[Clinical, imaging, and pathological characteristics of 35 cases of mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome].

Objective: To summarize the clinical, imaging, and pathological characteristics of mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS) to improve the diagnosis of this rare disease. Methods: A retrospective case series was conducted to collect the clinical data and results of genetic testing, muscle biopsy, and imaging studies including computed tomography (CT), magnetic resonance imaging (MRI), and magnetic resonance spectroscopy (MRS) of 35 patients with MELAS admitted to the Nanjing Drum Tower Hospital from 2012 to 2021. Descriptive statistical analysis including mean, standard deviation, and frequency percentage were carried out. Results: The average age of onset of the patients was 30.2±2.3 years; the prevalence of family history was 20%. The two main initial symptoms were limb weakness and convulsions. The clinical manifestations of the neuromuscular system were proximal muscle weakness and exercise intolerance. The endocrine system is the most affected outside the neuromuscular system, with diabetes being the most common condition. Among the five patients who underwent brain CT, four showed hypodense lesions and two had calcified lesions. Brain MRI in 26 patients showed that the lesions more often affected the parietal lobe, basal ganglia, temporal lobe, occipital lobe, and frontal lobe than the infratentorial areas. Twelve of these individuals exhibited different levels of brain atrophy. Among the 10 patients who underwent 1H-MRS, nine showed a decrease in N-acetylaspartate (NAA) levels, eight exhibited abnormal lactate elevation (Lac peaks), whereas six had both reduced NAA levels and the presence of Lac peaks. Thirty-one patients underwent genetic testing; among them, 25 were found to have the mt.3243A>G mutation, while the remaining six exhibited rare gene alterations. Muscle biopsies were performed in 21 patients, and 15 showed abnormal mitochondrial proliferation manifested by ragged red fibers and defective oxidative phosphorylation manifested by cytochrome C oxidase (COX) enzyme-deficient muscle fibers. Conclusion: The clinical manifestations of MELAS syndrome are variable and complex, and early atypical symptoms could be missed or misdiagnosed. A detailed clinical history, imaging MRS analysis, muscle biopsy, and genetic testing are necessary to confirm the accurate diagnosis of MELAS.

Experimental Study on Flame Chemical Composition of Coal and Ammonia Gas-Solid Jet in Flat Flame Burner.

Ammonia as a fuel to partially or completely replace fossil fuels is one of the effective ways to reduce carbon dioxide, and the research on ammonia coal cocombustion is of great significance. The combustion characteristics of ammonia are very different from those of pulverized coal, resulting in the ignition and emission characteristics of ammonia and pulverized coal gas flow that is different from traditional pulverized coal flame. In this paper, the effect of pulverized coal concentration in coal and ammonia mixed combustion jet on the ignition distance and gas-phase components at different positions of the jet flame were studied experimentally on the flat flame burner, and the conditions of ignition and ignition stability of coal and ammonia gas-solid fuel were expounded. It was found that the ammonia mixed with pulverized coal changed the temperature field of the flat flame burner and therefore the ignition characteristics of the jet were changed. The ignition delay time at the same jet speed was positively correlated with the pulverized coal concentration, but when the pulverized coal concentration continued to decrease, the influence on the ignition delay time gradually became smaller. The composition of coal ammonia gas-solid fuel changed the heat transfer path and share during combustion, and finally, the flame temperature was negatively correlated with the concentration of pulverized coal. Therefore, the reduction of the pulverized coal concentration was conducive to the stable combustion of coal ammonia mixed fuel. When HAB = 100 mm, the conversion rate of fuel N to NOx per unit mass of coal ammonia mixture increased with the increase of pulverized coal concentration. The NOx production amount first increased and then decreased with the increase of pulverized coal concentration, and the amount of N2O and NO2 decreased rapidly with the increase of HAB. The proportion of NOx in NO exceeded 94%, which was conducive to achieving low nitrogen combustion of coal and ammonia gas-solid fuel. In general, the O2 concentration in the ammonia coal jet flame decreased, the flue gas temperature, and NOx and CO generation increased after mixing ammonia, and the optimal pulverized coal concentration in this experiment was 0.41 kgc/kga (mass ratio of pulverized coal to the sum of N2 and NH3).

[A comparative study on the construction methods of animal models of aortic arch dissection].

Objective: To compare the effectiveness and safety of different methods to construct animal models of aortic arch dissection (AAD), and explore safe and effective methods for constructing AAD animal models. Methods: Twenty-four healthy mongrel dogs were divided into 4 groups by random number table (n=6). Group A: Venous incision needle high pressure water flow impact method; Group B: Venous incision needle non-high pressure water flow impact method; Group C: Transarterial sheath non-high pressure water flow impact method; Group D: Two-way balloon expansion combined with elastase perfusion method. Imaging examinations were performed immediately and 7 days after operation, aortic tissue biopsy and pathological staining were performed 15 days after operation to observe the formation of AAD. The operation time, aortic blood flow block time, model construction success rate, dissection tear length, postoperative survival rate and survival time of four groups of experimental dogs were collected to compare the effectiveness and safety of different construction methods. Results: There were no significant difference of the gender, age and weight between four groups of experimental dogs (all P>0.05). The operation time of four groups of experimental dogs were (111.6±8.0), (168.0±17.4), (164.4±13.9), (202.8±21.5)min, and the difference was statistically significant (F=39.973, P<0.001). The operation time of group A was significantly lower than group B, C and D (all P<0.001). The aortic blood flow block time of four groups of experimental dogs were (5.2±1.8), (19.6±3.8), (20.6±3.9), and (18.6±3.0) min, and the difference was statistically significant (all P<0.001). The aortic blood flow block time of group A was significantly lower than group B, C and D (F=27.598, P<0.001). The four groups of experimental dogs had 5, 5, 4, and 1 model were successfully constructed, respectively, and the difference was statistically significant (P=0.008). The successful rate of model construction in group A was significantly higher than that in group D (P=0.040). The dissection tear length of four groups were (14.4±3.0), (11.3±4.2), (7.0±2.3), (4.7±0.6) cm,and the difference was statistically significant (F=8.103, P=0.003). The dissection tear length of group A was significantly longer than group C, D (all P<0.05). The postoperative survival time were 15.0(10.0, 15.0), 5.0(3.0, 10.0), 3.5(1.5, 4.8), 10.0(2.8, 15.0) days, and the difference was statistically significant (χ2=7.825,P=0.036). The postoperative survival time of group A was significantly higher than group B, C (all P<0.05). There was no significant difference in the survival rate of the four groups (P=1.000). The pathological staining results showed that the elastic fiber at the tearing point of AAD was destroyed, and the elastic fiber on the outer wall of the false cavity was over-stretched, which was consistent with the pathological changes of aortic dissection. Conclusion: Transvenous incision needle high-pressure water flow impact modeling method is easy to operate. The aortic blood flow block time is short, the dissection tear length is wide, and the postoperative survival time is long, can be used as the preferred method of animal AAD model construction.

Apelin-13/APJ system delays intervertebral disc degeneration by activating the PI3K/AKT signaling pathway.

OBJECTIVE: To study the effect of Apelin-13/APJ system on intervertebral disc degeneration and its mechanism. PATIENTS AND METHODS: This study detected the expression of APJ in human intervertebral disc tissue with varying degrees of degeneration. IL-1ß is used to stimulate the degeneration of nucleus pulposus cells. We used recombinant human Apelin-13 and Ala13 to activate and inhibit the APJ receptor, respectively. The inhibitor LY294002 was used to inhibit the PI3K/AKT signaling pathway. We studied the effects of Apelin-13/APJ system on nucleus pulposus cells and its mechanism by Western blot, RT-PCR, and so on. RESULTS: APJ is lowly expressed in the nucleus pulposus of patients with a high degree of degeneration. IL-1ß stimulates the nucleus pulposus cells and reduces the expression of APJ in nucleus pulposus cells. Recombinant human Apelin-13 reduces the degradation of nucleus pulposus extracellular matrix, promotes proliferation, and reduces the levels of apoptosis and inflammation. In addition, the Apelin-13/APJ system increases the expression of PI3K and AKT and activates the PI3K/AKT signaling pathway. CONCLUSIONS: Apelin-13/APJ system activates PI3K/AKT signaling pathway activity, reduces the degradation of nucleus pulposus extracellular matrix, promotes proliferation, and reduces the level of apoptosis and inflammation, thus delaying the degeneration of the intervertebral disc.

[Clinicopathological characteristics, MSI and K-ras gene mutations of double primary malignancies associated with colorectal cancer].

Objective: To investigate the clinicopathological characteristics, MSI and K-ras mutation of double primary malignancies (DPM) associated with colorectal cancer (CRC). Methods: From January 2015 to December 2016, the clinicopathological data of CRC patients treated by surgery in the Affiliated Drum Tower Hospital of Nanjing University Medical School were collected, and the clinical data was analyzed. Multiplex real-time fluorescence quantitative PCR and amplification refractory mutation was performed to identify MSI and K-ras gene mutations. Results: Of all patients with CRC, 5.2% (55/1 066) were DPM. There was no significant difference in the male and female ratio, age, colorectal cancer site, T stage, N stage composition ratio between DPM patients with CRC and patients with single CRC (P>0.05). There were significant difference of TNM stage between the two group (P<0.05). The most frequent location of CRC was the colon in both DPM patients with CRC and patients with single CRC[35.5% (359/1 011) and 41.8% (23/55), respectively]. Of 55 DPM patients with CRC, 48 were metachronous DPM patients, 7 were synchronous DPM patients and 41 were colorectal cancer first. In extracolonic organ, digestive system (23/55) was the most commonly occurring system and stomach (11/55) was the most common lesion. DPM patients with CRC had higher incidence of MSI-H than patients with single CRC (P<0.05). There was no significant difference of K-ras gene mutation between DPM patients with CRC and patients with single CRC (P>0.05). MSI-H and K-ras mutation were present in only 2 patients of DPM patients with CRC. Conclusions: The rectum is the most common lesion site in CRC patients. The stomach is the most common extracolonic organ of DPM patients with CRC. DPM patients with CRC has high risk of MSI-H, but no significant difference in the incidence of K-ras mutation.

Effect miR-214-3p on proliferation and apoptosis of breast cancer cells by targeting survivin protein.

OBJECTIVE: Studies showed that microRNAs (miRs) play an important role in the development of breast cancer. It has been shown that there were significant differences between the expression levels of serum miR-214-3p in breast cancer patients and healthy controls. Since survivin is involved in cell cycle and apoptosis, this study aims to investigate the effect of miR-214-3p on the proliferation and apoptosis of breast cancer cells. MATERIALS AND METHODS: Dual-Luciferase reporter system was used to validate the cell cycle-related target gene survivin. miRanda and TargetScan were used to predict miR-214-3p target genes. Lipofectamine 2000 was used to transfect the miR-214-3p mimics, miR-NC into the MCF-7 cells. The quantitative Real Time-PCR (qRT-PCR) was used to detect the expression levels of miR-214-3p and survivin. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to examine the cell proliferation of breast cancer cells. The flow cytometry assay was used to evaluate the apoptosis of breast cancer cells. RESULTS: Dual-Luciferase reporter assay showed that cells co-transfected with wild-type vector and miR-214-3p mimics had significant lower ratios of hRluc/Luc fluorescence compared to that of the control group (p<0.05). The expression level of miR-214-3p was increased along with the increase of time after transfection, whereas the expression level of survivin mRNA was decreased along with the increase of time post transfection. This result suggests that miR-214-3p regulates the mRNA expression of survivin. Transfection of miR-214-3p inhibitor increased the proliferation of MCF-7 cells and transfection of miR-214-3p mimics decreased the proliferation of MCF-7 cells compared to control group (p<0.05). CONCLUSIONS: Survivin gene is a downstream target of miR-214-3p in breast cancer cells. The expression of miR-214-3p and survivin is correlated with the proliferation and apoptosis of breast cancer cells.

[Visceral parasitic migration of liver: a clinicopathologic analysis of seven cases].

Objective: To investigate the clinical, radiological and pathological features of visceral parasitic migration of the liver. Methods: Seven cases of visceral parasitic migration of liver were identified at the Affiliated Drum Tower Hospital of Medical School of Nanjing University from January 2008 to July 2017. Clinical data, enhanced CT image and pathological features were analyzed, combining with literature review. Results: There were 5 male and 2 female patients. Five patients presented with abdominal pain or discomfort as the first symptom. Two patients were admitted to the hospital for physical examination with liver nodule. Blood eosinophils were mildly to moderately increased in 4 cases. Enhanced CT showed the liver irregular beaded nodules that showed no significant enhancement of arterial phase. Mild enhancement of round lesions (ring lesion) was seen in a few cases before surgery. By histopathology, the lesions showed central geographic necrosis, surrounded by epithelioid granuloma and inflammatory cell bands. A large number of eosinophils and scattered multinucleated giant cells were found, especially at the peripheral of the lesion. Charcot-Leyden crystals were present in all case and parasitic migrans was found in one case. Conclusions: Visceral parasitic migration of liver is a rare liver disease and is easily misdiagnosed as other benign or malignant liver tumors. Combining clinical data, enhanced CT images and pathological examination can improve the preoperative and postoperative diagnosis of the disease.

Anti-proliferative effect of digoxin on breast cancer cells via inducing apoptosis.

OBJECTIVE: Digoxin is a kind of plant-derived cardiac glycoside that is mainly used to treat heart diseases, especially in congestive heart failure or arrhythmia. However, its potentiality presented in anti-tumor remains unexplored. The purpose of this study was designed to investigate the beneficial pharmacological activity of digoxin on breast cancer cell line (MDA-MB-231, MM231). MATERIALS AND METHODS: The methyl thiazolyl tetrazolium (MTT) assay was utilized to detect the proliferation of the breast cancer MM231. The apoptotic cell numbers were determined by the flow cytometry analysis. The expressions of Bcl-2 (B-cell lymphoma-2) and Bax (Bcl2-associated X protein) were detected by Western blot analysis. RESULTS: Digoxin dose-dependently blocked the cell growth of the breast cancer MM231 through MTT assay, whereas the apoptotic numbers were significantly elevated as reflected in acridine orange staining and flow cytometry analysis. In addition, findings from Western blotting method indicated that digoxin intervention showed reduced Bcl-2 expression and elevated Bax level in MM231 cells, characterized by increased Bax/Bcl-2 ratio. CONCLUSIONS: Digoxin plays a potential anti-tumor role in breast cancer in vitro, possibly by inducing mitochondria-dependent apoptosis.

[Clinical application and observation of injectable modified sodium hyaluronate gel filler for facial cosmetic surgery].

Objective: To evaluate the clinical effect of injectable modified sodium hyaluronate gel filler in the treatment of facial profile modification and rejuvenation. Methods: A total of 125 patients who received facial injection of hyaluronate gel from October 2013 to October 2015 were collected. The patients included 62 cases for rhinoplasty, 28 for chin augmentation, 20 for nasolabial fold correction and 15 for lacrimal groove correction. The post-operation results, satisfaction survey and adverse reaction were observed. Results: All the injected positions improved immediately, and the instant average satisfaction score was 9.3±0.7, followed by 8.1±0.7 after 3 months, 6.9±0.8 after 6 month and 5.2±0.8 after 1 year. Thirty cases exhibited swelling, 5 cases bruised, and they all recovered within one week. Conclusions: Sodium hyaluronate is effective and stable in the treatment of facial contour modification and rejuvenation.

Effects of phenol on metabolic activities and transcription profiles of cytochrome P450 enzymes in Chironomus kiinensis larvae.

Phenol, also known as carbolic acid or phenic acid, is a priority pollutant in aquatic ecosystems. The present study has investigated metabolic activities and transcription profiles of cytochrome P450 enzymes in Chironomus kiinensis under phenol stress. Exposure of C. kiinensis larvae to three sublethal doses of phenol (1, 10 and 100 µM) inhibited cytochrome P450 enzyme activity during the 96 h exposure period. The P450 activity measured after the 24 h exposure to phenol stress could be used to assess the level (low or high) of phenol contamination in the environment. To investigate the potential of cytochrome P450 genes as molecular biomarkers to monitor phenol contamination, the cDNA of ten CYP6 genes from the transcriptome of C. kiinensis were identified and sequenced. The open reading frames of the CYP6 genes ranged from 1266 to 1587 bp, encoding deduced polypeptides composed of between 421 and 528 amino acids, with predicted molecular masses from 49.01 to 61.94 kDa and isoelectric points (PI) from 6.01 to 8.89. Among the CYP6 genes, the mRNA expression levels of the CYP6EW3, CYP6EV9, CYP6FV1 and CYP6FV2 genes significantly altered in response to phenol exposure; therefore, these genes could potentially serve as biomarkers in the environment. This study shows that P450 activity combined with one or multiple CYP6 genes could be used to monitor phenol pollution.

Therapeutic effect of inhaled budesonide (Pulmicort® Turbuhaler) on the inflammatory response to one-lung ventilation.

This prospective, double-blind trial was designed to evaluate the effect of inhaled budesonide on lung function and the inflammatory response to one-lung ventilation. One hundred patients scheduled for lobectomy were allocated randomly to pre-operative nebulised budesonide or saline. Bronchoalveolar lavage fluid samples were collected from either the collapsed or the ventilated lung both before one-lung ventilation and 30 min after re-expansion of the lung. The concentrations of serum and bronchoalveolar lavage fluid cytokines were determined. Budesonide treatment, compared with saline, reduced both peak (mean (SD) 3.7 (0.4) vs 2.5 (0.2) kPa) and plateau (mean (SD) 3.1 (0.2) vs 2.2 (0.1) kPa, respectively, p < 0.001 for both) ventilatory pressures. Thirty minutes after re-expansion, lung compliance increased in the budesonide group compared with saline (57.5 (4.1) vs 40.1 (3.5) ml.cmH(2) O(-1), respectively p < 0.001). Budesonide also reduced the concentrations of tumour necrosis factor-α, interleukin-1ß, interleukin-6 and interleukin-8 in bronchoalveolar lavage fluid, but increased interleukin-10 30 min after re-expansion (p < 0.05 for all measures). Pre-operative nebulisation of budesonide may be effective in improving ventilatory mechanics and reducing the inflammatory response to one-lung ventilation during thoracic surgery.

Diterpenoids from Isodon eriocalyx var. laxiflora.

Three new, seco-ent-kaurane diterpenoids, laxiflorins A, B and C, together with four known diterpenoids eriocalyxin B, oriodonin, and maeocrystals A and B, were isolated from the leaves of Isodon eriocalyx var. laxiflora. Their structures were assigned by a combination of one- and two-dimensional NMR techniques and computer modeling calculations. Laxiflorin C displayed weak cytotoxic activity.

Cytotoxic ent-kaurene diterpenoids from three Isodon species.

From Isodon loxothyrsa, one new diterpenoid, loxothyrin A, together with one known diterpenoid, adenolin B, from I.pleiophyllus, three known diterpenoids, coetsoidins A, B and G, and from I. adenoloma, one known diterpenoid, longikaurin F, were isolated. The structure determination of loxothyrin A, and the unambiguous NMR spectral assignments of the known compounds were made by a combination of 1D and 2D NMR techniques and computer modelling calculations. The isolates showed potent cytotoxic activities.

Cytotoxic diterpenoids from Isodon megathyrsus.

A new diterpenoid, megathyrin A, together with three known compounds, rabdocoetsins B, C, and D, were isolated from the leaves of Isodon megathyrsus, and their structures and nmr spectral data were assigned by a combination of one- and two-dimensional nmr techniques. These compounds displayed significant cytotoxic activity.

Epidermal growth factor in the neonatal mouse salivary gland and kidney.

In the adult mouse, epidermal growth factor (EGF) is synthesized in granular convoluted tubule (intralobular) duct cells of the submandibular gland and in distal tubule cells of the kidney. The presence of EGF in developing tissues and maternal milk and the localization of EGF receptors in developing tissues suggest a role for EGF in developmental processes. The primary aims of the present study were to: (1) localize EGF and EGF-binding sites in the kidney and submandibular gland during neonatal development and (2) to determine the effect of exogenously administered EGF on cell proliferation in these two developing organs. In the present study, EGF was localized by immunocytochemistry in granular convoluted tubule cells of the submandibular gland initially on day 21 after birth and in distal tubule cells of the kidney on postnatal day 6. EGF binding in the kidney decreased after birth with some localization to the glomerulus. In submandibular glands of newborn and 10-day-old mice, EGF-binding sites were associated with both acinar and duct cells with peak binding at 10 days postnatally. Submandibular glands from 20-day-old mice demonstrated primarily ductal EGF-binding sites. Exogenously administered EGF induced a mitogenic response in acinar and interlobular duct cells of submandibular glands during the first week after birth. EGF treatment during this period had an inhibitory effect on 3H-thymidine incorporation into cellular compartments in the developing kidney. The identification of EGF-binding sites in the kidney and submandibular gland before the presence of EGF suggests that an EGF-like molecule such as transforming growth factor alpha (TGF-alpha) may be present as a potential ligand in these organs. In order to assess this possibility, developing kidneys and submandibular glands were stained with anti-TGF-alpha. These immunocytochemical studies localized TGF-alpha to the proximal tubule of the kidney and immature acinar cells of the newborn mouse. Our data strongly support an autocrine, juxtacrine or paracrine role for EGF and/or TGF-alpha in the regulation of cell proliferation and cytodifferentiation in the kidney and submandibular gland.

Defective epidermal growth factor gene expression in mice with polycystic kidney disease.

The C57BL/6J-cpk mouse has an inheritable form of polycystic kidney disease similar to the autosomal recessive disorder seen in humans. Between approximately 1 and 3 weeks of age, affected cpk mice develop numerous large cysts in the collecting tubule segment of kidney nephrons. The present study examined the ontogeny of renal and submandibular gland prepro-epidermal growth factor (preproEGF) gene expression in the cpk mouse using Northern blot hybridization and immunohistochemistry. There was a virtual absence of renal preproEGF gene expression in cystic kidneys over the 3-week postnatal period, during which time renal preproEGF mRNA and proEGF/EGF protein normally reach significant levels. PreproEGF mRNA was expressed in salivary glands of cystic mice; however, this mRNA could not be further elevated with testosterone suggesting that there are abnormalities in the regulation of the preproEGF gene in the submandibular gland, as well as in the kidney. Since renal preproEGF expression during the early postnatal period occurs when collecting duct cysts form, it is possible that a deficiency in renal proEGF or EGF contributes to the rapid development of collecting duct cysts and the concomitant renal failure in the C57BL/6J-cpk cystic mouse.

[Clinical investigation of prostate carcinoma in the Third Teaching Hospital of Norman Bethune University of Medical Sciences].

Clinical and statistical investigations were performed on 157 patients with prostate carcinoma in the Third Teaching Hospital, Normal Bethune University experienced between January, 1950 and June, 1986. The number of patients with prostate carcinoma among other hospitalized patients showed a recent gradual increase. The patient's age at the time the disease was first diagnosed was most frequently between 60 and 69 years old with an average age of 63.3 years. Dysuria was the most prominent symptom, followed by frequency, retention and macroscopic hematuria. Duration between initial symptom and diagnosis was one to two years in most patients. The prostatic abnormality could be detected by rectal examination in all patients. Elevation of serum acid phosphatase was found in 24.4%. Such elevation was evident in 52.2% of the patients with metastatic lesions, compared to 14.4% of those without metastasis. Fourteen patients had metastasis to bone (8.9%), 13 to lymph nodes, 2 to lung and one to liver. According to the staging diagnosis, 19 patients (12.1%) had stage A, 78 patients (49.7%) had stage B, 20 patients (12.7%) had stage C and 40 patients (25.5%) had stage D carcinoma. Histological findings in 57 patients indicated adenocarcinomas; 39 cases (68.4%) were poorly differentiated, 12 cases (21.2%) were moderately differentiated and 6 cases (10.5%) were well differentiated. Modality of treatment was total prostatectomy in 2 cases (1.3%), antiandrogen therapy (orchiectomy and/or Stilbestrol) in 122 cases (77.7%), subcapsular prostatectomy in 7 cases (4.5%), symptomatic treatment in 5 cases and no treatment in 23 cases (14.6%).