Objective To investigate the mechanism by which Xinqian Ganjie Decoction allevi-ates inflammatory responses in rat model of chronic rhinosinusitis by inhibiting the activation of the toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signaling pathway.Methods Thirty-two 4-week-old healthy male Sprague-Dawley(SD)rats were randomly divided into model group(intragas-tric administration of 1.5 mL normal saline)and low-,medium-,and high-dose groups(intragastric administration of 0.75,1.50,and 3.00 mL/kg Xinqian Ganjie Decoction,respectively),with 8 rats in each group.Another 8 healthy rats were randomly selected as control group(intragastric adminis-tration of 1.5 mL normal saline).A rat model of chronic rhinosinusitis was established in the model group and the low-,medium-,and high-dose groups by placing expanded cotton pledgets in the nasal cavity and injecting Staphylococcus aureus suspension.After successful model establishment,rats in each group were treated by intragastric administration for 14 consecutive days.The number of snee-zes and nose scratches in each group was observed.Olfactory function changes in rats were assessed using an olfactory function test.Histopathological changes in nasal mucosa were detected by hema-toxylin-eosin(HE)staining.The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in nasal mucosa were measured by enzyme-linked immunosor-bent assay(ELISA).The expression levels of proteins related to the TLR4/NF-κB signaling path-way were detected by western blot.Results Compared with the control group,rats in the model,low-,medium-,and high-dose groups showed increased number of sneezes and nose scratches.Compared with the model group,rats in the medium-and high-dose groups exhibited decreased number of sneezes and nose scratches.The differences among these groups were all statistically sig-nificant(P＜0.01).Before treatment,the time taken by rats to find food pellets in the control group was longer than that in the other groups(P＜0.01).After treatment,rats took longer to find food pellets in the model group compared with the control group,while rats in the low-,medium-,and high-dose groups took less time than the model group,with a decreasing trend in time as the dose increased.The differences among these groups were all statistically significant(P＜0.01).HE staining results showed that the nasal mucosa of rats in the control group was intact with no in-flammatory cell infiltration.Compared with the control group,the nasal mucosa of rats in the model group was significantly thickened with a large number of inflammatory cell infiltrations.Compared with the model group,the nasal mucosa thickening in rats of the medium-and high-dose groups was alleviated,with only a small amount of inflammatory cell infiltration observed.The levels of TNF-α,IL-1β,and IL-6 in the nasal mucosa of rats in the model group were higher than those in the control group.Compared with the model group,the levels of TNF-α,IL-1β,and IL-6 in the medium-and high-dose groups were decreased.The above differences among these groups were all statistically sig-nificant(P＜0.01).The expression levels of TLR4 and p-p65 NF-κB proteins in the nasal mucosa of rats in the model group were higher than those in the control group.Compared with the model group,the expression levels of TLR4 and p-p65 NF-κB in the medium-and high-dose groups were decreased.The differences among these groups were all statistically significant(P＜0.01).Con-clusion Xinqian Ganjie Decoction may reduce the release of TNF-α,IL-1β,and IL-6 by inhibiting the activation of the TLR4/NF-κB signaling pathway,thereby alleviating inflammatory responses and improving symptoms in a rat model of CRS.

Objective:To evaluate the value of ultrasonography in the differential diagnosis of cystic biliary atresia (CBA) and choledochal cyst (CC) in infants.Methods:CBA or CC children <3 months of age diagnosed by surgery in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from 2015 to 2023 were collected, and the differences in general conditions and ultrasound manifestations between CBA group and CC group were retrospectively analyzed.Results:Among the 55 children, 30 were in CBA group and 25 were in CC group. The mean length diameter and width diameter of cysts in the CBA group were about (1.45±1.05)cm and (1.04±0.73)cm, respectively, which were significantly smaller than those in the CC group (5.41±2.98)cm and (3.21±2.90)cm ( P<0.001). The incidence of fibrous plaque (50.0%, 15/30) and abnormal gallbladder morphology (73.3%, 22/30) in the CBA group was significantly higher than that in the CC group [4.0%(1/25) and 16.0%(4/25), respectively]. The incidence of intrahepatic biliary mud deposition (52.0%, 13/25) and intrahepatic biliary duct dilation (64.0%, 16/25) in the CC group was significantly higher than that in the CBA group [0, 3.3%(1/30), respectively]. Conclusions:There are some differences in ultrasonography between CBA and CC children. The length and width of cysts in children with CC are significantly larger than those in children with CBA, and biliary mud deposits or intrahepatic bile duct dilatation are seen in the cysts. The incidence of fibrous plaque and abnormal gallbladder morphology in CBA children is significantly higher than that in CC children.

Objective To quantify the setup errors for the different anatomical sites of patients who received intensity-modulated radiotherapy (IMRT) with linear accelerator on-board kilovolt fan beam CT(kV-FBCT) as non-isocenter IGRT and megavolt cone beam CT (MV-CBCT) as isocenter IGRT. Methods A retrospective analysis was performedon 70 patients who underwent radiotherapy, kV-FBCT, and/or MV-CBCT scans after each routine setup prior to IMRT. The average displacement (M), systematic error (Σ), and random error (б) at different treatment sites in the left-right, anterior-posterior, and cranial-caudal directions were calculated according to the individual displacements. The formula 2.5Σ+0.7б was used to estimate the PTV margin in respective direction. For each single patient, the root mean square in three directions was used as 3D displacement. Results A total of 1130 displacements were recorded in the 70 patients. The PTV margin was estimated to be 1.9-3.1 mm in head and neck cancer, 2.8-5.1 mm in thoracic cancer, 4.6-5.1 mm in breast cancer, 3.0-5.5 mm in upper abdominal cancer, and 3.5-6.8 mm in pelvic tumor. For the 3D mean displacements, the head and neck, thoracic, breast, upper abdominal, and pelvic cancer were 2.4±1.0, 4.0±1.6, 4.1±2.0, 4.6±2.1, and 4.6±2.1 mm, respectively. The average 3D displacement obtained by kV-FBCT and MV-CBCT were 4.1 and 3.4 mm, respectively (P=0.212). Conclusion The quantitative setup-error data can be obtained using linear accelerator on-board FBCT, and the non-isocenter IGRT induced set-up error cannot be negligible.

As a ligand-dependent transcription factor,retinoid-associated orphan receptor γt(RORyt)that controls T helper(Th)17 cell differentiation and interleukin(IL)-17 expression plays a critical role in the pro-gression of several inflammatory and autoimmune conditions.An emerging novel approach to the therapy of these diseases thus involves controlling the transcriptional capacity of RORyt to decrease Th17 cell development and IL-17 production.Several RORyt inhibitors including both antagonists and inverse agonists have been discovered to regulate the transcriptional activity of RORyt by binding to orthosteric-or allosteric-binding sites in the ligand-binding domain.Some of small-molecule inhibitors have entered clinical evaluations.Therefore,in current review,the role of RORyt in Th17 regulation and Th17-related inflammatory and autoimmune diseases was highlighted.Notably,the recently developed RORyt inhibitors were summarized,with an emphasis on their optimization from lead compounds,ef-ficacy,toxicity,mechanisms of action,and clinical trials.The limitations of current development in this area were also discussed to facilitate future research.

Ulcerative colitis (UC) is one of types of inflammatory bowel disease with high recurrence. Recent studies have highlighted that microbial dysbiosis as well as abnormal gut immunity are crucial factors that initiate a series of inflammatory responses in the UC. Modulating the gut microbiota-intestinal immunity loop has been suggested as one of key strategies for relieving UC. Many Chinese herbal medicines including some of single herb, herbal formulas and the derived constituents have been reported with protective effect against UC through modulating gut microbiome and intestinal immunity. Some clinical trials have shown promising results. This review thus focused on the current knowledge on using Chinese herbal medicines for treating UC from the mechanism aspects of regulating intestinal homeostasis involving microbiota and gut immunity. The existing clinical trials are also summarized.

Enzyme-catalysis self-assembled oligopeptide hydrogel holds great interest in drug delivery, which has merits of biocompatibility, biodegradability and mild gelation conditions. However, its application for protein delivery is greatly limited by inevitable degradation of enzyme on the encapsulated proteins leading to loss of protein activity. Moreover, for the intracellularly acted proteins, cell membrane as a primary barrier hinders the transmembrane delivery of proteins. The internalized proteins also suffer from acidic and enzymatic degradation in endosomes and lysosomes. We herein develop a protease-manipulated hybrid nanogel/nanofiber hydrogel for localized delivery of intracellularly acted proteins. The embedded polymeric nanogels (CytoC/aNGs) preserve activity of cytochrome

Objective:To evaluate the feasibility and clinical efficacy of deformed complex vertebral osteotomy (DCVO) technique on the treatment of angular kyphosis of cured spinal tuberculosis.Methods:A retrospective study was performed on patients with angular kyphosis of cured spinal tuberculosis who underwent the DCVO technique or posterior vertebral column resection (PVCR) technique from Jan, 2007 to Jan, 2019. 33 patients were included, 18 males and 15 females, the average age was 39.5±15.0 years old (ranged 9-78 years old). The vertebral deformity in thoracic vertebrae 14 cases, thoracolumbar vertebrae 16 cases, and lumbar vertebrae 3 cases. 20 cases underwent the DCVO technique, while 13 cases underwent PVCR technique. For DCVO group, the multiple malformed vertebrae were considered a malformed complex, and a larger range and angle wedge osteotomy was performed within the complex using the DCVO technique. PVCR technique would resect the whole deformed vertebrae, and subsequently brought the two separated spinal columns together with instruments and titanium mesh. The intro-operative blood loss, operating time and complications were recorded. The radiological measurements included preoperative and postoperative spinopelvic parameters, which including thoracic kyphosis (TK), lumbar lordosis (LL), sagittal vertical axis (SVA), pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS) and segmental kyphosis. The recovery of neurological function was evaluated by Frankle classification.Results:All patients were followed up for 7-72 months. Comparing with the cases underwent PVCR technique, the DCVO group has a significantly lower blood loss (1315.00±462.57 ml), operating time (293.00±83.86 min) and complications rate (1.5%). At the time of preoperation, postoperation and last follow-up, the deformity angle of DCVO group was 96.80°±6.32°, 29.10°±6.96°and 29.05°±6.49°, which gained an average 69.9% correction rate. The statistical analysis suggested that deformity angle was enormously corrected. And there was an insignificant difference between DCVO group and PVCR group. Meanwhile, the preoperative, postoperative and follow-up TK of DCVO group was 96.96°±29.13°, 37.15°±4.88° and 37.00°±3.89°respectively, whosecorrection rate was 67.1%; LL was 66.70°±21.21°, 42.25°±5.53° and 41.90°±4.98°, which have a significant difference between pre-operation and post-operation/follow-up ( F=23.997, P<0.001) ; SVA was 75.95±18.63 mm, 16.30±6.88 mm and 16.55±7.30 mm. PI was 47.50°±6.12°, 47.35°±5.54°and 47.90°±5.93°, PT was 37.25°±9.63°, 18.50°±1.99° and 19.00°±1.65°; SS was 10.25°±8.27°, 29.15°±5.91° and 28.85°±5.77°. The sagittal and spinopelvic parameters of two groups improved significantly at postoperation and follow-up. No obviously difference of spinal parameters was found between two groups at preoperation and postoperation. Both groups have cases with dysneuria. And all of these cases achieved different degrees of recovery at follow-up. Conclusion:The use of DCVO technique for the treatment of post-tubercular angular kyphosis is safe and efficiency. DCVO leads a better clinical outcomes and lower complication rate than VCR technique.

Long-term primary culture of mammalian cells has been always difficult due to unavoidable senescence. Conventional methods for generating immortalized cell lines usually require manipulation of genome which leads to change of important biological and genetic characteristics. Recently, conditional reprogramming (CR) emerges as a novel next generation tool for long-term culture of primary epithelium cells derived from almost all origins without alteration of genetic background of primary cells. CR co-cultures primary cells with inactivated mouse 3T3-J2 fibroblasts in the presence of RHO-related protein kinase (ROCK) inhibitor Y-27632, enabling primary cells to acquire stem-like characteristics while retain their ability to fully differentiate. With only a few years' development, CR shows broad prospects in applications in varied areas including disease modeling, regenerative medicine, drug evaluation, drug discovery as well as precision medicine. This review is thus to comprehensively summarize and assess current progress in understanding mechanism of CR and its wide applications, highlighting the value of CR in both basic and translational researches and discussing the challenges faced with CR.

Overlap extension PCR is a common method for site-directed mutagenesis. As objective gene sequence growing longer, it is often difficult to obtain the target product in the second round of PCR, and it is highly possible to introduce unexpected mutations into a long gene fragment by PCR. To circumvent these problems, we can only amplify a small gene fragment which contain the target mutation by overlap extension PCR, and then ligate it with vector to get target plasmid. If the restriction site at the end of the amplified fragment was not a single one on plasmid vector, double fragments ligation method could be used to construct target plasmid. Partial amplification, combined with double fragments ligation, could solve lots of problems in long gene mutagenesis. Taking retinoblastoma gene 1 S780E mutagenesis as an example, it is difficult to amplify whole retinoblastoma gene 1 by overlap extension PCR because of long fragment interfering the overlapping extension of second round PCR. However, it is relatively easy to amplify the F3 (1 968-2 787) fragment which contains target mutation S780E. There is a Nhe I site which can be used for ligation on 5' end of F3 fragment, but another Nhe I site on the plasmid restrained from doing so directly. In order to circumvent this obstacle, we ligated F3 fragment, combining with F2 (900-1 968) fragment which was digested from wild type plasmid, with the vector which contain F1 (1-900) fragment of the gene. That double fragments ligated with one vector at the same time, though less efficient, can recombine into a complete plasmid. The sequences of the two selected recombinant plasmids were consistent with the target mutation, which verified the feasibility of this scheme. As an improvement of overlap extension PCR, partial amplification and double fragments ligation methods could provide solutions for site directed mutagenesis of many long genes.
Base Sequence ; Cloning, Molecular ; Genetic Vectors ; genetics ; Mutagenesis, Site-Directed ; methods ; Nucleic Acid Amplification Techniques ; Plasmids ; Polymerase Chain Reaction

Tumor cells along with a small proportion of cancer stem cells exist in a stromal microenvironment consisting of vasculature, cancer-associated fibroblasts, immune cells and extracellular components. Recent epidemiological and clinical studies strongly support that vitamin D supplementation is associated with reduced cancer risk and favorable prognosis. Experimental results suggest that vitamin D not only suppresses cancer cells, but also regulates tumor microenvironment to facilitate tumor repression. In this review, we have outlined the current knowledge on epidemiological studies and clinical trials of vitamin D. Notably, we summarized and discussed the anticancer action of vitamin D in cancer cells, cancer stem cells and stroma cells in tumor microenvironment, providing a better understanding of the role of vitamin D in cancer. We presently re-propose vitamin D to be a novel and economical anticancer agent.