To establish a method for detecting microdialysis recovery of tetramethylpyrazine (TMP) and ferulic acid (FA) and investigating the influencing factors, providing the basis for further in vivo microdialysis experiments. The concentration of FA and TMP in dialysates were determined by high pressure liquid chromatography ( HPLC) and probe recovery were calculated respectively. The influence of the flow rates, medium concentration, temperature and in vivo probe stability on the recovery of FA and TMP were investigated by using concentration difference method (incremental method and decrement method). The recovery obtained by incremental method were similar to by decrement method. The in vitro recovery rate of FA and TMP decreased with the increase of 1-2.5 μL min(-1), and increased obviously with the temperature of 25-42 degrees C under the same conditions. The concentration of FA and TMP had no obvious effect on the probe recovery under the same flow rate. In addition, the recovery of TMP and FA remained stable and showed similar trends under the condition of four concentration cycles, indicating that the intra day reproducibility of the concentration difference method was good. The recovery of brain microdialysis probes in vivo 8 h maintained a relatively stable, but certain differences existed between different brain microdialysis probes, demonstrating that each probe was required for recovery correction in vivo experiment. Microdialysis sampling can be used for the local brain pharmacokinetic study of FA and TMP, and retrodialysis method can be used in probe recovery of FA and TMP in vivo.
Animals ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; Coumaric Acids ; analysis ; isolation & purification ; pharmacokinetics ; Drugs, Chinese Herbal ; Humans ; Microdialysis ; methods ; Pyrazines ; analysis ; isolation & purification ; pharmacokinetics ; Rats

OBJECTIVETo study the expressions of gastric mucosal proteins in chronic gastritis (CG) rats of Pi-Wei damp-heat syndrome (PWDHS), to investigate the pathogenesis correlated to CG rats of PWDHS, to observe the differential expressions of gastric mucosal proteins in CG rats of PWDHS, and to investigate the mechanisms of Sanren Decoction (SD) for treating CG rats of PWDHS.

METHODSTotally 36 male SD rats were adaptable fed for 3 days and randomly divided into 3 groups, i.e., the normal control group, the CG of PWDHS rat model group (abbreviated as the model group), and the SD treatment group, 12 in each group. The CG of PWDHS rat model was prepared by composite factors. The gastric mucosal protein was separated using two-dimensional gel electrophoresis technique, and stained by Coomassie brilliant blue. The protein spots expressed differently were analyzed by PDquest 8.0 software. The protein spots expressed differently was identified by MALDI-TOF/TOF-MS.

RESULTSThe protein spots were 1 025 +/- 3 9, 994 +/- 51, 1 087 +/- 33 in the normal control group, the model group, and the SD treatment group respectively detected from two-dimensional gel electrophoresis profiles. Compared with the normal control group, there were 74 protein spots differentially expressed in the model group, 30 spots up-regulated and 44 spots down-regulated. Compared with the model group, there were 75 protein spots differentially expressed in the SD treatment group, 49 spots up-regulated and 26 spots down-regulated. Five protein spots differentially expressed were successfully identified, i.e., heat shock protein 72 (HSP72), heat shock protein 60 (HSP60), protein disulfide-isomerase (PDI), malate dehydrogenase (MDH), and unnamed protein.

CONCLUSIONSThe pathogenesis of CG of PWDHS may be correlated to energy metabolism disturbance and stress. The mechanisms of SD for treating it may possibly adjust differential expressions of gastric mucosal proteins.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; metabolism ; Gastritis ; diagnosis ; drug therapy ; metabolism ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Proteome ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective:To study the impact on dose accuracy for the treatment planning by manually assigning accurate electron density for CT image-based tumor tissues and organs at risk.Methods:Twenty cases of retrospective postoperative cervical cancer radiotherapy plans were selected. The body electron density of the corresponding organs was derived from the ICRU 46 report and assigned in the treatment planning system (Monaco5.11, Sweden), including the bladder, rectum, intestine, kidney, spinal cord, femoral head, and ilium. The original plans were double-arc volumetric modulated arc therapy plan (360° VMAT), using Monte Carlo algorithm, the calculation grid was 0.3 cm × 0.3 cm × 0.3 cm, and the minimum subfield width was 0.6 cm. Keep the original plan fluence unchanged and recalculate the dose to generate a new plan. The two-dimensional dose distribution and dose-volume histogram (DVH) were used to compare the differences between the two plans. The difference was compared between the two group plans by using the dosimetry parameters and DVH two dimension curve.Results:For the planning of assigning bulk electron density (Plan RED), the deviation of the patient′s target dose parameters and the original plan (Plan ref) was <2%, and the average deviation of all target regions D2, D98, Dmean was < 0.7%, only 2 of the 180 data were between 2% and 3%. The average deviation of V20, V30, D1 cm 3, Dmean of the bladder, rectum, and small intestine, the original Plan ref was less than 0.6%, and 4 out of 240 data had values > 2%. Plan RED′s average hop count was 0.9% higher than Plan ref, and the total number of subfields remains unchanged. The planned dose generated by manually assigning the electron density in Plan RED was higher than that in Plan ref, but met the clinical requirements. The two-dimensional curves of the DVH diagram for targets and OARs almost completely overlapped, and there was no obvious difference in the dose distribution diagram of the same cross section. The statistical result of all parameters showed that the difference in planned dose parameters between the two groups was not statistically significant( P>0.05). Conclusions:The overall deviation of dose accuracy between Plan RED and Plan ref is <2%, which meets the clinical requirements and provides a reference for realizing MRI-only treatment planning.

Nowadays,surgery is a comprehensive approach for the treatment of hepatic carcinoma as the first choice,but there are still many limitations in surgical therapy,such as the location,boundary and metastasis of hepatic carcinoma.Currently,the indocyanine green fluorescence imaging-guided hepatectomy is widely used at home and abroad as a new progress and hotspot in hepatobiliary surgery,which makes hepatobiliary surgery more convenient and makes up for some deficiencies in hepatectomy.Thus,we summarized the experience of indocyanine green fluorescence imaging-guided hepatectomy in the People's Hospital of Hunan Province and discussed its value of application.

OBJECTIVETo study the role of O(6)-methylguanine-DNA methyltransferase (hMGMT) in the development of human lung cancer.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) method was applied to measure hMGMT mRNA expression in 150 lung cancer specimens, 40 normal lung tissues, and in the peripheral mononuclear blood cells from 50 lung cancer cases and 50 normal controls. The protein expressions of p53, C-MYC and K-RAS were assessed by immuno-histochemistry. The effects of some exposure factors on the expression of hMGMT gene were analyzed. The relationships between hMGMT gene and cancer related genes p53, C-MYC and K-RAS were investigated.

RESULTSThe mRNA of hMGMT was low or absent in 49 of 150 (32.7%) lung cancer specimens, whereas 2 of 40 (5%) normal lung tissues had reduced the levels of hMGMT mRNA. The low expression of hMGMT seemed to be a risk factor of lung cancer, with a OR of 9.22 (2.05-57.65). Reduced expression levels of hMGMT mRNA were observed in 10 of 50 (20%) lung cancer patients' peripheral mononuclear blood cells, and 2 of 50 (4%) blood cells among normal controls. When investigating the exposure factors which affecting the expression of hMGMT gene, we noticed that smoking was suppressing the expression of hMGMT gene. Interestingly, over-expression of K-RAS oncogene was significantly correlated with low expression of hMGMT (P < 0.05). However, the expressions of p53 and C-myc were not correlated with the status of hMGMT gene.

CONCLUSIONhMGMT might play an important role in the development of human lung cancer. Low expression of hMGMT gene seemed to be a risk factor for lung cancer which could be used as a valuable biomarker on susceptibility of human lung cancers.

Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Squamous Cell ; enzymology ; genetics ; China ; epidemiology ; DNA Repair ; genetics ; Female ; Genes, ras ; genetics ; Humans ; Lung Neoplasms ; enzymology ; genetics ; Male ; Middle Aged ; O(6)-Methylguanine-DNA Methyltransferase ; biosynthesis ; genetics ; metabolism ; Point Mutation ; RNA, Messenger ; biosynthesis ; genetics ; Smoking ; adverse effects ; ras Proteins ; biosynthesis ; genetics

OBJECTIVETo analyze the distribution of trimethylamine N-oxide (TMAO) in healthy adults with different risk factors and explore its association with gut microbiota.

METHODSWe collected fasting blood samples and fresh fecal samples from 181 subjects without atherogenesis in the carotid arteries. Plasma TMAO levels of the subjects were determined using stable isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS). The fecal DNA was extracted, and the 16S rRNA V4 tags were amplified and sequenced by Illumina HiSeq 2000. The association between TMAO and classical cardiovascular risk factors were analyzed. Gut microbial community structure was analyzed with QIIME, and LEfSe was used to identify the biomarkers.

RESULTSThe median (IQR) TMAO level was 2.66 (1.96-4.91) µmol/L in the subjects. TMAO level was significantly correlated with body mass index and operational taxonomic units (OTU). Individuals with high TMAO levels were found to have abundant Clostridiales, Phascolarctobacterium, Oscillibacter, and Alistipes but less abundant Anaerosprobacter.

CONCLUSIONChinese subjects have in general low levels of TMAO. TMAO levels are not significantly correlated with the classical cardiovascular risk factors or the gut microbial structures.

Adult ; Atherosclerosis ; Bacteria ; isolation & purification ; Biomarkers ; blood ; Cardiovascular Diseases ; blood ; Chromatography, Liquid ; Gastrointestinal Microbiome ; Humans ; Methylamines ; blood ; RNA, Ribosomal, 16S ; isolation & purification ; Risk Factors ; Tandem Mass Spectrometry

OBJECTIVETo evaluate the effect of short-course kidney-invigorating therapy on near-term semen quality in asthenozoospermic men with kidney deficiency.

METHODSBased on the differential types in traditional Chinese medicine, 121 asthenozoospermia patients received at our clinic of andrology were divided into groups A (kidney-yin deficiency), B (kidney-yang deficiency) and C (spleen and kidney deficiency), and treated with Yougui Decoction plus Wuziyanzong Pills, Jinkuishenqi Pills plus Wuziyanzong Pills, and Shizi Decoction plus Liujunzi Decoction, respectively, all given once daily for 4 weeks. Sperm parameters of the patients were analyzed with the computer-assisted sperm analysis system before and after treatment and compared among the three groups.

RESULTSThe baseline sperm concentrations in groups A, B and C ([70.4 +/- 38.6], [73.5 +/- 40.2] and [56.0 +/-34.4] x 10(6)/ml) showed no significant differences from those after medication ([74.4 +/- 32.6], [67.0 +/- 30.8] and [58.6 +/- 24.6] x 10(6)/ml) (P > 0.05). The percentages of grade a sperm in the three groups were (12.9 +/- 5.3)%, (13.7 +/- 7.7)% and (12.9 +/- 6.4)% respectively after treatment, significantly higher than (9.9 +/- 6.7)%, (9.3 +/- 5.4)% and (9.0 +/- 6.8)% before treatment (P < 0.05), and so were the percentages of grade a + b sperm ([37.4 +/- 10.2 ]%, [35.7 +/- 13.7]% and [35.9 +/- 12.3]% after treatment versus [29.6 +/- 13.2]%, [27.5 +/- 10.4]% and [28.3 +/- 12.1]% before treatment, P < 0.05). All the three groups showed significantly increased sperm motility after treatment ([53.8 +/- 10.5]%, [52.6 +/- 15.2]% and [51.1 +/- 13.1]%) as compared with the baseline levels ([44.3 +/- 14.0]%, [43.5 +/- 15.0]% and [42.4 +/- 14.9]%) (P < 0.05). The cure rate and total effectiveness rate were significantly higher in group B than in A (P < 0.05), but had no significant differences between either A and C or B and C (P > 0.05).

CONCLUSIONShort-course kidney-invigorating therapy can significantly improve near-term semen quality in asthenozoospermic men with kidney asthenia, especially in those with kidney-yang deficiency, and it has no obvious adverse effects.

Adult ; Asthenozoospermia ; diagnosis ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Oligospermia ; diagnosis ; drug therapy ; Phytotherapy ; Semen Analysis ; Yang Deficiency ; Young Adult

Objective To summarize risk factors for clinical outcomes in heart transplantation patients, evaluate the characters of Chinese patients by comparing with international data, and introduce new clinical strategies. Methods We performed 200 heart transplantations from Jun. 2004 to May 2010. The clinical information was recorded and all patients were followed up. By analyzing 160 patients with a follow-up period of more than one year, we summarized clinical outcomes and risk factors of early and late results of heart transplant patients. Results Of 160 patients, 8. 1 % received postoperative extracorporeal membrane oxygenation (ECMO) support and 10% continuous renal replacement therapy. In 550 cases/times of endomyocardial biopsies, the incidence of rejection with grades more than Ⅱ (concluding grade Ⅱ ) was 14. 9%. In-hospital mortality was 3. 8%. Smoking,preoperative diastolic pulmonary arterial pressure, PAWP, total serum protein level and albumin level were risk factors of peri-operative mortality, and preoperative diastolic pulmonary arterial pressure,primary heart diseases, pulmonary hypertension and implantations of ICD, MCS and ECMO were risk factors of late mortality. Postoperatively, 1-, 3- and 5-year survival rate was 94. 4%, 91.9% and 88. 8%, respectively. Compared with UNOS data, the rate of primary heart diseases, pulmonary hypertension, and implantation of ICD, MCS and ECMO were different, and the long-term survival rate of 160 patients was higher than that reported by ISHLT. Conclusion The risk factors of mortality of Chinese heart transplant patients are different with their counterparts from western countries. Our corresponding peri-operative treatments and clinical strategies have produced satisfactory clinical outcomes.

Objective To investigate the clinical features of patients with bronchiectasis of different types.Methods One hundred and twenty two patients with bronchiectasis at stable stage were recruited from January 2014 to July 2015.The patients were typed as cystic bronchiectasis (n =45) or non-cystic bronchiectasis (n =77) by high resolution CT (HRCT),expectoration bronchiectasis (n =80) or dry brochiectasis (n =42) by clinical symptoms,bacterial colonization (n =42) or non-bacterial colonization (n =80) by sputum culture.The modified British Medical Research Council (mMRC) dyspnea scale,Leicester Cough Questionnaire (LCQ),St George's Respiratory Questionnaire (SGRQ) and pulmonary function test were used to assess the clinical features,and the episodes of exacerbations and hospitalization,and mortality during 1-year follow-up were documented.Results mMRC dyspnea scale (1.90 ± 0.94 vs.2.90±1.09,t=-5.040),LCQ (16.20±4.60 vs.11.20±2.20,t=8.114),SGRQ (36.80±13.10 vs.52.06±22.10,t=-4.780),FEV1％ pred (68.45 ±26.50 vs.52.22 ±20.60,t=3.458),FVC％ pred (72.20 ±26.32 vs.63.10 ±21.42,t =2.058),FEV1/FVC (75.14 ±20.52 vs.58.12 ± 19.82,t =4.546),diffusing capacity of the lung for carbon monoxide (DLCO) (76.24 ± 28.40 vs.54.32 ± 21.20,t =4.400),episodes of exacerbations (Z =-8.272) and hospitalization during 1-year follow-up [6(14.29％) vs.29(36.25％),x2 =6.495] in patients with dry bronchiectasis were significantly better than those in patients with expectoration bronchiectasis (all P ＜ 0.05).mMRC dyspnea scale (3.20 ± 2.10vs.2.10±1.40,t=3.131),LCQ (10.12±2.63vs.16.22 ±3.22,t=11.365),SGRQ (54.80± 18.12 vs.34.06 ± 12.10,t =6.839) and FEV1％ pred (46.52 ± 22.55 vs.58.22 ± 24.62,t=-2.611),FVC％ pred (60.24± 18.22 vs.70.10±24.20,t =-2.547),FEV1/FVC (62.54± 19.02vs.73.12 ±18.42,t=-3.025),DLCO (62.24 ±22.40 vs.74.52 ±26.26,t=-2.627),episodes of exacerbations (Z =10.213) and hospitalizations during 1-year follow-up [21 (46.67 ％) vs.14 (18.18％),x2 =1 1.260] in patients with cystic bronchiectasis were significantly more severe than those in patients with non-cystic bronchiectasis (all P ＜ 0.05).mMRC dyspnea scale (2.38 ± 1.45 vs.1.92 ± 1.14,t =2.175),LCQ (12.82 ±2.12 vs.16.20 ±3.96,t =-6.140),SGRQ (54.22±21.50 vs.41.20 ± 14.60,t =3.521) and FEV1 ％ pred (54.20 ± 21.60 vs.66.45 ± 28.24,t =-2.668),FVC％ pred (63.10 ±24.32 vs.73.46 ±25.30,t =-2.177),FEV1/FVC (62.22 ±20.80 vs.72.14 ±24.36,t =-2.243),DLCO (58.52 ± 20.42 vs.69.22 ± 25.60,t =-2.344),episodes of exacerbation (Z =19.352) and hospitalization during 1-year follow-up [19 (45.24％) vs.16 (20.00％),x2 =8.575] in patients with bacterial colonization bronchiectasis were significantly more severe than those in patients with non-bacterial colonization bronchiectasis (all P ＜ 0.05).However,there was no significant difference in mortality during 1-year follow-up (all P ＞ 0.05) among patients with different types of bronchiectasis.Conclusion Patients with cystic,bacterial colonization and expectoration types of bronchiectasis seem to have more severe symptoms,more episodes of exacerbations and hospitalizations than those of non-cystic,non-bacteria colonization and dry types of bronchiectasis.

Objective To investigate the application value of two-step separation approach in laparoscopic hemihepatectomy.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 81 patients who underwent laparoscopic hemihepatectomy in the People's Hospital of Hunan Provincial between January 2015 and December 2017 were collected.Patients underwent laparoscopic hemihepatectomy using two-step separation approach after preoperative assessment.Hepatic pedicle,hepatic vein and branches were processed in the liver parenchyma,without intrathecal anatomy.Observation indicators:(1) preoperative assessment,intraand post-operative recovery;(2) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed up to February 2018.Patients with hepatolithiasis received abdominal CT reexaminations at 5-7 days postoperatively for observing postoperative residual stones,and patients with malignant tumor were regularly followed up for 1-3 years.Measurement data with normal distribution were represented as (x)±s.Count data were described as frequency and percentage.Results (1) Preoperative assessment,intra-and post-operative recovery:81 patients underwent preoperative blood biochemistry,tumor biomarker and imaging examinations,and remaining functional liver volume and a liver model in 9 patients were respectively calculated and built using the 3D reconstruction software and 3D printing technology.Of 81 patients,68 underwent left hemihepatectomy and 13 underwent fight hemihepatectomy;77 underwent successful laparoscopic hemihepatectomy using two-step separation approach,4 were converted to open operation,with a rate of 4.9％ (4/81).Of 4 patients with conversion to open operation,1 had difficult tumor separation due to tumor invading port vein induced to close adhesion,1 had stone removal difficulties under laparoscopy due to hepatolithiasis,and 2 were converted to open operation due to severe liver cirrhosis induced to massive intraoperative bleeding.Of 81 patients,70 gained dividing line of hemiliver by intraoperatively hemihepatic blood flow occlusion,and then got effectively control of bleeding combined with Pringle blood flow occlusion,and 11 received Pringle blood flow occlusion in whole liver.Laparoscopic fluorescence imaging technology was intraoperatively used for 2 patients.Operation time,volume of intraoperative blood loss,rate of intraoperative blood transfusion and duration of hospital stay in 81 patients were respectively (206±42)minutes,(195±134)mL,11.1％(9/81) and (11.5+2.7)days.Eighty-one patients were complicated with bile leakage and were cured by conservative treatment,with a bile leakage incidence of 2.5％ (2/81),and without severe complications,such as postoperative bleeding,hepatic dysfunction and subphrenic abscess.There was no perioperative death and reoperation within 30 days postoperatively.(2) Follow-up and survival situations:55 patients with hepatolithiasis were followed up and underwent CT examinations of upper abdomen at 5-7 days postoperatively,including 52 with depletion of stones;3 with residual stones received removal of stones by choledochoscope at 3 months postoperatively,without residual stones.Seventeen patients with malignant tumor were followed up for 12-36 months,with a median time of 15 months,16 had tumor-free survival,and 1 was complicated with intrahepatic metastasis at 1 year after resection of hepatocellular carcinoma,and then underwent transcatheter arterial chemoembolization (TACE) and survived with tumor.Nine patients with benign liver diseases had good recovery during follow-up.Conclusion Two-step separation approach that is rationally used in laparoscopic hemihepatectomy is safe,effective and convenient.