[Reliability study on establishing a rat tuberculosis wound model using Bacillus Calmette-Guérin].

Objective: To evaluate the reliability of a rat tuberculous wound model established by injecting Bacillus Calmette-Guérin (BCG). Methods: The experimental research was conducted. According to the random number table, fifteen 6-week-old male Sprague-Dawley rats were divided into normal control group and infection group, with 3 rats in normal control group and 12 rats in infection group. Rats in infection group were injected with Freund's complete adjuvant, 3 weeks later, they were injected subcutaneously with BCG bacterial solution to establish a model of tuberculous wounds in rats; rats in normal control group did not receive any treatment. On the 8th, 15th, 32nd, and 43rd day of infection, the skin condition at the injection sites of the rats in infection group was observed roughly. Skin tissue at the injection sites of 3 rats in infection group at each corresponding time point stated above and skin tissue at the corresponding sites of the rats in normal control group were stained with hematoxylin-eosin to observe the cell arrangement, necrosis and inflammation. On 43rd day of infection, acid-fast staining was performed on the skin tissue at the injection sites of the rats in infection group to observe the distribution of bacteria. Results: On the 8th, 15th, 32nd, and 43rd day of infection, tuberculous wound lesions were gradually developed at the skin tissues at the injection sites of the rats in infection group. The cells of the diseased tissue of the rats in infection group arranged disorderly or concentrically, and the number of granulomas and necrotic cells gradually increased, while the skin tissue cells in the corresponding parts of the rats in normal control group arranged regularly with no inflammatory cell infiltration. On the 43rd day of infection, a large number of rod-shaped bacteria were observed in the skin tissue at the injection sites of the rats in infection group. Conclusions: The rat tuberculous wound model established using BCG is stable and reliable, which can meet the experimental requirements.

[Effects of Freund's complete adjuvant on autophagy protein expression in rat tuberculous wound model].

Objective: To explore the effects of Freund's complete adjuvant on autophagy protein expression in rat tuberculous wound model. Methods: The experimental research method was used. In the first batch, twelve 6-week-old male Sprague-Dawley (SD) rats were sensitized by subcutaneous injection of Freund's complete adjuvant into the hips. Three weeks later, the rats were infected with attenuated Bacille Calmette-Guérin (BCG) subcutaneously on both sides of the back spine. After establishing the tuberculosis wound rat model, according to the random number table (the same grouping method below), the rats were divided into 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group, with 3 rats in each group, with continuous normal feeding to the corresponding days after infection. In the second batch, twenty-three 6-week-old male SD rats were divided into blank control group (n=3, normal feeding without any treatment), BCG alone group (n=5), BCG+ rapamycin group (n=6), BCG+ 3-methyladenine group (n=6), and BCG+ starvation group (n=3). The last 4 groups of rats were sensitized as before, and infected as before 1 week later. Rats in BCG alone group were fed normally without any treatment. Rats in BCG+ rapamycin group or BCG+ 3-methyladenine group were intraperitoneally injected with rapamycin or 3-methyladenine once every other day and fed normally. Rats in BCG+ starvation group were fasted for 48 hours after infection and then fed normally. All the rats in the first batch of 4 groups were sacrificed on the corresponding days after infection, and the tissue where the buttocks were injected with Freund's complete adjuvant was harvested; the tissue of rats in the second batch of BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were harvested the same as before 7 days after infection, and all the rats in blank control group were taken the same tissue at the same time point. Hematoxylin-eosin staining was performed to observe the structure and morphology of cells in the tissue harvested; immunohistochemistry was used to observe the protein expressions of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B) in the tissue harvested. Data were statistically analyzed with Kruskal-Wallis test and Bonferroni correction. Results: Inflammatory cell infiltration was observed in the tissue of rats where the Freund's complete adjuvant was injected in 8 d infection group, granuloma formation was seen in 15 d infection group, part of tissue cell necrosis was seen in 32 d infection group and 43 d infection group, and cell necrosis in 43 d infection group was worse than that in 32 d infection group. Seven days after infection, inflammatory cell infiltration was seen in the tissue of rats where the Freund's complete adjuvant was injected in BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group, while regular arrangement of cells and no inflammatory cell infiltration were observed in blank control group. There were no statistically significant differences in the protein expressions of Beclin-1 or LC3B in the tissue of rats where the Freund's complete adjuvant was injected in 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group (H=1.923, 5.821, P>0.05). Seven days after infection, the protein expressions of Beclin-1 and LC3B in the tissue of rats where the Freund's complete adjuvant was injected in blank control group, BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were respectively 0.325% (0.250%, 0.360%), 3.225% (1.340%, 3.987%), 4.823% (2.630%, 6.559%), 4.216% (1.790%, 5.969%), 1.765% (0.865%, 2.649%), and 0.301% (0.264%, 0.516%), 2.865% (1.455%, 5.768%), 1.033% (0.398%, 1.873%), 1.168% (0.429%, 1.907%), 0.655% (0.283%, 1.652%). The protein expression of Beclin-1 in the tissue of rats where the Freund's complete adjuvant was injected in BCG+ rapamycin group was significantly higher than that of blank control group (Z=4.796, P<0.05). The protein expression of LC3B in the tissue of rats where the Freund's complete adjuvant was injected in BCG alone group was significantly higher than that of blank control group (Z=4.953, P<0.05). Conclusions: Freund's complete adjuvant can enhance the expression levels of local tissue autophagy-related proteins Beclin-1 and LC3B in rat tuberculous wound model.

Lesion-aware convolutional neural network for chest radiograph classification.

AIM: To investigate the performance of a deep-learning approach termed lesion-aware convolutional neural network (LACNN) to identify 14 different thoracic diseases on chest X-rays (CXRs). MATERIALS AND METHODS: In total, 10,738 CXRs of 3,526 patients were collected retrospectively. Of these, 1,937 CXRs of 598 patients were selected for training and optimising the lesion-detection network (LDN) of LACNN. The remaining 8,801 CXRs from 2,928 patients were used to train and test the classification network of LACNN. The discriminative performance of the deep-learning approach was compared with that obtained by the radiologists. In addition, its generalisation was validated on the independent public dataset, ChestX-ray14. The decision-making process of the model was visualised by occlusion testing, and the effect of the integration of CXRs and non-image data on model performance was also investigated. In a systematic evaluation, F1 score, sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) metrics were calculated. RESULTS: The model generated statistically significantly higher AUC performance compared with radiologists on atelectasis, mass, and nodule, with AUC values of 0.831 (95% confidence interval [CI]: 0.807-0.855), 0.959 (95% CI: 0.944-0.974), and 0.928 (95% CI: 0.906-0.950), respectively. For the other 11 pathologies, there were no statistically significant differences. The average time to complete each CXR classification in the testing dataset was substantially longer for the radiologists (∼35 seconds) than for the LACNN (∼0.197 seconds). In the ChestX-ray14 dataset, the present model also showed competitive performance in comparison with other state-of-the-art deep-learning approaches. Model performance was slightly improved when introducing non-image data. CONCLUSION: The proposed LACNN achieved radiologist-level performance in identifying thoracic diseases on CXRs, and could potentially expand patient access to CXR diagnostics.

Knockdown of FBP1 enhances radiosensitivity in prostate cancer cells by activating autophagy.

For patients with clinically early-stage localized prostate cancer, radiotherapy is another treatment that can achieve radical treatment in addition to radical prostatectomy. Despite this, there are still a large number of patients with prostate cancer who have a biochemical recurrence after undergoing radiotherapy, or even clinical recurrence, leading to treatment failure. Although the expression of the fructose-1,6-bisphosphatase 1 (FBP1) gene has been found to be absent in various tumors and is associated with a poor prognosis in tumor patients. However, the expression and role of FBP1 in prostate cancer are not clear. The purpose of this study was to investigate the role and mechanism of FBP1 in the radiotherapy resistance of prostate cancer. By analyzing the microarray data of prostate cancer radiotherapy resistant cells and parental cells (GSE53902), we found that FBP1 expression in DU145 radiotherapy resistant cells was significantly higher than in the DU145 parental cells. In addition, we searched for the expression of FBP1 in 492 prostate cancer samples from TCGA and found that its expression in prostate cancer was significantly higher than that in normal tissues. Knockdown of FBP1 expression significantly inhibited the proliferation of prostate cancer cells, promoted DNA damage-mediated apoptosis, and enhanced the sensitivity of prostate cancer cells to radiotherapy. Further mechanism analysis revealed that FBP1 knockdown could activate autophagy mediated by the AMPK-mTOR signaling pathway, while inhibition of the AMPK-mTOR signaling pathway could reverse FBP1 knockdown-mediated autophagy and apoptosis, as well as radiosensitization. In conclusion, this study clarified that FBP1 is an oncogene in prostate cancer, and the main mechanism for knockdown of FBP1 to increase radiosensitivity is to enhance autophagy mediated by the AMPK-mTOR signaling pathway. Therefore, FBP1 may be a potential target for enhancing prostate cancer radiotherapy.

Differential effects of slow rewarming after cerebral hypothermia on white matter recovery after global cerebral ischemia in near-term fetal sheep.

It is widely believed that rewarming slowly after therapeutic hypothermia for hypoxic-ischemic (HI) encephalopathy can improve outcomes, but its impact on white matter injury after HI is unclear. Fetal sheep (0.85 gestation) received 30 min ischemia-normothermia (n = 8), or hypothermia from 3-48 h with rapid spontaneous rewarming over 1 h (ischemia-48 h hypothermia, n = 8), or 48 h with slow rewarming over 24 h (ischemia-slow rewarming, n = 7) or 72 h with rapid rewarming (ischemia-72 h hypothermia, n = 8). Ischemia was associated with loss of total and mature oligodendrocytes and reduced area fraction of myelin basic protein (MBP) and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase; immature/mature oligodendrocytes) and increased microglia and astrocytes. Total numbers of oligodendrocytes were increased by all hypothermia protocols but only ischemia-72 h hypothermia attenuated loss of mature oligodendrocytes. All hypothermia protocols similarly increased the area fraction of MBP, whereas there was only an intermediate effect on the area fraction of CNPase. Microglia were suppressed by all hypothermia protocols, with the greatest reduction after ischemia-72 h hypothermia, and an intermediate effect after ischemia-slow rewarming. By contrast, induction of astrocytes was significantly reduced only after ischemia-slow rewarming. In conclusion, slow rewarming after hypothermia did not improve oligodendrocyte survival or myelination or suppression of microgliosis compared to fast rewarming, but modestly reduced astrocytosis.

Ischemia-reperfusion injury in an aortic dissection patient.

Aortic dissection is a life-threatening emergency. Well-established risk factors include systemic hypertension, hereditary connective tissue diseases (Marfan syndrome and Ehlers-Danlos syndrome), coarctation of the aorta, bicuspid aortic valve, aortitis, and arch hypoplasia. Ischemia of the viscera, the kidneys, the spinal cord, or the lower extremities due to malperfusion constitutes life-threatening complications that have to be considered in the treatment strategy.We report a rare case of symptomatic ischemia of the lower extremities due to aortic dissection. This case demonstrates that the treating physician needs to be vigilant for ischemia reperfusion injuries such as osteofascial compartment syndrome and acute renal failure in aortic dissection.

Initial experiences with laparoscopy and flexible ureteroscopy combination pyeloplasty in management of ectopic pelvic kidney with stone and ureter-pelvic junction obstruction.

To demonstrate the safety and efficacy of combine laparoscopy and flexible ureteroscopy to treat ectopic pelvic kidneys with ureteropelvic junction obstruction (UPJO) and stones. 16 patients of ectopic pelvic kidneys with ureteropelvic junction obstruction and stones were treated with laparoscopy and flexible ureteroscopy (FURS). The operative time, required dose of tramadol, visual analog pain scale (VAPS), postoperative day, stone-free rates (SFRs), perioperative complications, and serum creatinine were evaluated. The SFRs were evaluated with noncontrasted renal computed tomography (CT). Intravenous pyelography (IVP) and CT scan were used to evaluate the UPJO. Stone-free status was defined as absence of stone fragments in kidney or the size of that is less than 3 mm. Operation time from 118 to 225 min, average time (171 ± 28) min; lithotomy time from 16 to 45 min, average time (32 ± 6) min. Average tramadol required at the first day postoperation was (118 ± 49.6) mg; at the second day was (78 ± 24.8) mg. VAPS score at 24 h (5.0 ± 0.7), VAPS score at 48 h (2.5 ± 0.8). Postoperative day (3.9 ± 0.6) days. Stone-free rate was 100%. Average serum creatinine was (88.7 ± 24.3) mol/L before surgery and (92.8 ± 21.6) mol/L after surgery. No major complication. No stone and obstruction recurrence in the follow-up of average 29.3 months. Combined FUR and LC is a good option for patient of ectopic pelvic kidney with renal stone and UPJO. From our initial experience, the SFRs and the effect of pyeloplasty are satisfactory and without major complication, the operative time is acceptable.

A new subtype of crossed fused ectopia of the kidneys.

Crossed fused renal ectopia is a rare congenital anomaly; here, we report a new subtype of crossed fused renal ectopia associated with the retroiliac megaureter and thoracic scoliosis deformity. It is beyond the traditional classification of crossed fused renal ectopia. There are 2 kidneys in the left and hydronephrosis of the upper kidney, the right kidney crossed over and fused with the lower kidney of the left. It is never seen in previous reports. Recurrent infection was cured by resecting the hydronephrosis of the upper kidney and retroiliac megaureter.

Genitourinary toxicity of ketamine.

Ketamine is a relatively new recreational drug used by youngsters in recent decades. Its toxic effects on the genitourinary system were first reported in 2007, and now attract extensive attention from urologists, pharmacologists, and toxicologists all over the world. As many front-line health professionals and medical social workers are still unaware of this new clinical entity and an increasing number of the drug users seek help for urological symptoms, this mini-review aimed to summarise the clinical features and possible mechanisms of ketamine-induced genitourinary toxicity. By raising public awareness of these toxic effects, the authors hope that the contents of this review will be widely disseminated not only to medical professionals, but also to relevant government departments and the general public.

Salicylate, an aspirin metabolite, specifically inhibits the current mediated by glycine receptors containing alpha1-subunits.

BACKGROUND AND PURPOSE: Aspirin or its metabolite sodium salicylate is widely prescribed and has many side effects. Previous studies suggest that targeting neuronal receptors/ion channels is one of the pathways by which salicylate causes side effects in the nervous system. The present study aimed to investigate the functional action of salicylate on glycine receptors at a molecular level. EXPERIMENTAL APPROACH: Whole-cell patch-clamp and site-directed mutagenesis were deployed to examine the effects of salicylate on the currents mediated by native glycine receptors in cultured neurones of rat inferior colliculus and by glycine receptors expressed in HEK293T cells. KEY RESULTS: Salicylate effectively inhibited the maximal current mediated by native glycine receptors without altering the EC(50) and the Hill coefficient, demonstrating a non-competitive action of salicylate. Only when applied simultaneously with glycine and extracellularly, could salicylate produce this antagonism. In HEK293T cells transfected with either alpha1-, alpha2-, alpha3-, alpha1beta-, alpha2beta- or alpha3beta-glycine receptors, salicylate only inhibited the current mediated by those receptors that contained the alpha1-subunit. A single site mutation of I240V in the alpha1-subunit abolished inhibition by salicylate. CONCLUSIONS AND IMPLICATIONS: Salicylate is a non-competitive antagonist specifically on glycine receptors containing alpha1-subunits. This action critically involves the isoleucine-240 in the first transmembrane segment of the alpha1-subunit. Our findings may increase our understanding of the receptors involved in the side effects of salicylate on the central nervous system, such as seizures and tinnitus.