AIM:To investigate the effects of down-regulation of protein kinase C (PKC) on the activity of storeoperated Ca2 + channels (SOC) and the proliferation of airway smooth muscle cells (ASMCs). METHODS:Rat bronchial smooth muscle cells were isolated and cultured. Fluo-3 /AM fluorescence was measured by laser confocal microscope to assessing intracellular Ca2 +. Downregulation of PKC activity was achieved by incubation of ASMCs with PKC activator phorbol-12-myristate-13-acetate (PMA,10 ?mol/L) or phorbol 12,13 -dibutyrate (PDBu,1 ?mol/L) for 24 h. The proliferation of ASMCs was assayed by calculating the reduction rates of Alamar blue. RESULTS:Down-regulation of PKC activity by longterm exposure of PMA or PDBu inhibited the proliferation of ASMCs,the similar results were obtained by using PKC inhibitor chelerythrine. Both downregulation of PKC activity and inhibition of PKC activity by chelerythrine reduced Ca2 + entry through SOC channels. Low concentration of PMA (0. 1 ?mol/L) promoted the proliferation of ASMCs,and this effect was inhibited by SOC blocker SKF-96365. CONCLUSION:Inhibition or down -regu-lation of PKC activity results in the inhibition of SOC channels,suggesting that PKC is involved in the activation of these channels. Ca2 + entry through SOC channels might contribute to PKC-promoted proliferation of ASMCs.

This survey included 59 secondary researches of traditional pharmaceutical patent literatures. The analysis was made on statistical indicator of the amount of annual papers, authors, institutions, published journals and re-search contents. The survey revealed that in the field of traditional Chinese medicine (TCM), the number of sec-ondary research of patent literature was on the increase, and the degree of concern of TCM patent was far more than others. This survey not only assisted the understanding of current condition of TCM patent literatures, but also held its development trend in order to apply it in the studies.

Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.

We have designed the fuzzy associative memory pattern classifier (FAMPC) using multi-input and multi-output fuzzy set. It is adaptive to recognition of heart rate variability (HRV) signal, validity proved by many experiments.
Electrocardiography ; methods ; Fuzzy Logic ; Heart Rate ; Humans ; Pattern Recognition, Automated ; methods ; Signal Processing, Computer-Assisted

Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.
Animals ; Bronchi ; metabolism ; Calcium ; metabolism ; Calcium Channels ; Calcium Signaling ; physiology ; Cells, Cultured ; Male ; Membrane Glycoproteins ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; ORAI1 Protein ; Protein Kinase C-delta ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stromal Interaction Molecule 1

Objective To find humoral protein markers to develop new experimental diagnostic methods for tuberculous pleurisy. Methods Proteomes of 7 d and 14 d culture filtrate of Mycobacterium tuberculosis growing in Middlebrook 7H9, serum and pleural effusion from five patients with tuberculous pleurisy were detected by surface-enhanced laser desorptionionization time-of-flight massspectrometry (SELDI-TOF-MS). And the data were analyzed with descriptive statistics method to observed the protein components all owned by three kinds of proteome. Results From protein species, proteins of all culture filtrate were far more than that of pleural effusion and serum while proteins of pleural effusion in four cases were more than that of serum. The kinds of common proteins between culture filtrate and pleural effusion, between culture filtrate and serum, between serum and pleural effusion, among culture filtrate and pleural effusion and serum were different. But the protein of relative molecular mass of 2 660 depending on the ratio of mass to charge existed in all samples of culture filtrate, pleural effusion and serum. Conclusion The protein of relative molecular mass of 2 660 possess the latent quality as a specific humoral protein marker for tuberculous pleurisy. But it is essential that must be further confirmed among large samples.

Objective To evaluate the efficacy and safety of single flexible ureteroscope vs.flexible ureteroscope combined with intraoperative ultrasonography guided in the endogenous renal cyst incision and drainage.Methods Retrospective analysis of case data of 64 patients with endogenous renal cysts admitted from January 2015 to December 2017.All the patients undrwent contrast-enhanced CT,urinary tract imaging,ultrasound B to confirm bosniak classification Ⅰ and Ⅱ renal cyst.The patients (38 males,26 females) were divided into single flexible ureteroscope group [group A,32 cases of patients,average age (54.2 ±6.6) years,unilateral or bilateral cystic patients:19 cases/ 13cases,simple cyst of kidney or polycystic kidney:24 cases/8 cases,average maximum diameter of the renal cyst (5.4 ± 1.3) cm,combined with renal calculus:8 cases,combined with hydronephrosis:7 cases,1 case who had the history of laparoscopic renal cyst decapitation,1 case who had the history of percutaneous renal cyst puncture] and flexible ureteroscope group combined with intraoperative ultrasonography guided [group B,32 cases of patients,average age (52.3 ± 9.3) years,unilateral or bilateral cystic patients:21 cases/11 cases,simple cyst of kidney or polycystic kidney:25 cases/7 cases.Average maximum diameter of the renal cyst (5.3 ±1.2) cm,combined with renal calculus:9 cases,combined with hydronephrosis:8 cases,2 case who had the history of laparoscopic renal cyst decapitation,1 case who had the history of percutaneous renal cyst puncture].The double J stent was placed two weeks before the surgery into the body of patients.During the operation,the surgical procedure were used by the electronic flexible ureteroscope and holmium laser (0.8 J,20 Hz),combined with ultrasound B in the monitoring,to find the suitable incision position.It would be defined as the successful result if we could put the head of flexible ureteroscope into the inside of renal cyst and see the image of mist in the screen of ultrasound B.According to the standard,the result of operation wound be defined compared to before operation (success:shrink more than 80％ or maximum diameter was smaller than 1 cm;improvement:shrink about 50％-79％;ordinary result:shrink about 30％-49％;failure:shrink smaller than 30％).Effective rate =(success cases + improvement cases)/total cases.We compared the differences of two groups in the time,efficacy,safety and complication of surgery.Results Compared the results of single flexible ureteroscope groups vs flexible ureteroscope group combined with intraoperative ultrasonography guided in the success cases of surgery,after 8-30 months following-up,total operative complication rate was [8 cases(25％)vs.1 case(3.1％)],success rate of operation in 6 month later [(20 cases (62.5％) vs.28 cases (87.5％)].Flexible ureteroscope group combined with intraoperative ultrasonography guided had advantages statistically (P ＜ 0.05).However,there was no statistical difference for the two groups in the time of operation [(33.4 ± 6.7) min vs.(35.1 ± 8.6) min],single operative complication (intraoperative wounding bleeding:3 cases vs.1 case,postoperative urinary infection:3 cases vs.0 case,perinephric space effusion:2 cases vs.0 case,renal function impairment:1 case vs.0 case)(P ＞0.05).Conclusions Flexible ureteroscope combined with intraoperative ultrasonography guided can help the urologist can improve the success rate of surgery and reduce the chance of surgical injury intraoperatively.

Objective:To explore the application value and initial clinical experience of fluorescence-enhanced laparoscopy combined with indocyanine green in radical cystectomy with pelvic lymphadenectomy.Methods:A retrospective analysis was performed on the clinical data of a total of 8 patients admitted from May 2018 to August 2019 who underwent laparoscopic radical cystectomy with pelvic lymphadenectomy for muscle-invasive bladder cancer using the fluorescent laparoscopy system.There were 6 males and 2 females; aged 58 to 71 years, with an average of 65.8 years. The preoperative pathology was urothelial carcinoma, with 2 cases in T 2a stage, 4 cases in T 2b stage, and 2 cases in T 3a stage. The patient was injected a total of 2 ml of 2.5 mg/ml of indocyanine green through a rigid cystoscope using a 18 gauge cystoscope injection needle before surgery. The PINPOINT endoscopic fluorescence camera system performed surgery in real-time. A systematic examination of the surgical area under near-infrared fluorescence was performed every 5 minutes. Observation and recording of indocyanine green combined with fluorescence laparoscopy to mark the bladder tumor area and identify the sentinel drainage. Results:Eight cases were successfully completed under the fluorescence laparoscopic system.In terms of tumor markers, cystoscopy with indocyanine green allowed 7 of the 8 patients to successfully identify the tumor area under fluorescent laparoscopy. Six of the 8 patients with sentinel lymph nodes identified by indocyanine green lymphangiography were also successful, of which 3 were positive for lymph nodes. At the same time, intravenous indocyanine green was used to observe that the blood supply to the bladder and the end of the ureter was good. The operation time was 190-310 minutes, with an average of 235 minutes; the bleeding volume was 150-380 ml, with an average of 180 ml; the postoperative hospital stay was 10-22 days, with an average of 14.5 days. Postoperative pathological staging 2 cases in T 2a stage, 3 cases in T 2b stage, 3 cases in T 3a stage, with total number of lymph nodes 8-14, average 10. The 8 cases were followed-up for 4-12 months, average 5.5 months, without indocyanine green related complications. Conclusions:Indocyanine green fluorescent laparoscopy can accurately locate and identify the bladder tumor area and pelvic lymph nodes during radical cystectomy, which can reduce positive margins and unnecessary enlarged lymph node dissection.

Objective:To analyze the epidemiological and clinical characteristics of children with brucellosis, and to provide a practical basis and experience for clinical diagnosis and treatment of brucellosis.Methods:Retrospective analysis was used to collect clinical data of children with brucellosis diagnosed at the Children's Hospital Affiliated to Zhengzhou University from May 2015 to October 2017, and their epidemiological characteristics, clinical manifestations, laboratory tests, and clinical diagnosis and treatment were summarized.Results:A total of 24 children were included, including 14 males and 10 females, with an average age of 6 years (1 year 2 months to 12 years old). Except February, onset throughout the year, higher incidence was from May to July (14 cases, 58.33%). The exposure history of the children was mainly exposure to cattle and sheep and consumption of beef and mutton (18 cases, 75.00%). The main clinical manifestations were fever in 24 cases (100.00%), bone and joint pain in 14 cases (58.33%), hepatomegaly in 9 cases (37.50%), splenomegaly in 7 cases (29.17%). Tube agglutination test (SAT) was positive in 21 cases (87.50%), weakly positive in 1 case (4.17%) and negative in 2 cases (8.33%). Brucella was detected in all 24 cases by microbial culture, and in 18 cases (75.00%) by blood culture. Eighteen cases (75.00%) had liver dysfunction. Thirteen cases were misdiagnosed, and the misdiagnosis rate was as high as 54.17%. Twenty-two cases had been cured after treatment, 2 cases relapsed and recovered after continued treatment. Conclusions:Children with brucellosis have diverse epidemiology and clinical features, and are easily misdiagnosed. For children with fever, bone and joint pain and exposure history, pediatricians should be alert to the possibility of brucellosis, conduct microbiological and serological tests, in order to timely, accurate and standardized diagnosis and treatment of children with brucellosis.

Cerebrovascular disease is one of the main diseases that threaten human health and life,and the age of onset become younger and younger. Research shows that Toll-like receptors (TLRs) express in brain, have effects on neurons, play an important role in the occurrence and development of cerebral ischemia and secondary cerebral injury.It is clear that TLR2, TLR3, TLR4, TLR7 and TLR9 have influence on occurrence and development of cerebral ischemia. This is crucial for understanding the relationship between immune inflammation and nerve regeneration, and improving clinical treatment.