Objective To investigate the correlations of plasma methoxyadrenaline (MN) and methoxynorepinephrine (NMN) (collectively called MNs) to blood lipid, blood glucose and blood pressure in patients with pheochromocytoma (PCC) and paraganglioma (PGL) (collectively called PPGL). Methods The clinical data were retrospectively analyzed of 64 patients with pathologically confirmed PPGL in Chinese PLA General Hospital during Jan. 2017 to Jun. 2018. According to the tertiles plasma MN before operation, the 64 cases were divided into T1 (n=21), T2 (n=21) and T3 (n=22) group, and then were divided into T’1 (n=21), T’2 (n=21) and T’3 (n=22) group according to the tertiles plasma NMN before operation. The correlations were analyzed of plasma level of MNs to the blood lipid, blood glucose and blood pressure. Results The body mass index (BMI) value decreased significantly with the increase of plasma MNs level in patients with PPGL. With the increase of plasma NMN level, high density lipoprotein cholesterol (HDL-C) and systolic blood pressure (SBP) increased significantly (P<0.05). Pearson correlation analysis showed that the plasma MN was negatively correlated with BMI (r=-0.319, P=0.010), and positively correlated with the tumor diameter (r=0.268, P=0.032). The plasma NMN was negatively correlated with BMI (r=-0.353, P=0.004), and positively correlated with SBP, fasting blood glucose (FBG), total cholesterol (TG), HDL-C, and tumor diameter (r=0.256, 0.019, 0.047, 0.001 and 0.003, respectively. P<0.05). The multivariate linear regression analysis showed that the plasma MN was negatively correlated with BMI (β=0.09, P=0.004) and positively correlated with tumor diameter (β=0.04, P=0.016). Plasma NMN was negatively correlated with BMI (β=0.08, P=0.006), and positively correlated with SBP, FBG, TG, HDL-C, tumor diameter (β=0.01, 0.13, 0.18, 0.50 and 0.06, respectively. P<0.05). Conclusions The plasma MNs level is related to the regulation of blood lipid, blood sugar and blood pressure. Patients with high plasma MNs level before operation should pay more attention to the occurrence of hypertension and high FBG.

Objective: To investigate the effect of hsa_circ_0000392 (circ_0000392) on the radiosensitivity of cervical cancer cells and explore its potential mechanism. Methods: Cervical cancer tissues and adjacent normal tissues of 42 patients with cervical cancer who were confirmed pathologically for the first time in Huaihe Hospital of Henan University from 2016 to 2019 were collected. According to the patients' response to radiotherapy, the cancer tissues were divided into radio-sensitive tissues and radio-resistant tissues. The expressions of circ_0000392, miR-145-5p, and CRKL in radiation-sensitive, radiation-resistant cervical cancer tissues and Hela, SiHa cells were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and western blot. SiRNA circ_0000392, miR-145-5p mimic, miR-145-5p inhibitor, pcDNA 3.1-CRKL and its negative control were transfected into HeLa and Siha cells, respectively. After radiation induction, the survival fraction of cells was detected by clone formation assay, apoptosis was detected by flow cytometry, and the expressions of apoptosis-related proteins Bax and Bcl-2 and ERK pathway protein p-ERK1/2 and ERK1/2 were detected by western blot. The targeting relationship between circ_0000392, miR-145-5p and CRKL was verified by dual luciferase reporter gene assay. The effect of circ_0000392 on radiotherapy sensitivity of cervical cancer in vivo was observed in the tumor formation experiment in nude mice. Results: circ_0000392 and CRKL were upregulated in radiation-resistant tissues and cancer cells of cervical cancer, while miR-145-5p was downregulated. The clone formation numbers of Hela and SiHa cells in si-circ_0000392#1+ 6 Gy group were (78.67±10.97) and (71.00±9.54), respectively, which were lower than those in si-Ctrl+ 6 Gy group [(176.00±22.27) and (158.33±17.56), respectively]. The apoptosis rates were (41.55±3.40)% and (31.41±3.29)%, respectively, which were higher than those in si-Ctrl+ 6 Gy group [(15.91±1.37)% and (13.70±1.89)%, P<0.05]. The protein expression of Bax was higher than that of si-Ctrl+ 6 Gy group, and the protein expressions of Bcl2 was lower than those of si-Ctrl+ 6 Gy group. The clone formation numbers of Hela and SiHa cells in si-circ_0000392#1+ miR-145-5p inhibitor+ 6 Gy group were (171.33±25.01) and (137.00±21.66), higher than those in si-circ_0000392#1+ inhibitor NC+ 6 Gy group [(84.67±17.79) vs (71.00±11.00), P<0.05]. The apoptosis rates were (17.41±2.58) % and (15.96±1.25) %, lower than those of si-circ_0000392 #1+ inhibitor NC+ 6 Gy [(40.29±2.92)% and (30.82±2.34)%, respectively, P<0.05]. The expression of Bax protein was lower than that of si-circ_0000392#1+ inhibitor NC+ 6 Gy group, and the expressions of Bcl2 protein were higher than those of si-circ_0000392#1+ inhibitor NC+ 6 Gy group. Circ_0000392 can target miR-145-5p, and CRKL is the downstream target gene of miR-145-5p. The clone formation numbers of Hela and SiHa cells in miR-145-5p mimic+ 6 Gy group were (74.33±10.02) and (66.00±12.17), respectively, which were lower than those of mimic NC+ 6 Gy group [(197.67±17.21) vs (157.67±11.59), respectively, P<0.05]. The apoptosis rates were (45.58±2.16)% and (32.10±3.55)%, higher than those of mimic NC+ 6 Gy group [(15.85±2.45)% and (13.99±1.69)%, respectively, P<0.05]. The expression of Bax protein was higher than that of the mimic NC+ 6 Gy mimic group, and the expression of Bcl2 protein was lower than that of the mimic NC+ 6 Gy group. The clone formation numbers of Hela and SiHa cells in miR-145-5p mimic+ pcDNA-CRKL+ 6 Gy group were (158.00±15.88) and (122.33±13.65), respectively, which were higher than those of miR-145-5p mimic+ pcDNA+ 6 Gy group [(71.33±8.02) vs (65.67±12.22), P<0.05]. The apoptosis rates were (19.50±3.45)% and (17.04±0.94)%, respectively, which were lower than those of miR-145-5p mimic+ pcDNA+ 6 Gy group [(44.33±2.36)% and (32.05±2.76)%, respectively, P<0.05]. The expression of Bax protein was lower than that of miR-145-5p mimic+ pcDNA group+ 6 Gy group, and the expression of Bcl2 protein was higher than that of miR-145-5p mimic+ pcDNA+ 6 Gy group. Sh-circ_0000392 group had smaller tumor volume and decreased tumor weight (P<0.05). The relative mRNA expression levels of circ_0000392, miR-145-5p and CRKL and the relative protein expression levels of CRKL, Bcl-2 and p-ERK1/2 were decreased, while the relative expression level of Bax protein was increased (P<0.05). Conclusion: Circ_0000392 could enhance the radiosensitivity of cervical cancer cells, and its mechanism may be related to the regulation of CRKL/ERK signaling pathway by targeting miR-145-5p, which provides a new reference for enhancing the radiosensitivity of cervical cancer cells.
Animals ; Mice ; Female ; Humans ; Uterine Cervical Neoplasms/radiotherapy* ; bcl-2-Associated X Protein/genetics* ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2/genetics* ; Apoptosis ; MicroRNAs/genetics* ; Cell Proliferation ; Cell Line, Tumor

Objective: To investigate the effect of hsa_circ_0000392 (circ_0000392) on the radiosensitivity of cervical cancer cells and explore its potential mechanism. Methods: Cervical cancer tissues and adjacent normal tissues of 42 patients with cervical cancer who were confirmed pathologically for the first time in Huaihe Hospital of Henan University from 2016 to 2019 were collected. According to the patients' response to radiotherapy, the cancer tissues were divided into radio-sensitive tissues and radio-resistant tissues. The expressions of circ_0000392, miR-145-5p, and CRKL in radiation-sensitive, radiation-resistant cervical cancer tissues and Hela, SiHa cells were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and western blot. SiRNA circ_0000392, miR-145-5p mimic, miR-145-5p inhibitor, pcDNA 3.1-CRKL and its negative control were transfected into HeLa and Siha cells, respectively. After radiation induction, the survival fraction of cells was detected by clone formation assay, apoptosis was detected by flow cytometry, and the expressions of apoptosis-related proteins Bax and Bcl-2 and ERK pathway protein p-ERK1/2 and ERK1/2 were detected by western blot. The targeting relationship between circ_0000392, miR-145-5p and CRKL was verified by dual luciferase reporter gene assay. The effect of circ_0000392 on radiotherapy sensitivity of cervical cancer in vivo was observed in the tumor formation experiment in nude mice. Results: circ_0000392 and CRKL were upregulated in radiation-resistant tissues and cancer cells of cervical cancer, while miR-145-5p was downregulated. The clone formation numbers of Hela and SiHa cells in si-circ_0000392#1+ 6 Gy group were (78.67±10.97) and (71.00±9.54), respectively, which were lower than those in si-Ctrl+ 6 Gy group [(176.00±22.27) and (158.33±17.56), respectively]. The apoptosis rates were (41.55±3.40)% and (31.41±3.29)%, respectively, which were higher than those in si-Ctrl+ 6 Gy group [(15.91±1.37)% and (13.70±1.89)%, P<0.05]. The protein expression of Bax was higher than that of si-Ctrl+ 6 Gy group, and the protein expressions of Bcl2 was lower than those of si-Ctrl+ 6 Gy group. The clone formation numbers of Hela and SiHa cells in si-circ_0000392#1+ miR-145-5p inhibitor+ 6 Gy group were (171.33±25.01) and (137.00±21.66), higher than those in si-circ_0000392#1+ inhibitor NC+ 6 Gy group [(84.67±17.79) vs (71.00±11.00), P<0.05]. The apoptosis rates were (17.41±2.58) % and (15.96±1.25) %, lower than those of si-circ_0000392 #1+ inhibitor NC+ 6 Gy [(40.29±2.92)% and (30.82±2.34)%, respectively, P<0.05]. The expression of Bax protein was lower than that of si-circ_0000392#1+ inhibitor NC+ 6 Gy group, and the expressions of Bcl2 protein were higher than those of si-circ_0000392#1+ inhibitor NC+ 6 Gy group. Circ_0000392 can target miR-145-5p, and CRKL is the downstream target gene of miR-145-5p. The clone formation numbers of Hela and SiHa cells in miR-145-5p mimic+ 6 Gy group were (74.33±10.02) and (66.00±12.17), respectively, which were lower than those of mimic NC+ 6 Gy group [(197.67±17.21) vs (157.67±11.59), respectively, P<0.05]. The apoptosis rates were (45.58±2.16)% and (32.10±3.55)%, higher than those of mimic NC+ 6 Gy group [(15.85±2.45)% and (13.99±1.69)%, respectively, P<0.05]. The expression of Bax protein was higher than that of the mimic NC+ 6 Gy mimic group, and the expression of Bcl2 protein was lower than that of the mimic NC+ 6 Gy group. The clone formation numbers of Hela and SiHa cells in miR-145-5p mimic+ pcDNA-CRKL+ 6 Gy group were (158.00±15.88) and (122.33±13.65), respectively, which were higher than those of miR-145-5p mimic+ pcDNA+ 6 Gy group [(71.33±8.02) vs (65.67±12.22), P<0.05]. The apoptosis rates were (19.50±3.45)% and (17.04±0.94)%, respectively, which were lower than those of miR-145-5p mimic+ pcDNA+ 6 Gy group [(44.33±2.36)% and (32.05±2.76)%, respectively, P<0.05]. The expression of Bax protein was lower than that of miR-145-5p mimic+ pcDNA group+ 6 Gy group, and the expression of Bcl2 protein was higher than that of miR-145-5p mimic+ pcDNA+ 6 Gy group. Sh-circ_0000392 group had smaller tumor volume and decreased tumor weight (P<0.05). The relative mRNA expression levels of circ_0000392, miR-145-5p and CRKL and the relative protein expression levels of CRKL, Bcl-2 and p-ERK1/2 were decreased, while the relative expression level of Bax protein was increased (P<0.05). Conclusion: Circ_0000392 could enhance the radiosensitivity of cervical cancer cells, and its mechanism may be related to the regulation of CRKL/ERK signaling pathway by targeting miR-145-5p, which provides a new reference for enhancing the radiosensitivity of cervical cancer cells.
Animals ; Mice ; Female ; Humans ; Uterine Cervical Neoplasms/radiotherapy* ; bcl-2-Associated X Protein/genetics* ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2/genetics* ; Apoptosis ; MicroRNAs/genetics* ; Cell Proliferation ; Cell Line, Tumor

Objective:To observe the efficacy of modified Simotang in treatment of adult functional constipation (Qi-stagnancy constipation), and investigate its effects on serum levels of intestinal neurotransmitter nitric oxide synthase (nNOS), nitric oxide (NO), and vasoactive intestinal peptide (VIP), as well as superoxide dismutase (SOD), malonaldehyde (MDA), and glutathione (GSH) levels. Method:One hundred and ten patients with functional constipation were selected and randomly divided into control group (55 cases) and treatment group (55 cases) by referring to random number table. The patients in control group were given with routine therapy, Domperidone tablet (1 tablet/time, tid), and Phenolphthalein tablets (100 mg/time, bid). The patients in treatment group were treated with modified Simotang, 1 dose/day. Both groups were treated for 4 weeks. Then the scores of main clinical symptoms of functional constipation, scores of Qi-stagnancy constipation and clinical efficacy were compared between two groups. Constipation recurrence rate was compared between two groups after stopping medicine. Serum levels of intestinal neurotransmitters nNOS, NO and VIP as well as SOD, MDA, GSH levels were detected in both groups. Result:After treatment, scores for main clinical symptoms (difficult defecation, abdominal distension, defecation time, number of defecation times) and Bristol scores in treatment group were obviously lower than those in control group (P<0.01). Scores for symptoms of Qi-stagnancy constipation (ungratifying defecation, abdominal distension,bowel ringing, frequent flatus, chest and flank tightness) in treatment group were obviously lower than those in control group after treatment (P<0.01). Total clinical efficacy in treatment group 98.04% was superior to that in control group 84.62% ( P<0.05). Constipation recurrence rate was 3.92% and 8.16% in treatment group after 4 and 8 weeks of medication stopping, obviously lower than those in control group 18.18% and 27.78% (P<0.05). After treatment, serum levels of intestinal neurotransmitters nNOS, NO, VIP in treatment group were remarkably lower than those in control group (P<0.01). After treatment, serum levels of SOD and GSH in treatment group were higher while MDA level was lower than those in control group (P<0.01). Conclusion:Based on routine therapy, modified Simotang in treatment of adult functional constipation (Qi-stagnancy constipation) can improve clinical symptoms and syndrome symptoms, increase the clinical efficacy, decrease recurrence rate, and regulate levels of intestinal neurotransmitters nNOS,NO and VIP as well as SOD, MDA, GSH levels.

OBJECTIVERecent studies have found that the variation of G894T on the region of T786C and 7th exon promoted by endothelial nitric oxide synthase (eNOS) gene is associated with cardiovascular disease. This research explored possible correlations between eNOS gene polymorphisms and orthostatic intolerance (OI) in children through linkage disequilibrium analysis between eNOS genes T786C and G894T and OI.

METHODSPCR, Macrorestriction Map and other molecular biotechnology were used to determine the genotypes of eNOS/T786C and G894T in 60 OI probands and their parents. Correlation analysis and transmission disequilibrium test (TDT) between T786C, G894T and OI were performed.

RESULTSThere was linkage disequilibrium of eNOS/T786C and G894T gene polymorphisms in the occurrence of childhood OI (P<0.05).

CONCLUSIONSeNOS genes T786C and G894T may be associated with the pathogenesis of OI.

Adult ; Child ; Female ; Humans ; Linkage Disequilibrium ; Male ; Nitric Oxide Synthase Type III ; genetics ; Orthostatic Intolerance ; genetics ; Polymorphism, Genetic

Autoimmune diseases are a kind of chronic diseases with unclear etiology, which has the characteristics of repetition and difficulty to cure completely. Aerobic exercise, as an effective intervention method for chronic diseases, has also received extensive attention in the field of the prevention and treatment of autoimmune diseases. In this paper, the effects of aerobic exercise on immune system and autoimmune diseases in recent years are reviewed, and the related mechanisms are discussed. It is pointed out that aerobic exercise can improve the homeostasis of immune environment by affecting the number and function of immune cells, inhibit the systemic inflammatory response of the body, and then delay the occurrence and development of autoimmune diseases.
Autoimmune Diseases ; prevention & control ; Exercise ; Homeostasis ; Humans ; Immune System

OBJECTIVETo observe the absorption and concentration of berberine hydrochloride (BH) in gastric, entric, colonic tissue after intragastric administration of BH-containing carboxymethyl konjac glucomannan pellets for evaluating colon-specific drug delivery characteristics of the pellets.

METHODBH-containing carboxymethyl konjac glucomannan pellets (pellets group) and BH-containing carboxymethyl cellulose suspension (control group) were intragastric administrated to rats at the dose of 50 mg x kg(-1), respectively. A high performance liquid chromatography method determinated BH concentration in rat plasma and tissue. Drug delivery index (DDI) was calculated.

RESULTThe range of BH in plasma and tissue in rats were 0.025 2-2.52 mg x L(-1) (r = 0.999 2) and 0.126-25.22 mg x L(-1) (R > 0.99),respectively. The detection of BH in plasma and tissue were 10 microg x L(-1) and 8 microg x L(-1), respectively. The area under the curve (AUC(0 --> infinity)) in the plasma samples of pellets group was 0.477 times that of the control group; in the gastric, entric, colonic tissue, the AUC(0 --> infinity) of pellets group was as much as 0.187, 0.228, 2.00 times that of the control group, respectively. The DDI of the pellets was 0.392 4, 0.478 6, 4.193 in the gastric, entric, colonic tissue of the rat, respectively.

CONCLUSIONCarboxymethyl konjac glucomannan pellets may be a useful carrier of BH for colon-specific delivery.

Absorption ; Animals ; Berberine ; metabolism ; Calibration ; Chromatography, High Pressure Liquid ; Drug Implants ; Female ; Intestines ; metabolism ; Male ; Mannans ; administration & dosage ; pharmacokinetics ; Organ Specificity ; Rats ; Sensitivity and Specificity

OBJECTIVETo study release mechanism of berberine hydrochloride (BH) from carboxymethyl konjac glucomannan pellets for colonic delivery.

METHODThe pellets were prepared by ionotropic gelation technique. The effects of the kinds of enzyme and enzyme concentration of dissolution media on the release of BH and the erosion properties of the pellets were studied.

RESULTCompared with the dissolution media without enzymes, the release of BH and the erosion of the pellets were increased obviously in the media with rat cecal and colonic content or beta-mannase, the degradation of the carrier material of pellets by enzymes was the main factor which result in the erosion of the pellets. With the increased of beta-mannase concentration, the release of BH and the erosion of the pellets increased, the amount relationships of the release of BH and the erosion of the pellets were approximately 1:1. The release of BH exhibit Peppas equation, the n value was more than 1.

CONCLUSIONThe release mechanism of BH from the pellets was enzymatic erosion-controlled, which indicates the potential of the pellets to serve as a colon-specific drug delivery system.

Animals ; Berberine ; administration & dosage ; pharmacokinetics ; Biological Transport ; drug effects ; Colon ; metabolism ; Drug Delivery Systems ; methods ; Mannans ; chemistry ; Rats ; Rats, Sprague-Dawley ; beta-Mannosidase ; pharmacology

Objective To investigate the effects of subcutaneous transfection of human beta-nerve growth factor (Ad-hNGFβ) gene on the expression of substance P (SP) in the dorsal root ganglion in a rat model of diabetic neuropathic pain (DNP).Methods Male SD rats weighing 180-220 g were used in this study.Ten rats were randomly collected as normal control without DNP (group C).DNP model was made by intraperitoneal injection of streptozocin (STZ) 75 mg/kg.Seventy-five rats with DNP were randomly divided into 3 groups ( n =25 each):DNP group; Ad-hNGF group and Ad-LacZ group.Groups Ad-NGF and Ad-LacZ received subcutaneous transfection of 1.12 × 1010 PFU Ad-hNGFβ 10 μl and 1.12 × 1010 PFU Ad-LacZ 10 μl respectively after pain threshold was measured on 21d after STZ injection.The mechanical and thermal pain threshold was measured before STZ injection (baseline) and at 21,35 and 49 d after STZ injection.The expression of SP in the dorsal root ganglion was determined after the measurement of pain threshold on 49 d after STZ injection.Results Compared with group C,the mechanical and thermal pain threshold was significantly decreased at each time point after STZ injection in groups DNP,Ad-NGF and Ad-LacZ,and the expression of SP in the dorsal root ganglion was signilicantly downregulated in groups DNP and Ad-LacZ (P ＜ 0.05).Compared with group DNP,the thermal pain threshold was significantly increased on 49 d afar STZ injection,and the expression of SP in the dorsal root ganglion was significantly up-regulated in group Ad-NGF ( P ＜ 0.01 ),and no significant change was found in the mechanical and thermal pain threshold and the expression of SP in the dorsal root ganglion at each time point in group Ad-LacZ ( P ＞ 0.05).Conclusion Subcutaneous transfection of Ad-hNGFβ can attenuate DNP to some extent through upregulation of the expression of SP in rat dorsal root ganglion.

OBJECTIVETo prepare sinomenine hydrochloride delayed-onset sustained-release tablets.

METHODThe tablets containing sinomenine hydrochloride were prepared by dry-compression coating technique with the ratio of HPMC in core tablet and the ratio of HPMC in coating film as the influence factors and the lag-time and release rate as the evaluation parameters. Experiments were done on the central composite design, the data were simulated by using multi-linear equation and second-order polynomial equation. The possibly optimal formulation was predicted by response surface method. The dissolution date (lag-time and release rate) of the tablets prepared under the optimum condition were compared with the predicted. The drug released mechanism of the tablet were studied by Model-fitted of drug released within 6-15 h with zero-order, Higuchi and Peppas equation, respectively.

RESULTThe lag-time and release rate were simulated using second-order polynomial equation, regression coefficients of the two parameters were 0.9901 and 0.9876, respectively. Bias between the observed and predicted values of lag-time and release rate were -3.15% and -0.34%, respectively. The lag-time of the tablet prepared under the optimum condition in vitro was about 6 h, then drug released from the tablet within 6-15 h was found to conform to zero-order kinetics and was controlled by bulk erosion mechanism.

CONCLUSIONSinomenine hydrochloride delayed-onset sustained-release tablets release drug slowly after lag time. The models developed in this study are proved to be highly predictable.

Chemistry, Pharmaceutical ; Delayed-Action Preparations ; administration & dosage ; Drug Delivery Systems ; methods ; Excipients ; administration & dosage ; Morphinans ; administration & dosage ; Pharmaceutical Preparations ; administration & dosage ; Tablets ; administration & dosage ; Technology, Pharmaceutical