Objective To understand the infectious pathogen characteristics and drug sensitivity of hospitalized patients in the respiratory intensive care unit (RICU) of Renmin Hospital of Wuhan University. Methods Bacterial culture samples sent to the RICU of our hospital from January 2020 to December 2022 were retrospectively analyzed. The bacterial types were identified by Bruker mass spectrometer, and the Phoenix 100 was used for drug sensitivity analysis. The antimicrobial susceptibility was analyzed by WHONET 5.6 software. Results A total of 1 157 strains of bacteria were isolated, including 878 strains of Gram-negative bacteria (75.89%) and 279 strains of Gram-positive bacteria (24.11%). The top five with the highest detection rate were Acinetobacter baumannii (25.50%), Pseudomonas aeruginosa (18.76%), Klebsiella pneumoniae (13.83%), Staphylococcus aureus (6.57%) and Escherichia coli (5.70%). Among them, Acinetobacter baumannii was extremely drug-resistant, only showing relatively high sensitivity to colistin, minocycline, and tigecycline. Staphylococcus aureus accounted for the highest proportion of Gram-positive bacteria (6.57%), with methicillin-resistant Staphylococcus (MRSA) showing a continuous increase. Conclusion In the past three years, Gram-negative bacteria have been the main pathogenic bacteria detected in the respiratory intensive care unit of our hospital. The main bacteria are Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae, which have a high resistance rate to various antibiotics. Therefore, clinical monitoring of resistant strains in RICU should be strengthened to facilitate rational use of antibiotics and improve antibacterial effect.

Objective This study aims to establish a non-puerperal mastitis (NPM) rat model by simulating hyperprolactinemia and immune-inflammatory states, and to evaluate its local inflammatory characteristics in the mammary gland, thereby laying the foundation for research on the diagnosis and treatment of this clinically challenging disease. Methods Twelve SPF-grade Wistar female rats were evenly divided into a control group and a model group. During the experiment, the control group received no experimental treatment or medication. The model group received daily subcutaneous injections of 100 mg/kg metoclopramide hydrochloride for 7 consecutive days. Serum prolactin (PRL) levels were measured using ELISA on the 10th, 20th, and 30th days after the first injection. After 7 days of injections, 200 μL of lactating SD rat milk was mixed with 200 μL of complete Freund's adjuvant to prepare an oil-in-water emulsion, which was administered by multiple subcutaneous injections into the back of the Wistar rats for the initial immunization. Seven days after the initial immunization, the emulsion was injected subcutaneously into the third, fourth, and fifth mammary glands for the final immunization. After the final immunization, the rats were observed for 28 days for changes in mammary gland appearance, and the size of mammary nodules was calculated. On the 3rd, 7th, 14th, and 28th days, hematoxylin-eosin (HE) staining was used to analyze mammary tissue morphology. Immunohistochemistry was employed to detect CD138 expression levels. ELISA was used to measure the levels of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, and inducible nitric oxide synthase (iNOS) in mammary tissue to comprehensively assess the model. Results Rats in the model group exhibited mammary skin ulceration and foul odor at the ulcer sites. Palpation and ultrasound revealed the formation of mammary nodules. HE staining showed that on the 3rd day after the final immunization, normal ductal and lobular structures in the mammary glands disappeared, with significant infiltration of plasma cells. On the 7th day, ductal dilation, epithelial necrosis and detachment, and pronounced periductal plasma cell and lymphocyte (predominantly T-lymphocytes) infiltration were observed. On the 14th day, there was a proliferation of fibrofatty tissue, small blood vessels, and granulation tissue, with scattered plasma cells in the interstitium. By the 28th day, inflammatory cell infiltration and fibrous tissue proliferation were reduced, with granuloma formation. Serum PRL levels in the model group were significantly increased on the 10th day (P<0.05) and the 20th day (P<0.001). IL-6 and TNF-α levels in mammary tissue were higher in the model group compared to the control group on the 3rd, 7th, 14th, and 28th days (P<0.05). IL-1β levels were higher on the 3rd, 7th, and 14th days compared with the control group (P<0.01) but lower than the control group on the 28th day (P>0.05). iNOS levels were significantly higher on the 7th day after the final immunization (P<0.001). Conclusion A successful NPM model was established in rats, which exhibited typical pathological features such as local mammary masses, abscesses, ulcers, ductal dilation and plasma cell infiltration. This model can serve as a foundation for further research into the diagnosis and treatment of this clinically challenging disease.

Background::Bladder cancer, characterized by a high potential of tumor recurrence, has high lifelong monitoring and treatment costs. To date, tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types. Nonetheless, the existence of soft tumor cells in bladder tumors remains elusive. Thus, our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods::The stiffness of bladder cancer cells was determined by atomic force microscopy (AFM). The modified microfluidic chip was utilized to separate soft cells, and the 3D Matrigel culture system was to maintain the softness of tumor cells. Expression patterns of integrin β8 (ITGB8), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) were determined by Western blotting. Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59 (TRIM59). The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models. Results::Using our newly designed microfluidic approach, we identified a small fraction of soft tumor cells in bladder cancer cells. More importantly, the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens, in which the number of soft tumor cells was associated with tumor relapse. Furthermore, we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells. Simultaneously, we detected a remarkable up-regulation in ITGB8, TRIM59, and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions::The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness. Meanwhile, the soft tumor cells become more sensitive to chemotherapy after stiffening, that offers new insights for hampering tumor progression and recurrence.

Objective : To investigate the expression of programmed death 1 (PD1) on CXCR5 －CD4 + T cells from the patients with rheumatoid arthritis (RA) and to analyze the clinical relevance to disease severity． Methods : The expression of PD1 on CXCR5 －CD4 + T cells were examined from 82 RA patients and 46 healthy controls (HC) by the technique of flow cytometry．The expression of PD1 including mean fluorescence intensity (MFI) on CXCR5 - CD4 + T cells and percentage of PD1 + CXCR5 －CD4 + T cells were compared between RA patients and HC．Moreo- ver，its correlation with laboratory inspection was analyzed. Results : ① The percentage of PD1 + CXCR5 －CD4 + T cells and the MFI of PD1 on CXCR5 －CD4 + T cells from RA patients were significantly elevated compared with HC (P＜0. 000 1，U = 1 082 ; P ＜0. 000 1 ，U = 917. 5) . ② The percentage of PD1 + CXCR5 － CD4 + T cells in RA patients were positively associated with rheumatoid factors (RF) (rs = 0. 267 9，P = 0. 022 9) ，increased RF (rs = 0. 419，P = 0. 001 1) ，increased anti-cyclic citrullinated peptide (anti-CCP) (rs = 0. 260 7，P = 0. 040 7) ，the percentage of plasmablasts (rs = 0. 302 4，P = 0. 029 3) ，DAS28-ESR (rs = 0. 244 7，P = 0. 042 7) . ③ The MFI of PD1 on CXCR5 －CD4 + T cells in RA patients were negatively with lymphocytes number (rs = －0. 338 7，P = 0. 002 0 ) ，lymphocytes percentage ( rs = －0. 347 9 ，P = 0. 001 4 ) ，lymphocyte-to-monocyte ratio ( rs = - 0. 263 7，P = 0. 017 4) ．The MFI of PD1 on CXCR5 － CD4 + T cells in RA patients were positively associated with neutrophil percentage ( rs = 0. 304 7 ，P = 0. 005 4 ) ，neutrophilto-lymphocyte ratio ( rs = 0. 341 1 ，P = 0. 001 8) ，platelet-to-lymphocyte ratio (rs = 0. 232 1，P = 0. 037 1) ，systemic immune inflammation index (rs = 0. 288 0，P = 0. 009 1) ，derived Neutrophil to lymphocyte ratio ( rs = 0. 320 3，P = 0. 003 6) ，erythrocyte sedi- mentation rate (rs = 0. 259 5，P = 0. 019 3) ，patient visual analogue scale (rs = 0. 241 6，P = 0. 043 9) ．The MFI of PD1 on CXCR5 － CD4 + T cells from high active RA patients was significantly elevated compared with non-high active group (P = 0. 040 7，U = 406. 0) ． Conclusion The abnormal expression of PD1 on CXCR5 －CD4 + T cells are observed in patients with RA．Increased expression of PD1 on CXCR5 － CD4 + T cells are associated with the production of antibody，the percentage of plasmablasts，inflammation，and also disease activity.

Objective : To investigate the expression and clinical significance of erbb2 interacting protein (ERBIN) and PDZ domain containing 2 protein (PDZD2) in colorectal cancer tissues，as well as the effects of ERBIN and PDZD2 on the progression of colorectal cancer and the corresponding mechanisms． Methods : Western blot was used to detect the expression of ERBIN and PDZD2 proteins in tumor tissues and corresponding adjacent tissues from 86 colorectal cancer patients，as well as normal human colon fibroblasts ( CCD-18Co) and human colorectal cancer cell lines ( SW480 ，HCT116 ，SW620 and HT29 ) ; SW620 and HCT116 cells were transfected with ERBIN or PDZD2 overexpression vector，and then cell proliferation，cell invasion and apoptosis were detected．The expres- sion of ERBIN or PDZD2 protein in SW620 cell was observed by immunofluorescence chemistry．Co-immunoprecip- itation assay was used to detect the interaction between ERBIN and PDZD2 proteins. Results : The expression levels of ERBIN and PDZD2 proteins in colorectal cancer tissues were lower than those in adjacent tissues (P＜0．01) ， and the expression levels of ERBIN and PDZD2 in colorectal cancer cells were lower than those in CCD-18Co cells (P＜0. 01) ．The expression levels of ERBIN and PDZD2 proteins were correlated with the depth of invasion，dif- ferentiation，TNM staging and lymph node metastasis of colorectal cancer (P＜0. 001) ; ERBIN or PDZD2 overex- pression reduced the proliferation and invasion of SW620 and HCT116 cells (P＜0. 01) ，increased cell apoptosis (P ＜0. 01 ) ; ERBIN directly bound to PDZD2 ，and overexpression of ERBIN increased the expression level of PDZD2 protein． Conclusion ERBIN can inhibit the proliferation and invasion of colorectal cancer cells and pro- mote apoptosis by promoting the expression of PDZD2 protein.

Objective:To investigate the clinical features, treatment and prognosis of anti-γ-aminobutyric acid type B receptor (GABA B R) encephalitis. Methods:Retrospective analysis of five patients of anti-GABA BR encephalitis from the Department of Neurology, the University of Hong Kong-Shenzhen Hospital from September 2017 to June 2019 was carried out. Clinical manifestations, auxiliary examination, and treatment were analyzed. The patients were followed up for 3.5-23.0 months to assess their prognosis. Results:Five cases of anti-GABA BR encephalitis (19-81 years old) presented acute onset, with refractory epilepsy as the main clinical manifestation. There were hyperintensive signals on T 2/fluid attenuated inversion recovery in four patients′ temporal lobe and hippocampus. Electroencephalogram showed slow wave or epileptic discharge; Lung mass was found in four patients, and all were small cell lung cancer. Five cases had poor response to first-line immunotherapy (intravenous use of pulse methylprednisolone, high dose immunoglobulin or plasma exchange), then three patients received second-line immunotherapy (rituximab, cyclophosphamide), two of whom with tumor also received tumor chemotherapy. Patients who received second-line treatment and tumor chemotherapy showed better outcome than those who only received first-line treatment. Conclusions:Anti-GABA BR encephalitis present with limbic encephalitis syndromes characterized by refractory epilepsy. For patients with poor response to first-line immunotherapy, initiating second-line immunotherapy as soon as possible can improve the prognosis significantly.

Objective To investigate whether mild hypothermia can promote neurogenesis in the dentate gyrus of hippocampus and cognitive function recovery after traumatic brain injury ( TBI) through inhibiting apoptosis of hippocampal neurons. Methods A total of 66 healthy adult Sprague-Dawley rats were randomly divided into sham group, TBI group and TBI+hypothermia group, with 22 rats in each group. The rat TBI model was established using the fluid percussion device. The rats in TBI +hypothermia group received 4-hour hypothermia therapy immediately after injury, with the target temperature of 33. 5℃. Bromodeoxyuridine (BrdU) was injected into the rats' abdominal cavity to label the mitotic cells. The test of Morris water maze was used to evaluate the rats' spatial learning and memory capabilities. Immunofluorescence staining was used to observe the expression levels of BrdU, doublecortin (DCX), neuron specific nuclear protein (NeuN), cysteinyl aspartate specific proteinase 3 (caspase-3) and cleaved caspase-3 expressions in dentate gyrus of hippocampus at 7 days and 28 days after injury. Expressions apoptosis-related proteins including the factor associated suicide ( FAS )/factor associated suicide ligand (FASL), B-cell lymphoma-2 (Bcl-2), caspase-3 and cleaved caspase-3 expressions were detected by Western blot assay. Results The water maze tests at 28 days after injury showed that compared with TBI group, the escape latency in TBI+hypothermia group was significantly shorter [(24. 2 ± 5. 9)s:(18 ± 4. 1)s], and both the time in the target quadrant and the number of platform crossing were increasedsignificantly[(24.9±6.5)s:(31.7±5.2)s; (1.9±0.8) times:(3.5±1.2)times](P<0. 05). Compared with the sham group, in TBI group and TBI+hypothermia group, the BrdU+ new-born cells in the dentate gyrus of hippocampus were significantly increased at 7 days after injury [(9. 4 ± 4. 1):(33. 4 ± 3. 8);(9. 4 ± 4. 1):(45. 8 ± 5. 6)], the BrdU+ /DCX+ new-born neurons were increased at 7 days after injury [(2. 0 ± 0. 6):(9. 6 ± 1. 6);(2. 0 ± 0. 6):(19. 2 ± 3. 7)], and the BrdU+ /NeuN+mature neurons were increased at 28 days after injury [(2. 6 ± 1. 0) :(17. 2 ± 3. 9); (2. 6 ± 1. 0) :(33. 6 ± 9. 1)] (P<0. 01). TBI group showed more obvious increase than the TBI+hypothermia group (P<0. 01). Moreover, compared with 7 days after injury, the number of BrdU+ cells at 28 days after injury was further increased in TBI +hypothermia group but decreased in TBI group [(45. 8 ± 5. 6) :(58. 8 ± 9. 2);(33. 4 ± 3. 8):(22. 0 ± 3. 5)](P<0. 05 or <0. 01). Compared with the sham group, the caspase-3 +NeuN+ and caspase-3 +NeuN+ apoptotic neurons were significantly increased at 7 days after injury in TBI group [(2. 0 ± 0. 9):(11. 6 ± 2. 6); (2. 6 ± 1. 0):(10. 2 ± 2. 9)] (P<0. 05). Compared with the TBI group, the cleaved caspase-3 +NeuN+ apoptotic neurons were decreased in TBI+hypothermia group [(6. 6 ± 2. 0):(11. 6 ± 2. 6)](P<0. 05). Furthermore, compared with the TBI group, mild hypothermia might down-regulate the expression of FAS, FASL, cleaved caspase-3 and caspase-3 and up-regulate the expression of Bcl-2 in the hippocampus [(1. 54 ± 0. 15) :(1. 14 ± 0. 12);(1. 06 ± 0. 04):(0. 80 ± 0. 09); (0. 84 ± 0. 03):(0. 62 ± 0. 08); (0. 93 ± 0. 06):(0. 86 ± 0. 09);(0. 71 ± 0. 01):(1. 58 ± 0. 18)](P<0. 05). Conclusions Mild hypothermia might inhibit apoptosis of hippocampal neurons through cleaved caspase-3, FAS/FASL and Bcl-2 pathways, thus improving the neurogenesis and maturation of neurons in the dentate gyrus of hippocampus and facilitating cognitive function recovery in rats. It indicates that the function of hypothermia in anti-apoptosis and neurogenesis and maturity of hippocampal neurons may have a potential role in predicting the prognosis of TBI patients.

Objective: To compare the efficacy and safety of mycophenolate mofetil versus cyclosporine A in treating children with primary refractory nephrotic syndrome. Methods: Conducted a prospective randomized controlled clinical trial in 62 pediatric patients (including 44 boys and 18 girls), age ranged from 2.1 to 17.0 years; 32 cases presented with frequently relapsing nephrotic syndrome (FRNS) and 30 cases presented with steroid-resistant nephrotic syndrome (SRNS), who were admitted to department of Nephrology, Children's Hospital Affiliated to Capital Institute of Pediatrics from October 2013 to October 2015. The patients received either mycophenolate mofetil (20-30)mg/(kg·d) or cyclosporine A (3-5)mg/(kg·d) randomly, on the basis of prednisone treatment. Follow-up interview was conducted regularly for at least one year. Efficacy rate, relapse rate, time required for induction of remission, relapse-free period and prednisone dosage were compared between the two groups. Results: (1) Renal histologic examination, which was available for 17 patients, revealed minimal change disease in 8 patients, mesangial proliferative glomerulonephritis (MsPGN) in five, membranous nephropathy in two, and focal segmental glomerulosclerosis (FSGS) in two. (2) Comparison of mycophenolate mofetil versus cyclosporine A in children with FRNS: There were 14 patients with FRNS in mycophenolate mofetil group and 18 patients with FRNS in cyclosporine A group respectively. The relapse rate (episodes/year) in cyclosporine A group was lower than that of mycophenolate mofetil group (1.0 (0.0, 1.0) vs. 1.0 (1.0, 3.0), Z=-2.405, P=0.016). The relapse-free period (months) in cyclosporine A group was longer than that of mycophenolate mofetil group (10.0 (5.7, 12.1) vs. 5.0 (1.0, 11.0), Z=-1.984, P=0.047). No significant difference in dosage of prednisone was found between cyclosporine A and mycophenolate mofetil groups when followed up for 1 year. (3) Comparison of mycophenolate mofetil versus cyclosporine A in children with SRNS: The efficacy rate was 6/14 in mycophenolate mofetil group and 13/16 in cyclosporine A group. The complete remission rate was 4/14 in mycophenolate mofetil group and 12/16 in cyclosporine A group (P<0.05). The time (months) required for induction of remission in cyclosporine A group was significantly shorter than that of mycophenolate mofetil group (1.0 (1.0, 2.0) vs. 3.0 (2.5, 4.0), Z=-2.529, P=0.011). No significant differences were found between the two groups with respect to relapse-free period and relapse rate. (4) Except that one patient developed hypertensive encephalopathy in cyclosporine A group, no other serious adverse events were recorded. There were no significant differences between two groups with respect to adverse events. Conclusion Our results indicated that both mycophenolate mofetil and cyclosporine A were effective in the treatment of children with refractory nephrotic syndrome. Cyclosporine A was superior to mycophenolate mofetil in preventing relapses in patients with FRNS and inducing complete remission in patients with SRNS. Although most patients were able to tolerate mycophenolate mofetil and cyclosporine A, but the toxicity and safety of cyclosporine A should be monitored closely.

BACKGROUND:Currently, morphological observations of brain cavity after traumatic brain injury (TBI) via cadavers or animal specimen are difficult to obtain dynamic changes.
OBJECTIVE:To explore the application effect of MRI-based three-dimensional (3D) reconstruction for evaluating the prognosis of TBI.
METHODS:Five male Sprague-Dawley rats were enrol ed to establish TBI models by Electronic Cortical Contusion Injury (eCCI), and scanned by 3.0T MRI with Rat-coil to obtain the DICOM date of brain at 1 day, 1, 2 and 3 months after modeling. Brain cavities were 3-dimensional y reconstructed by Mimics16.0 software, and analyzed in the Meshmixer software.
RESULTS AND CONCLUSION:(1) The outline of reconstruction model image was clear, and could be observed and measured from different sides and perspectives. (2) The cavity volume and surface area at different time points after TBI showed significant differences between each other except that at 2 and 3 months (P<0.05). (3) The results of cavity change suggested that the cavity tended to be regular after 3 months of TBI. (4) In conclusion, 3D reconstruction software Mimics combining with model analysis software Meshmixer can conveniently and quickly obtain the cavity model, and provide an intuitive way for evaluating the dynamic variations of the brain cavity after TBI.

Objective To investigate the clinical effect of flupentixol and melitracen tablets combined with macrogol 4000 powders on treatment of irritable bowel syndrome with constipation (IBS-C) in elderly patients. Methods Fifty paients aged from 65 to 95 years with IBS-C were randomly divided into 2 groups: control group (23 patients) and treatment group (27 patients). The patients in treatment group received flupentixol and melitracen tablets (1 tablets, once a day) combined with macrogol 4000 powders (1 bag/time, 2 times a day) , while the patients in control group received macrogol 4000 powders alone. The treatment period was 4 weeks. The curative effect, the scores of self-rating depression scale (SDS) and self-rating anxiety scale (SAS) and adverse reaction were compared between two groups. Results At the improvement of depressive symptoms, the effective rate in treatment group and control group was 88.9%(24/27) and 39.1%(9/23), and there was significant difference (P<0.01). At the improvement of anxiety symptoms, the effective rate in treatment group and control group was 74.1%(20/27) and 26.1%(6/23), and there was significant difference (P<0.01). The total effective rate in treatment group and control group was 88.9%(24/27) and 65.2%(15/24), and there was significant difference (P<0.05). Conclusions Flupentixol and melitracen tablets combined with macrogol 4000 powders on treatment of aged IBS-C patients have more distinguished therapeutic effect in improving constipation symptoms, stools times, anxiety and depression.