Objective To study the changes of cytokine activation in patients with chronic congestive haert failure(CHF) as indicated by plasma levels of C-reactive protein(CRP)and its relation to adrenomedullin(ADM) and endthelin-1(ET-1).Methods The plasma levels of CRP,ADM and ET-1 of 60 decompensated haert failure(DHF) patients group and 30 essential hypertension patients with compensated cardiac function(CCF) group were tested. CRP was determined by immunoturbidometry,ET-1 and ADM were determined by radioimmunoassay.Results The plasma levels of ET-1 ,ADM and CRP of decompensated heart failure group were significantly higher than the compensated cardiac function group;In the decompensated heart failure group,the plasma levels of CRP and ADM were elevated according to worsening of heart failure(NYHA classification) ;In the decompensated heart failure group,the plasma levels of CRP were positively correlated with ADM(r=0.57).Conclusion There is extensive activation of cytokines,ADM and ET-1 in patients with chronic congestive heart failure.Cytokine activation might play a role in the synthesis of ADM,thus contribute to the hum oral regulation of heart failure,but may not be re- sponsible for the activation of ET-1.

AIMTo prepare salcatonin dry powder inhalations (sCT-DPIs) A (mixture of mannitol and L-leucine) and B (mixture of manntiol and lactose) by spray-drying and then to study their main pharmaceutical properties.

METHODSDumping rate of sCT-DPIs capsules and deposited fraction of sCT at effective part were determined according to Chinese Pharmacopiea 2000. Particle morphology under different relative humidity (RH) was observed by scanning electronics microphotograph, particle size and its distribution were determined by Malvern Mastersizer and the transition of morphorous state for carriers before and after spray-drying was investigated by differential thermal analysis (DTA) and X-ray powder diffraction (XRPD).

RESULTSDumping rates of sCT-DPIs A and B capsules were both above 10% and deposited fraction of sCT at effective part was above 90% for both A and B, which were all in agreement with the standard of Chinese Pharmacopiea 2000. Powder particle of sCT-DPIs A was round and existed one by one after keeping one month under RH 0, 23% and 52%, but aggregation can be observed under RH 75%; many particles which were also round agglomerated in sCT-DPIs B even under zero RH; mean particle size of sCT-DPIs A was 1.67 microns, which was much smaller than that of sCT-DPIs B; In sCT-DPIs A particle with empty core which was lighter than the same size particle with concreted core was observed. It was shown by DTA that melted heat of L-leucine in sCT-DPIs composed of mannitol and L-leucine lowered much more than that of L-leucine exisited alone after spray-drying. It was confirmed by XRPD that diffraction intensity of carriers in sCT-DPIs decreased more than that of carriers before spray-drying.

CONCLUSIONRound particle can be made when mannitol was added to carriers and ultra low density carriers can be formed when L-leucine was added. It was suggested by SEM that DPIs should be kept under certain RH. Particle size and distribution of sCT-DPIs all accorded with demand of DPIs. Complex spray-drying carriers formed amorphous state easier than single carrier.

Administration, Inhalation ; Anti-Asthmatic Agents ; administration & dosage ; Calcitonin ; administration & dosage ; Differential Thermal Analysis ; Leucine ; Mannitol ; Microscopy, Electron, Scanning ; Particle Size ; Powders ; Technology, Pharmaceutical ; methods

Objective To investigate the effect of caveolin-1 on rabbit carotid artery anastomotic stenosis and its relationship with TNF-α.Methods 40 New Zealand white rabbits were randomly divided into normal group,surgical group,empty vector group and caveolin-1 transfecting group.Carotid artery end-to-end anastomosis was done in the rabbits except these in normal group.Five specimens were randomly taken on day 7 after surgery for Westem blot to detect the expression of caveolin-1 and TNF-α; The rest specimens were taken for HE staining at fourth week.The ratio of intima/media area were measured by Image-Pro Plus 6.0 software in order to observe the proliferation of intima.Results Compared with normal group,in surgical group intimal proliferation was significant,the intima/media ratio was significantly higher (P ＜ 0.05) ; Compared with surgical group,in caveolin-1 transfected group neointimal proliferation was not obvious,the intima/media ratio decreased (P ＜ 0.05).Western blot showed that:compared with the surgical group,caveolin-1 expression in transfected group was significantly higher (P ＜ 0.05) ; compared with normal group,the TNF-α expression in surgical group increased (t:41.28,P ＜ 0.05) ; Compared with surgical group,TNF-α expression in transfected group decreased significantly (t:36.37,P ＜ 0.05).Conclusions Caveolin-1 inhibits vascular anastomotic stenosis,possibly by down-regulating TNF-α expression.

Objective To investigate the effect of Caveolin-1 on extracellular regulated protein kinases of rabbit carotid artery anastomotic restenosis.Methods 40 New Zealand white rabbits were randomly divided into normal control group,carotid artery end to end anastomosis surgical group,empty vector transfection on the site of anastomosis group and Caveolin-1 transfected group.Left carotid artery endto-end anastomosis was performed,and the mixture of Caveolin-1 plasmid and liposome lipofectin 2000 (transfected group) or empty plasmid and lipofectin 2000 mixture (empty vector group) were transfected on anastomosis.Specimens were taken at 7 d after surgery for Western blot and RT-PCR to detect the expression of protein and mRNA.Specimens were taken for HE staining at 28 d to observe the proliferation of intima,and measured the ratio of intima/media area by Image-Pro Plus 6.0 software.Results Compared with surgical group,the ratio of intima/media area in Caveolin-1 transfected group decreased by about 50％.Compared with surgical group and empty vector group,the Caveolin-1 mRNA expression and protein activity significantly increased (t =36.59,P ＜ 0.01) ; the ERK1/2 mRNA expression and protein activity significantly decreased on rabbit carotid artery anastomotic site in Caveolin-1 transfected group (t =32.64and 7.27,P ＜ 0.01).Conclusions Caveolin-1 inhibits anastomotic restenosis possibly by regulating the activation of ERK.

AIM: To investigate the effects of sodium ferulate on cholesterol and triglyceride metabolism in atherosclerosis with hyperlipidemia.METHODS: The rabbit model of atherosclerosis was produced by feeding high lipid forages.RAW264.7 foam cell and HepG2 injured cell models were established by incubation with oxidized low density lipoprotein(ox-LDL).The atherosclerotic plaque area was measured,and serum lipids were detected.The cellular lipid accumulation was examined by oil red O staining.The cellular contents of total cholesterol and cholesterol ester were quantified by high performance liquid chromatography.The hepatic lipase(HL) mRNA expression was determined by RT-PCR.RESULTS:(1) Compared with hyperlipid group,the aorta atherosclerosis plaque area and the serum triglyceride level were significantly decreased in sodium ferulate-treated rabbits,but the serum cholesterol level showed little change.(2) Compared with ox-LDL group,the HL mRNA expression in HepG2 cells was enhanced significantly in sodium ferulate-treated group,but the cellular contents of total cholesterol and cholesterol ester in RAW264.7 foam cells showed little change.CONCLUSION: Sodium ferulate inhibits the formation of atherosclerotic plaque in high-cholesterol-fed rabbits aorta.This antiatherosclerotic function may reduce serum triglyceride level through enhancing the expression of HL mRNA without influencing serum cholesterol level and foam cell formation.

There is a high prevalence of thalassemia in the South of China. To explore the genotype of alpha-thalassemia as well as the distribution of alpha globin gene mutation in the South of China, 356 patients with heterozygote alpha(+) thalassemia, heterozygote alpha(0) or homozygote alpha(+) thalassemia and 78 patients with HbH were analyzed. The gene diagnosis methods including Gap-PCR, nested-PCR, PCR-RE, PCR-SSCP, 4P-ASPCR and DNA sequence analysis were used. The results showed that among 356 patients, 295 patients with --SEA/alphaalpha (82.87%), 1 patient with alphaalpha/alpha-alpha(3.7) (0.28%), 3 patients with alphaalpha/alpha-alpha(4.2) (0.84%), 3 patients with alphaalpha/alpha(CS)alpha (0.84%), 1 patient with alphaalpha/alphaalpha(QS) (0.28%) and 2 patients with alphaalpha/alpha(Westmead) alpha (0.56%) were found. The homozygote with -alpha(4.2) or -alpha(3.7) was not found. In 78 patients with HbH, 29 patients with --SEA/alphaalpha(-3.7) (37.2%), 20 patients with --SEA/alphaalpha(-4.2) (25.6%), 19 patients with --SEA/alphaalpha(CS) (24.3%), 2 patients with --SEA/alphaalpha(QS) (2.6%) were detected, and other remaiming 8 patients were needed to be defined. Among the non-defined 8 patients, the synonymous mutation with C-->G transversion (GCC-GCG) at codon 65 in the exon 2 of alpha 2-globin gene was detected in 2 unrelated HbH patients came from Guangxi province. Whether it correlated with the phenotype of HbH disease or it is only a single nucleotide polymorphism site (SNPs), should be confirmed in the future. In addition, a set of gene diagnosis methods based on PCR to screen deletion and non-deletion genotypes of alpha-thalassemia in Chinese was improved. A new method, 4P-ASPCR, to detect Hb CS and Hb QS was also developed. The method was verified to be more accurate, time-saving and economic. In conclusion, the genotypes of alpha-thalassemia in Chinese are very complicated, the genotypes of alpha-thalassemia in Chinese need to be further studied, the results of this research probably have practical significance for the gene diagnosis or antenatal diagnosis of alpha-thalassemia in the South of China.
Base Sequence ; China ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Gene Deletion ; Gene Frequency ; Genotype ; Globins ; genetics ; Hemoglobin H ; genetics ; Hemoglobins ; genetics ; Hemoglobins, Abnormal ; genetics ; Humans ; Molecular Sequence Data ; Mutation ; Polymorphism, Single-Stranded Conformational ; alpha-Thalassemia ; genetics ; pathology

Objective: To investigate the clinical efficacy of dorsal mosaic surgery with penis free flap for the treatment of anterior urethral stricture after TURP. Methods: We analyzed the clinical data of the patients with anterior urethral stricture after TURP from January 2010 to December 2017 in Yan′an hospital affiliated to Kunming medical university retrospectively. The patients' age ranged from 58 to 75 years, with an average of 64.3 years. The time from TURP to the diagnosis of anterior urethral stricture was 1-12 month, with an average of 3.5 months. 5 cases were urethral stricture at penis segment, 11 cases were urethral stricture at the junction of penis and scrotum, and the length of the narrow urethra was 2-5 cm, with an average of 3.4cm. The average maximum uroflowmetry in preoperative was (5.3±2.7) ml/s.11 cases were treated with regular urethral dilatation and the treatment durable time was more than 6 months, 5 cases were treated with intraurethral incision combined with urethral dilatation(1 or 2 times). 16 cases were not effective after receiving the above treatment, so that all cases were treated with dorsal mosaic surgery with penis free flap. Subarachnoid anesthesia combined with epidural anesthesia, the patient took the supine position.The distal end of urethral stricture was defined by urethral dilator. Incision from the ventral side of the urethra. The length of the incision was extended 0.5 cm based on the length of urethral stricture in urethral angiography.Anatomize the left and right sides of the urethral stricture and longitudinally incision the ventral side of the urethral cavernous body. The length of the incision was extended 0.5 cm to the normal urethral mucosa. The traction line retracts the ventral urethral edge along both sides.The corresponding medial line of the dorsal urethra was incised to the tunica albuginea, and the urethral edge of the dorsal side was separated from the tunica albuginea to form an elliptical region on the tunica albuginea.According to the size of the ellipse, the full thickness of the penis flap was taken, and the size of the flap was beyond the edge of the elliptical area about 0.3 cm.The free flap was covered with the 6-0 absorbable suture on the elliptical area (the skin surface was on the inner side of the urethra), the edge of the free flap was intermittently sutured with the urethral edge of the dorsal side, Multi-needle intermittent sutured flap surface on the corpus cavernosum bed.Using a silicone catheter as the stent tube of the new urethra, the 6-0 absorbable suture closes the ventral side of the incisional urethral sponge.The multi-layered meat film was sutured to prevent leakage of urine, and the fascia and skin were sutured layer by layer. The 5-0 absorbable thread sutures the wound after the foreskin was taken. Results: The operations were successfully completed. The operation time was 90-120 min, with an average of 102.3 min. The intraoperative blood loss was 10-30 ml. The symptoms of dysuria were relieved in all patients after removal of the catheter at 3 weeks postoperative. 4 weeks after surgery, no signs of urethral stricture were observed in urethrography. And the maximum flow rate was >15 ml/s in 13 cases, while 3 cases was 10-15 ml/s. The mean maximum flow rate in postoperative was [(20.4±7.3) ml/s], which was significantly higher than that in preoperative (t=7.7602, P<0.05). B-ultrasound showed 13 cases without residual urine and 3 cases of residual urine volume <30 ml. All patients had no serious complications such as urinary fistula, urethral diverticulum and extravasation of urine. After 1 year of follow-up, 1 patient was lost to follow up, and none of the remaining15 cases had urethral obstruction due to re-stricture. Conclusion Dorsal mosaic surgery with penis free flap could be an effective method and had no obvious complications for the treatment of anterior urethral stricture after TURP.

Objective: To investigate the relation of Epstein-Barr virus (EBV)-induced immune escape to regulatory T cells(Treg) in nasopharyngeal carcinoma(NPC). Methods: We recruited 83 patients who were pathologically diagnozed as nasopharyngeal carcinoma and treated in Nanfang Hospital from January to September 2016. CD4⁺ CD25⁺ FOXP3⁺ Treg in peripheral blood was examined by flow cytometry and the level of EBV DNA in the blood was detected by quantitative real-time polymerase chain reaction(qRT-PCR)in 83 patients and 51 healthy adults as control. The expression of FOXP3⁺ Treg and EBER in NPC in 32 of 83 patients and in nasopharyngitis in 16 control patients were tested by immunohistochemistry(IHC) and in-situ hybridization(ISH). Results: The ratio of Treg in blood in patients with NPC(5.29±1.45)% was higher than that in control group (4.78±1.19) %, with a statistically significant difference(P=0.035). The ratio of Treg in blood in patients with positive EBV was higher than that in patients with negative EBV(P=0.013), and there was a positive correlation between EBV DNA copies and Treg ratio(r=0.335, P=0.002). The density of FOXP3⁺ Treg in NPC was higher than that in nasopharyngitis(P=0.001). Positive correlation was found between the density of FOXP3⁺ Treg and the stain intensity of EBER(r=0.496, P=0.004). Conclusion The ratios of Treg in blood and tissue in NPC patients with positive EBV are significantly increased, indicating a role of Treg in EBV-induced immune escape in NPC.

Objective To evaluate the efficiency of a recombinase-aided amplification (RAA) assay for the detection of Schistosoma japonicum infections in Oncomelania hupensis snails. Methods A group test was employed. Fifty Oncomelania snails were collected as a detection sample. The detection samples without infected snails were designated as negative specimens, while the detection samples that contained different numbers of infected snails were designated as positive specimens. A total of 10 negative specimens, 10 positive specimens containing 1 infected snail, 20 positive specimens containing 2 infected snails and 10 positive specimens containing 3 infected snails were assigned. Following random grouping, 40 specimens were subject to the florescent RAA assay using a blind method. The miradium shedding method served as a gold standard, and the sensitivity, specificity, Youden’s index and coincidence rate of the florescent RAA assay were estimated. In addition, 20 samples consisted of 5 negative specimens and 15 positive specimens with 1, 2 and 3 infected snails respectively were grouped randomly. The same specimens were detected using the crushing method and fluorescent RAA assay with the blind method in a paired-design manner. Then, the test results were compared and analyzed. Results Florescent RAA assay detected 29 positives in the 30 specimens containing different numbers of infected snails, with a sensitivity of 96.67%, and 8 negatives in the 10 detection specimens without infected snails, with a specificity of 80.00%, showing a Youden’s index of 0.77. The coincidence rate was 100% among 10 repeated assays for a detection specimen. In addition, there was no significant difference in the detection of infected snails between the florescent RAA assay and the crushing method (χ² = 0, P > 0.05), and the actual coincidence rates of the florescent RAA assay and crushing method were 95.00% (19/20) and 90.00% (18/20) with the real results, respectively. Conclusion Fluorescent RAA assay has a favorable efficiency for the detection of S. japonicum infections in Oncomelania snails, which shows a potential in screening of S. japonicum-infected Oncomelania snails.

Objective: To analyze the clinical epidemiological characteristics including composition of pathogens , clinical characteristics, and disease prognosis acute bacterial meningitis (ABM) in Chinese children. Methods: A retrospective analysis was performed on the clinical and laboratory data of 1 610 children <15 years of age with ABM in 33 tertiary hospitals in China from January 2019 to December 2020. Patients were divided into different groups according to age,<28 days group, 28 days to <3 months group, 3 months to <1 year group, 1-<5 years of age group, 5-<15 years of age group; etiology confirmed group and clinically diagnosed group according to etiology diagnosis. Non-numeric variables were analyzed with the Chi-square test or Fisher's exact test, while non-normal distrituction numeric variables were compared with nonparametric test. Results: Among 1 610 children with ABM, 955 were male and 650 were female (5 cases were not provided with gender information), and the age of onset was 1.5 (0.5, 5.5) months. There were 588 cases age from <28 days, 462 cases age from 28 days to <3 months, 302 cases age from 3 months to <1 year of age group, 156 cases in the 1-<5 years of age and 101 cases in the 5-<15 years of age. The detection rates were 38.8% (95/245) and 31.5% (70/222) of Escherichia coli and 27.8% (68/245) and 35.1% (78/222) of Streptococcus agalactiae in infants younger than 28 days of age and 28 days to 3 months of age; the detection rates of Streptococcus pneumonia, Escherichia coli, and Streptococcus agalactiae were 34.3% (61/178), 14.0% (25/178) and 13.5% (24/178) in the 3 months of age to <1 year of age group; the dominant pathogens were Streptococcus pneumoniae and the detection rate were 67.9% (74/109) and 44.4% (16/36) in the 1-<5 years of age and 5-<15 years of age . There were 9.7% (19/195) strains of Escherichia coli producing ultra-broad-spectrum β-lactamases. The positive rates of cerebrospinal fluid (CSF) culture and blood culture were 32.2% (515/1 598) and 25.0% (400/1 598), while 38.2% (126/330)and 25.3% (21/83) in CSF metagenomics next generation sequencing and Streptococcus pneumoniae antigen detection. There were 4.3% (32/790) cases of which CSF white blood cell counts were normal in etiology confirmed group. Among 1 610 children with ABM, main intracranial imaging complications were subdural effusion and (or) empyema in 349 cases (21.7%), hydrocephalus in 233 cases (14.5%), brain abscess in 178 cases (11.1%), and other cerebrovascular diseases, including encephalomalacia, cerebral infarction, and encephalatrophy, in 174 cases (10.8%). Among the 166 cases (10.3%) with unfavorable outcome, 32 cases (2.0%) died among whom 24 cases died before 1 year of age, and 37 cases (2.3%) had recurrence among whom 25 cases had recurrence within 3 weeks. The incidences of subdural effusion and (or) empyema, brain abscess and ependymitis in the etiology confirmed group were significantly higher than those in the clinically diagnosed group (26.2% (207/790) vs. 17.3% (142/820), 13.0% (103/790) vs. 9.1% (75/820), 4.6% (36/790) vs. 2.7% (22/820), χ²=18.71, 6.20, 4.07, all P<0.05), but there was no significant difference in the unfavorable outcomes, mortility, and recurrence between these 2 groups (all P>0.05). Conclusions: The onset age of ABM in children is usually within 1 year of age, especially <3 months. The common pathogens in infants <3 months of age are Escherichia coli and Streptococcus agalactiae, and the dominant pathogen in infant ≥3 months is Streptococcus pneumoniae. Subdural effusion and (or) empyema and hydrocephalus are common complications. ABM should not be excluded even if CSF white blood cell counts is within normal range. Standardized bacteriological examination should be paid more attention to increase the pathogenic detection rate. Non-culture CSF detection methods may facilitate the pathogenic diagnosis.
Adolescent ; Brain Abscess ; Child ; Child, Preschool ; Escherichia coli ; Female ; Humans ; Hydrocephalus ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial/epidemiology* ; Retrospective Studies ; Streptococcus agalactiae ; Streptococcus pneumoniae ; Subdural Effusion ; beta-Lactamases