z=2.83-5.29,all P＜0.05).The T/NT ratios at uptake peak were 5.02 and 3.95,respectively.The probe accumulated mainly in tumor,kidneys and liver.Tumor uptake increased with time ((1.49±0.09) ％ID/g and (2.15±0.21) ％ID/g at 1 h,(3.90±0.65) ％ID/g and (5.00±0.10) ％ID/g at 6 h) after lipofectamine treatment.The ratios of tumor to contralateral muscle were also higher in group 1 (t =11.24,t' =3.96-11.94,all P＜0.05).Conclusions Lipofectamine-mediation can significantly improve the intracellular delivery of radionuclide molecular probe.Lipofectamine-mediated 99Tcm-EGFR mRNA antisense PNA can greatly improve imaging contrast and visualization of EGFR-over-expressing tumors.

Objective To prepare the 99Tcm-labeled human epidermal growth factor receptor type 2 (HER2) affibody molecule ZHER2:342 and evaluate its receptor binding specificity in vitro.Methods The molecular ZHERa:342 was labeled with 99Tcm using the ligand exchange method.The labeling efficiency and radiochemical purity were measured by HPLC.The major factors,such as the mass of SnC12 and NaOH and reaction time were analyzed,and the optimal method was summarized.Cell binding kinetics and cellular retention of the probe were investigated in HER2-expressing SKOV-3 cells and MDA-MB-231 cells with low HER2 expression respectively.HER2 binding specificity of 99Tcm-ZHER2:342 was analyzed by a pre-injection of excess unlabeled ZHER2:342 to saturate HER2 receptors.One-way analysis of variance and two-sample t test were used.Results The optimal labeling procedure was as follows:5 μg (1 g/L) of ZHER2:342 was mixed with 5 μg of NaOH (1 g/L),then 8.8 μg SnC12(1 g/L,solution) was added,followed by 150 μl (37 MBq) 99TcmO4-solution,and finally the mixture was slightly vortexed and incubated for 1 h at room temperature.99TcmZHER2:342 was stable in vitro with a high labeling efficiency of (98.10± 1.73)％.The radiochemical purity was ＞ 98％,and was more than 85％ after the incubation for 24 h in saline and fresh human serum.The cell binding of 99Tcm-ZHER2:342 with HER2-expressing SKOV-3 cells gradually increased over time with a peak of (9.95± 1.02)％ at 6 h.The binding of 99Tcm-ZHER2:342 in SKOV-3 cells was significantly higher than that in MDA-MB-231 cells at every time point (5.68-9.88 vs 0.56-2.11 ; t:from-34.50 to-13.14,all P＜0.01).The labeled molecular probe retained the capacity to bind specifically to HER2-expressing SKOV-3 cells since the cell binding decreased from (9.95 ± 1.02) ％ to (2.11 ±0.27) ％ after receptor saturation (t =-13.14,P＜0.01).Conclusions 99Tcm-ZHER2:342 has a high labeling efficiency,good stability and optimal binding specificity.These characteristics enable it to be a promising molecular probe for HER2-targeting imaging.

A total of 38 cases of senile brucella spondylitis disease at our hospital during January 2002 and March 2012 were analyzed .After admission , all of them were definitely diagnosed on the basis of epidemiological history , clinical manifestations , laboratory tests , imaging and pathological examinations . Over a follow-up period of 12 months, 17 cases were cured after standardized drug treatment .Among 21 surgical cases, there were curing (n=17) and improving (n=2).Senile brucellosis spondylitis has distinct serological and pathological characteristics .And formulating the diagnostic criteria may improve its diagnostic rate and reduce its misdiagnostic rate .And standardized drug therapy achieves a better curing rate and a proper timing of surgical intervention improves its clinical outcomes .

The purpose of this paper was to evaluate the relationship between the thrombosis and secretory duct dilation,lesion size,clinical types,nature (primary or recurrent) and duration of illness in the development of ranula.A total of 229 cases of sublingual gland cysts were treated with surgical resection from Jan.1990 to Feb.2010.The patients' data were investigated on histopathological findings,size of ranula,the clinical types,nature of ranula (primary or recurrent) and duration of illness.Sections from the paraffin-embeded blocks were HE-stained.CK expression was immunohistochemically detected.Among 229 cases the incidence of venous thrombosis was 58.52％.The incidence of venous thrombosis with or without duct dilation was 73.25％ and 26.39％ respectively,with a significant difference between the two groups (P＜0.005).The incidence of venous thrombosis of ranulas with diameter larger or less than 3 cm was 72.22％ and 46.28％ (P＜0.005).The incidence of venous thrombosis of oral ranula,plunging ranula and mixed ranula was 49.37％,77.19％ and 85.71％ respectively,with a significant difference found between oral and plunging or mixed ranula (P＜0.01).The incidence of venous thrombosis in ranula patients with duration of illness longer or less than 3 months was 69.77％ and 51.75％ (P＜0.01).The incidence of venous thrombosis with recurrent and primary ranulas was 51.85％ and 64.85％,without a significant difference noted between them (P＞0.05).It is concluded that the formation of venous thrombosis was related to the dilation of secretory duct,lesion size,clinical types,duration of lesion but formation of venous thrombosis was not related to the nature (primary or recurrent) of ranulas.

The purpose of this paper was to evaluate the relationship between the thrombosis and secretory duct dilation, lesion size, clinical types, nature (primary or recurrent) and duration of illness in the development of ranula. A total of 229 cases of sublingual gland cysts were treated with surgical resection from Jan. 1990 to Feb. 2010. The patients' data were investigated on histopathological findings, size of ranula, the clinical types, nature of ranula (primary or recurrent) and duration of illness. Sections from the paraffin-embeded blocks were HE-stained. CK expression was immunohistochemically detected. Among 229 cases the incidence of venous thrombosis was 58.52%. The incidence of venous thrombosis with or without duct dilation was 73.25% and 26.39% respectively, with a significant difference between the two groups (P<0.005). The incidence of venous thrombosis of ranulas with diameter larger or less than 3 cm was 72.22% and 46.28% (P<0.005). The incidence of venous thrombosis of oral ranula, plunging ranula and mixed ranula was 49.37%, 77.19% and 85.71% respectively, with a significant difference found between oral and plunging or mixed ranula (P<0.01). The incidence of venous thrombosis in ranula patients with duration of illness longer or less than 3 months was 69.77% and 51.75% (P<0.01). The incidence of venous thrombosis with recurrent and primary ranulas was 51.85% and 64.85%, without a significant difference noted between them (P>0.05). It is concluded that the formation of venous thrombosis was related to the dilation of secretory duct, lesion size, clinical types, duration of lesion but formation of venous thrombosis was not related to the nature (primary or recurrent) of ranulas.