Objective To identify the pathogenic gene and variant for a family with branchio-otic syndrome.Methods The clinical data of this family were collected,and the peripheral blood was extracted for deafness gene NGS panel analysis.Pathogenic variation detected was verified by Sanger sequencing.Results The family contained 17 members in three-generations,3 of whom exhibited autosomal dominant,hearing loss,preauricular fistula and branchial cleft fistula,which were in accordance with the clinical diagnosis criteria of branchio-otic syndrome.A no-vel heterozygous variant c.963dupT(p.E322X)in EYA1 gene was identified,which co-segregated with the branchi-o-otic syndrome phenotype in the family.The variant was a nonsense variant resulting in the premature appearance of the stop codon.According to the American College of Medical Genetics and Genomics(ACMG)guidelines and the criteria,the variant was classified as pathogenic.Conclusion We identified a novel pathogenic variant EYA1:c.963dupT(p.E322X)in a family with branchio-otic syndrome.

Objective To identify the pathogenic gene and variant for a family with branchio-otic syndrome.Methods The clinical data of this family were collected,and the peripheral blood was extracted for deafness gene NGS panel analysis.Pathogenic variation detected was verified by Sanger sequencing.Results The family contained 17 members in three-generations,3 of whom exhibited autosomal dominant,hearing loss,preauricular fistula and branchial cleft fistula,which were in accordance with the clinical diagnosis criteria of branchio-otic syndrome.A no-vel heterozygous variant c.963dupT(p.E322X)in EYA1 gene was identified,which co-segregated with the branchi-o-otic syndrome phenotype in the family.The variant was a nonsense variant resulting in the premature appearance of the stop codon.According to the American College of Medical Genetics and Genomics(ACMG)guidelines and the criteria,the variant was classified as pathogenic.Conclusion We identified a novel pathogenic variant EYA1:c.963dupT(p.E322X)in a family with branchio-otic syndrome.

Poor water solubility is the primary obstacle preventing the development of many pharmacologically active compounds into oral preparations. Self-nanoemulsifying drug delivery systems(SNEDDS) have become a widely used strategy to enhance the oral bioavailability of poorly soluble drugs by inducing a supersaturated state, thereby improving their apparent solubility and dissolution rate. However, the supersaturated solutions formed in SNEDDS are thermodynamically unstable systems with solubility levels exceeding the crystalline equilibrium solubility, making them prone to drug precipitation in the gastrointestinal tract and ultimately hindering drug absorption. Therefore, maintaining a stable supersaturated state is crucial for the effective delivery of poorly soluble drugs. Incorporating polymers as precipitation inhibitors(PPIs) into the formulation of supersaturated self-nanoemulsifying drug delivery systems(S-SNEDDS) can inhibit drug aggregation and crystallization, thus maintaining a stable supersaturated state. This has emerged as a novel preparation strategy and a key focus in SNEDDS research. This review explores the preparation design of SNEDDS and the technical challenges involved, with a particular focus on polymer-based S-SNEDDS for enhancing the solubility and oral bioavailability of poorly soluble drugs. It further elucidates the mechanisms by which polymers participate in transmembrane transport, summarizes the principles by which polymers sustain a supersaturated state, and discusses strategies for enhancing drug absorption. Altogether, this review provides a structured framework for the development of S-SNEDDS preparations with stable quality and reduced development risk, and offers a theoretical reference for the application of S-SNEDDS technology in improving the oral bioavailability of poorly soluble drugs.
Solubility ; Administration, Oral ; Polymers/chemistry* ; Drug Delivery Systems/methods* ; Humans ; Emulsions/chemistry* ; Biological Availability ; Animals ; Pharmaceutical Preparations/administration & dosage*

Objective:To study the methylation level of miR-5194 promoter in bladder cancer tissues, and explore the effects of miR-5194 on the proliferation and migration of bladder cancer cells by targeting p21-activated protein kinase 2 (PAK2).Methods:The methylation level of miR-5194 promoter in bladder cancer tissues was analyzed using MethHC database. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-5194 in bladder cancer MGH-U3, EJ, J82, and UMUC3 cells. 5-aza-2′-deoxycytidine (5-Aza-CdR) was used to treat bladder cancer cell lines, and RT-qPCR was used to detect the changes in the expression of miR-5194 in bladder cancer cell lines after 5-Aza-CdR treatment. UMUC3 cells were divided into miR-5194 group and NC group, and miR-5194 or miR-NC were transfected into UMUC3 cells, respectively. Colony formation assay and scratch assay were used to detect the effect of overexpression of miR-5194 on the proliferation and migration of UMUC3 cells. The bioinformatics tool miRGator and dual-luciferase reporter gene experiments verified the targeting relationship between miR-5194 and PAK2. The effect of overexpression of miR-5194 on the expression of PAK2 mRNA in UMUC3 cells was detected by RT-qPCR. The effect of overexpression of miR-5194 on the expression of PAK2 protein, proliferation-related proteins (CDK1, Cyclin B) and migration-related proteins (FOXC2, E47) in UMUC3 cells was detected by Western blotting. The measurement data were expressed as mean ± standard deviation ( ± s), the independent sample t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison among multiple groups. Results:The methylation level of miR-5194 promoter in bladder cancer tissues was significantly higher than that in adjacent tissues ( P<0.01). Compared with the immortalized bladder epithelial cells SV-HUC-1, the expression of miR-5194 in bladder cancer cells was significantly down-regulated ( P<0.01). After 5-Aza-CdR treatment, the expression of miR-5194 in bladder cancer cells was significantly increased ( P<0.01). The number of colonies in miR-5194 group and NC group were 31.30 ± 8.09 and 99.98 ± 10.53, respectively, and the proliferation ability of UMUC3 cells in miR-5194 group was weakened ( P<0.01). The migration rates of UMUC3 cells in miR-5194 group and NC group were (31.50 ± 7.17)% and (76.06 ± 4.86)%, respectively, and the migration ability of UMUC3 cells in miR-5194 group was weakened ( P<0.01). miR-5194 can target bind PAK2 gene ( P<0.01). The relative expression of PAK2 mRNA in UMUC3 cells of miR-5194 group and NC group were 1.02 ± 0.34 and 5.43 ± 0.76, respectively, and miR-5194 could negatively regulate the expression of PAK2 mRNA ( P<0.01). Compared with the NC group, the expression of PAK2 protein, the expression of proliferation-related proteins CDK1 and Cyclin B, and the expression of migration-related proteins FOXC2 and E47 were down-regulated in UMUC3 cells with miR-5194 overexpression. Conclusion:The methylation level of miR-5194 promoter in bladder cancer tissue was significantly increased, and miR-5194 inhibited the proliferation and migration of bladder cancer cells by targeting down-regulation of PAK2 expression in bladder cancer UMUC3 cells.

Objective:To investigate the risk factors of amputation in patients with diabetic foot ulcer (DFU) in order to improve the prognosis and reduce the amputation rate.Methods:The clinical data of 359 DFU patients admitted to the People′s Hospital of Xinjiang Uygur Autonomous Region from January 2017 to August 2021 were retrospectively analyzed, and they were divided into amputation group (161 cases) and non-amputation group (198 cases) according to whether amputation surgery was performed. Demographic characteristics, Wagner grading and other data of the two groups were collected. Forward step logistic regression analysis was used to identify independent risk factors for amputation, and receiver operating characteristic (ROC) curves were used to evaluate the predictive value of each risk factor for amputation in DFU patients.Results:There were significant differences between the amputation and non-amputation groups in terms of previous amputation history, peripheral vascular diseases (PVD), diabetic foot secondary osteomyelitis, diabetic nephropathy (DN), history of angioplasty, Wanger grade, K +, age, white blood cell count, C-reactive protein, high density lipoprotein cholesterol (HDL-C), estimated glomerular filtration rate, cardiac troponin T, and cardiac troponin I, lactic acid (all P<0.05). Previous amputation history ( OR=2.329, 95% CI: 1.092-4.970, P=0.029), DN ( OR=4.091, 95% CI: 2.222-7.532, P<0.001), PVD ( OR=2.556, 95% CI: 1.487-4.395, P=0.001), diabetic foot secondary osteomyelitis ( OR=6.332, 95% CI: 3.595-11.153, P<0.001), Wagner grade were independent risk factors for amputation in DFU patients, HDL-C ( OR=0.392, 95% CI: 0.182-0.842, P=0.016) were protective factors for amputation in DFU patients. Moreover, the combined accuracy of the above factors in predicting amputation in DFU patients was high, and the area under ROC curve was 0.839 (95% CI: 0.798-0.880), sensitivity was 83%, and specificity was 73% ( OR=0.05). Conclusions:Previous amputation history, DN, PVD, diabetic foot secondary osteomyelitis and Wagner grade are independent risk factors for amputation in DFU patients, while HDL-C is a protective factor for amputation in DFU patients. Further investigation will help to establish a stratified system for predicting the risk of diabetic foot, so as to achieve better individualized treatment.

Objective:To investigate the risk factors of amputation in patients with diabetic foot ulcer (DFU) in order to improve the prognosis and reduce the amputation rate.Methods:The clinical data of 359 DFU patients admitted to the People′s Hospital of Xinjiang Uygur Autonomous Region from January 2017 to August 2021 were retrospectively analyzed, and they were divided into amputation group (161 cases) and non-amputation group (198 cases) according to whether amputation surgery was performed. Demographic characteristics, Wagner grading and other data of the two groups were collected. Forward step logistic regression analysis was used to identify independent risk factors for amputation, and receiver operating characteristic (ROC) curves were used to evaluate the predictive value of each risk factor for amputation in DFU patients.Results:There were significant differences between the amputation and non-amputation groups in terms of previous amputation history, peripheral vascular diseases (PVD), diabetic foot secondary osteomyelitis, diabetic nephropathy (DN), history of angioplasty, Wanger grade, K +, age, white blood cell count, C-reactive protein, high density lipoprotein cholesterol (HDL-C), estimated glomerular filtration rate, cardiac troponin T, and cardiac troponin I, lactic acid (all P<0.05). Previous amputation history ( OR=2.329, 95% CI: 1.092-4.970, P=0.029), DN ( OR=4.091, 95% CI: 2.222-7.532, P<0.001), PVD ( OR=2.556, 95% CI: 1.487-4.395, P=0.001), diabetic foot secondary osteomyelitis ( OR=6.332, 95% CI: 3.595-11.153, P<0.001), Wagner grade were independent risk factors for amputation in DFU patients, HDL-C ( OR=0.392, 95% CI: 0.182-0.842, P=0.016) were protective factors for amputation in DFU patients. Moreover, the combined accuracy of the above factors in predicting amputation in DFU patients was high, and the area under ROC curve was 0.839 (95% CI: 0.798-0.880), sensitivity was 83%, and specificity was 73% ( OR=0.05). Conclusions:Previous amputation history, DN, PVD, diabetic foot secondary osteomyelitis and Wagner grade are independent risk factors for amputation in DFU patients, while HDL-C is a protective factor for amputation in DFU patients. Further investigation will help to establish a stratified system for predicting the risk of diabetic foot, so as to achieve better individualized treatment.

Objective To observe the clinical efficacy of low-dose dobutamine combined with tolvaptan in the treatment of type Ⅰcardiorenal syndrome.Methods A total of 104 patients with type 1 cardiorenal syndrome admitted to the Department of Cardiology,Xinhua Hospital Affiliated to Shanghai Jiao Tong University of Medicine were selected as study subjects.The patients were randomly divid-ed into control group and experimental group,52 patients in each group.The control group which accepted oral tolvaptan 15mg,and the experimental group which accepted the treatment of low-dose of dopamine[2μg/(kg·min)]with intravenous infusion,and they were treatment for 7d.The primary endpoint was the cardiorenal serological index after 7days of treatment.Results After treatment,the total effective rates of experimental group and control group were 86.53％(45/52)and 67.30％(35/52),respectively,and the difference was statistically significant(P＜0.05).After treatment,the serological levels of cardiorenal functions and 12h urine volume were more significantly improved in two groups,and the improvement effect of experimental group was better than that of control group,the difference was statistically significant(P＜0.05).The levels of serum inflammatory cytokines after treatment were significantly lower than before treatment,and the difference was statistically significant(P＜0.05).Conclusion Low-dose dopamine combined with tolvaptan in treating type 1 cardio-renal syndrome can significantly improve patients'cardiac and renal function.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To investigate whether ultra-high dose rate (FLASH) irradiation can reduce radiation-induced intestinal injuries of mice compared to conventional dose rate (CONV) irradiation.Methods:Both FLASH and CONV irradiation were delivered with electron beam, with dose rates of 750 Gy/s and 0.5G y/s, respectively. A total of 105 mice were randomly divided into groups using a simple randomization method. Twenty-one mice were selected for weight observation, 7 mice in each group. After 9 Gy FLASH and CONV irradiation on the abdomen, the weight changes of mice were measured every other day, and compared among three groups. Twenty-four mice were selected for pathological examination including 5 mice in the control group. Three-and-a-half-day days after 12 Gy FLASH ( n=10) and CONV irradiation ( n=9) on the abdomen, the intestines of the mice were taken. Pathological sections were stained with hematoxylin-eosin (HE) to compare the number and percentage of regenerated crypts of the small intestine between two groups. After 12 Gy FLASH ( n=10) and CONV irradiation ( n=10) on the abdomen, the survival of 20 mice was observed. After FLASH using 4.5 Gy×2 times ( n=10) and CONV irradiation at 9 Gy×1 time ( n=10) on the abdomen, the weight changes were observed. After FLASH using 6 Gy×2 times ( n=10) and CONV irradiation at 12 Gy×1 time ( n=10) on the abdomen, the survival of mice was observed. The time interval between two irradiation was 1 min. EBT3 film was employed to monitor the actual exposure dose of the mice. The variables conforming to normal distribution were expressed by Mean±SD. Inter group comparison was performed by independent t-test. The survival of mice among different groups was compared by log-rank test. Results:After 9 Gy of abdominal irradiation, the mean weight of mice in the FLASH group was significantly higher than that in the CONV group. The weight of mice in the FLASH and CONV groups was (19.8±0.8) g and (18.0±1.8)g ( P=0.036) at 7 days after irradiation, (22.0±1.0)g and (21.2±0.5)g ( P=0.075) at 15 days after irradiation, and (24.2±1.4)g and (22.0±1.2)g ( P=0.012) at 25 days after irradiation, respectively. After 12 Gy irradiation, the mean survival of mice in FLASH and CONV groups was 4 days and 4.7 days ( P=0.029). After 12 Gy total abdominal irradiation, the mean number of intestinal regenerative crypts in the FLASH and CONV groups was 2.9/mm and 1.2/mm ( P=0.041), and the percentage of intestinal regenerative crypts was 34.1% and 14.1%, respectively. The survival of mice irradiated by FLASH using 6 Gy×2 times was longer compared with that of mice after CONV irradiation at 12 Gy×1 time. The weight of mice after 4.5 Gy×2 times irradiation was higher than that of mice after CONV irradiation at 9 Gy×1 time. Conclusion:Weight, survival and the number of intestinal regenerative crypts in the FLASH group are higher than those in the CONV group after irradiation, indicating that radiation-induced intestinal injury caused by FLASH irradiation is slighter than that of CONV irradiation.

The last essential enzyme in the biosynthetic pathway of trilobatin, phloretin-4'-O glycosyltransferase (P4'-OGT), catalyzes the conversion of trilobatin to phloretin in vitro. However, only a few P4'-OGTs have been found in plants. This study used Malus domestica phloretin-4'-O glycosyltransferase (MdPh-4'-OGT) as a query to identify and clone two UDP-glucuronosyltransferase (UGT) genes, designated UGT74L2 and UGT74L3, from the transcriptome of Andrographis paniculata. According to a phylogenetic tree analysis, UGT74L2 and UGT74L3 belonged to the UGT74 family, which has been linked to several activities in other species. The in vitro enzymatic reaction demonstrated that UGT74L2 could particularly catalyze the formation of trilobatin from phloretin, but UGT74L3 had no effects. By using Ni-NTA affinity chromatography to extract the soluble UGT74L2 recombinant protein, the enzymatic kinetics of the activity was investigated using phloretin as the substrate. The results showed that the optimal temperature and pH for UGT74L2 enzymatic reaction were 40 ℃ and 8.0 (Tris-HCl system), respectively. Three metal ions (Ca²⁺, Mn²⁺ and Co²⁺) showed inhibitory effect on the activity of UGT74L2, while Mg²⁺ could improve the activity of UGT74L2. Other tested metal ions have no significant effect on UGT74L2. The results of enzymatic kinetic parameters that the K_m value was 29.84 μmol·L^-1, the k_cat was 0.02 s^-1, and the k_cat·K_m^-1 was 572.6 mol^-1·s^-1. By homology modeling, molecular docking and mutation experiments, we found that multiple amino acids residues around the substrate binding pocket play quite an important role during catalytic process, In summary, we identified a novel P4'-OGT gene from medicinal plant Andrographis paniculata and provided a new efficient catalyst to synthesize trilobatin. Meanwhile, this study provides a reference for mining new efficient glycosylation modules from plants.