Objective To investigate the pathogenesis of high-altitude cerebral edema(HACE)and develop new therapeutic strategies.Methods Male Sprague-Dawley(SD)rats of 6 weeks old were selected and placed in a hypobaric chamber.The rats were exposed to the high-altitude environment of 7000 m above sea level for 3 days for HACE modeling.Whether the HACE model was successfully established in the rats was evaluated by measuring brain water content,the degree of disruption to the blood-brain barrier(BBB),and brain tissue Nissl staining.The experimental animals were divided into four groups,with 28 rats in each group.The blank control group was exposed to a normobaric and normoxic environment simulating the conditions at 500 m above sea level for 3 d.The other groups,including a model group(the HACE group),a bumetanide group(the positive control group),and a XH-6003 treatment group,were placed at an altitude of 7 000 m above sea level and were injected with normal saline,bumetanide,and XH-6003,a new type of Na-K-2C1 cotransporter 1(NKCC1)inhibitor,via the tail vein,respectively,twice daily for 3 d.The experimental animals were taken out of the hypobaric chamber for testing after 3 d.The primary outcome measures included brain water content,BBB permeability,changes in brain tissue morphology,and the expression levels of aquaporin-4(AQP4)and NKCC1.The secondary outcome measures included behavioral changes,apoptosis,and oxidative stress markers.Results The HACE rat model was successfully established.The model group exhibited increased brain water content(P＜0.0001),BBB disruption(P＜0.0001),impairment in learning skills and memory(P＜0.001),and anxiety/depression-like behaviors(P＜0.01).qPCR results showed significantly increased expression of NKCC1 and AQP4 in the brain tissue of the model group(P＜0.01).Pathology examination revealed neuronal and glial cell damage in the hippocampus of the model group(P＜0.01).Treatment with XH-6003,the NKCC1 inhibitor,reversed brain water content,BBB disruption,and neuronal and glial cell damage to a certain degree(P＜0.05),decreased the expression of NKCC1 and AQP4 in the brain tissue(P＜0.01),and inhibited apoptosis-related proteins.Among the oxidative stress indices,only glutathione(GSH)showed improvement(P＜0.001).Rats treated with XH-6003 showed functional improvement only in the time spent exploring novel objects,while other behavioral outcomes remained unchanged.Conclusion HACE is associated with the activation of the NKCC1/AQP4 pathway.Inhibition of this pathway alleviates brain edema,BBB disruption,and neuronal and glial cell damage.These findings suggest that XH-6003 holds potential as a therapeutic strategy for HACE at the cellular and molecular levels,but its effects in improving HACE-related behavioral disorders warrant further investigation.

The incidence of colonic diverticular disease is increasing significantly and is showing a trend of youthfulness.The treatment of colonic diverticulum disease mainly involves drugs,including antibiotics,rifaximin,and mesalazine.In addition,probiotics and dietary fiber are believed to have a positive impact on the treatment of diverticular diseases.When it progresses to acute complicated diverticulitis,surgical treatment becomes a necessary means.Up to now,our country has not formulated standards and guidelines of the diagnosis and treatment for colonic diverticulitis.

OBJECTIVE：To establish the metho d for the concentration det ermination of foretinib derivative LWK- 126 in liver microsomes，and to study its metabolism stability in liver microsomes of rats ，Beagle dogs and human. METHODS ：In the in vitro incubation system of liver microsomes ，LWK-126 was dissolved in liver microsomal incubation systems of rats ，Beagle dog and human initiated by reduced nicotinamide adenine dinucleotide phosphate solution. After incubation in water at 37 ℃ for 0，5，10，20， 30 and 60 min，the reaction was terminated with acetonitrile containing internal standard （1 μg/mL tolbutamide）. UPLC-MS/MS method was applied to determine the concentration of LWK- 126 in the incubation systems. The determination was performed on Waters BEH C 18 column with mobile phase consisted of water （containing 0.1％ formic acid ）-acetonitrile（containing 0.1％ formic acid）by gradient elution at the flow rate of 0.4 mL/min. The column temperature was 30 ℃，and the sample size was 2 μL. The mass spectral analysis was performed in a positive electrospray ionization mode ，and the full MS experiment was run with the selective reaction monitoring mode with a scanning range of m/z 50→1 200. Taking the concentration of LWK- 126 at 0 min as reference，the remaining percentage and the enzyme kinetic parameters were calculated. RESULTS ：The linear range of LWK- 126 was 0.05-15 μg/mL（R 2＝0.999 2）；the lower limit of quantification was 0.05 μg/mL，and the lowest detection limit was 0.01 μg/mL. The precision，accuracy，extraction recovery and matrix effect all met the analysis requirements of biological samples. The remaining percentage of LWK- 126 in liver microsomes of human ，rats and Beagle dogs for 60 min were （33.17±4.52）％，（3.14± 6.73）％，（1.38±5.85）％；t1/2 of them were 33.15，11.76，5.62 min；the clearance rates were 38.45，118.81，245.76 μL（/ min·mg）， respectively. CONCLUSIONS ：The method for the content ； determination of LWK- 126 in liver microsomes is established successfully. The order of metabolic stability of LWK- 126 in 〔2016〕4015） liver microsomes of different species is human ＞rats＞Beagle dogs.

OBJECTIVE：To establish the method for determining protein binding rate of dipeptidyl peptidase- 4 inhibitor LGT- 6 in different species of plasma ，and to compare their difference. METHODS ：By equilibrium dialysis ，LGT-6（3，30，300，3 000 nmol/L）was equilibrated in rat ，monkey and human plasma （i. e. internal dialysis solution ）for 48 h，using phosphate buffer as the external dialysis solution. The concentration of LGT- 6 in internal and external dialysis solution was determined by UPLC-MS/MS using tolbutamide as internal standard ，and the plasma protein binding rate was calculated. The determination was performed on ACQUITY UPLC HSS T 3 column with water （containing 0.01％ formic acid ）-acetonitrile（containing 0.01％ formic acid ）as mobile phase at the flow rate of 0.6 mL/min. The column temperature was 40 ℃，and the sample size was 2 μL. The ion source was electrospray ion source ，and the multiple ion monitoring mode was used to carry out positive ionization scanning. The ion pairs for quantitative analysis were m/z 487.0→434.3（LGT-6），m/z 271.1→172.0（internal standard ），respectively. RESULTS ：At the concentrations of 3，30，300，and 3 000 nmol/L，the protein binding rates of LGT- 6 in rat plasma were （96.25±0.97）％，（84.16± 1.24）％，（78.25±0.61）％，（66.63±0.95）％；the protein protein binding rates in monkey plasma were （98.54±0.58）％，（87.27± 1.01）％，（79.35±0.86）％，（66.69±0.54）％；the protein binding rates in human plasma were （99.40±1.03）％，（84.48± 1.15）％，（77.62±0.77）％，（66.93±0.48）％. At the same concentration ，the protein binding rates of LGT- 6 in rat ，monkey and human plasma had no significant difference （P＞0.05）. In the same species of plasma ，there were significant differences in the plasma protein binding rates of different concentration of LGT-6 among those groups （P＜0.05），and it decreased with 才〔2016〕4015） the increase of drug concentration. CONCLUSIONS ： The method for the determination of plasma protein binding rate of LGT-6 is successfully established. The data revealed that the protein binding rate of LGT- 6 is concentration-dependent ， there was no obvious spec ies difference on protein binding rates of LGT- 6 in rat ，monkey and human plasma under the same concentration.

Objective:To establish an analysis method for determining the contents of 7 components including ganoderic acid C2, ganoderic acid B,ganoderic acid G,ganoderic acid A,lucidenic acid A,ganoderma D and ganoderic acid F in ganoderma by QAMS. Methods:An HPLC method was used. The column was WatersX-bridge C18(250 mm × 4.6 mm, 5 μm), the mobile phase was acetonitrile(A)-1% acetic acid(B) (28:72) with gradient elution(0-30 min:A-28% to 38%;30-45 min:A-38% to 55%) at a flow rate of 1.0 ml·min-1,the column temperature was 30℃, and the detection wavelength was 254 nm. Ganoderic acid A was used as the internal reference, the correction factors of ganoderic acid C2, ganoderic acid B, ganoderic acid G, lucidenic acid A,ganoderma D and ganoderic acid F were calculated,and the contents of the 7 components were calculated by an external standard method to verify the feasibility and applicability of QAMS. Results:The relative correction factors of multiple assessments were reproducible,and the relative deviation of the contents of 6 components in 16 batches of Ganoderma lucidum was less than 2% when compared with that of the external standard method. Conclusion:QAMS is accurate and reliable in the quality evaluation of Ganoderma lucidum.

Objective To investigate the effect of ultrasound elastography combined with serum carbohydrate antigen 15-3(CA15-3 C), C keratin 19 fragment antigen 21-1(CYFRA21-1)and prostate specific antigen (PSA) levels detection on the sensitivity and accuracy of breast cancer diagnosis. Methods Sixty cases of breast cancer from December 2015 to January 2017 were selected as study group, and all cases were diagnosed as breast cancer by pathological examination.They were divided into groups according to the TNM classification criteria of breast cancer, stageⅠ22 cases, stageⅡ17 cases, stage Ⅲ14 cases, stageⅣ7 cases.Fifty-three cases of benign breast diseases were selected as control group. The levels of serum CA15-3, CYFRA21-1 and PSA were compared among two groups and the study group at different stages. The correlations between serums CA15-3, CYFRA21-1 and PSA expression level and breast cancer staging were analyzed. The results of pathological examination were regarded as "gold standard". Serum CA15-3, CYFRA21-1, PSA levels, single detection of ultrasound elastography and combined detection of breast cancer results were compared. Results The levels of serum CA15-3, CYFRA21-1 and PSA in study group were higher than those in control group:(37.98 ± 4.71)kU/L vs.(8.58 ± 3.04) kU/L,(5.41 ± 3.24)μg/L vs.(1.42 ± 0.47)μg/L,(0.39 ± 0.11)μg/L vs. (0.16 ± 0.04) μg/L, and there were significant differences (P<0.01). The levels of serum CA15-3, CYFRA21-1 and PSA in patients with stageⅢandⅣof breast cancer were higher than those of patients with stageⅠandⅡof breast cancer,and there were significant difference(P<0.01).Spearman correlation analysis showed that serum CA15-3, CYFRA21-1 expression levels and breast cancer staging was positively correlated (P < 0.05). There was a negative correlation between serum PSA expression and breast cancer staging(P<0.05).The sensitivity 98.33%(59/60), specificity 96.23%(51/53)and accuracy rate 97.35%(110/113) of breast cancer detected by serum CA15-3, CYFRA21-1, PSA levels and ultrasonic elastography was higher than that of single dectction, and the difference was statistically significant (P < 0.05). Conclusions The levels of serum CA15-3, CYFRA21-1 and PSA are closely related to the occurrence and progression of breast cancer.The combination of serum CA15-3, CYFRA21-1 and PSA levels with ultrasonic elastography can improve the specificity, sensitivity and accuracy of diagnosis of breast cancer.

Objective To explore the diagnostic value of Talaromyces marneffei (T.marneffei)-specific mannose glycoprotein Mp1p antigen for T.marneffei infection in acquired immune deficiency syndrome (AIDS) patients.Methods All cases were recruited in this study from January 2012 to June 2015 in Guangzhou No.8 People′s Hospital, including 184 AIDS patients with T.marneffei infection confirmatively diagnosed by culture, and 205 controls including 176 AIDS patients without T.marneffei infection and 29 health controls.Double antibody sandwich enzyme linked immunosorbent assay and fluoroimmunoassay combined with double-antibody sandwich were both utilized to detect serum Mp1p antigen levels, and their sensitivity and specificity for diagnosing T.marneffei infection in patients with AIDS were analyzed.x2 test and t test were used for statistical analysis.Results The ratio of males to females and age of the study group were both comparable to those of the control group (x2=0.019, P=0.889;t=1.810,P=0.07, respecitvley).The sensitivities of double antibody sandwich enzyme linked immunosorbent assay and fluoroimmunoassay combined with double-antibody sandwich were 82.07%(151/184) and 83.15%(153/184), respectively (x2=0.076, P=0.783).The specificities were 93.17%(191/205) and 92.68%(190/205), respectively (x2=0.037, P=0.847).The accuracy values were 87.92%(342/389) and 88.17%(343/389), respectively (x2=0.012, P=0.912).The false positive rates were 6.83%(14/205) and 7.32%(15/205), respectively.The false negative rates were 17.93%(33/184) and 16.85%(31/184), respectively (x2=0.049, P=0.829).The positive predictive values were 91.52%(151/165) and 91.07%(153/168), respectively (x2=0.021, P=0.886).The negative predictive values were 85.27%(191/224) and 85.97%(190/221), respectively (x2=0.045, P=0.832).The Kappa values were 0.83 and 0.80, respectively.Conclusion Detection of serum Mp1p antigen of T.marneffei possesses high specificity and sensitivity, which may be utilized for rapid and early diagnosis of T.marneffei infection in patients with AIDS.

Objective: To establish a quantitative determination method for β-eudesmol in Cortex Magnoliae Officinalis by GC. Methods:β-Phenethanol was used as the internal standard substance;the column was a Zebron ZB-WAX capillary column ( 60 m × 320 μm,0. 5μm) with the column temperature of 200℃;the detector was FID and the vaporizer temperature was 250℃; the carrier gas was nitrogen with the flow rate of 1. 3 ml · min-1 and the split ratio was 4 ∶1. Results: The linear range of β-eudesmol was 0. 015 1-0. 271 2 mg·ml-1(r=0. 999 8);the average recovery was 99. 28%(RSD =1. 17%, n=6). Conclusion:The method is simple and accurate with good reproducibility, which can be used for the quality control of medicinal materials and decoction pieces of Cortex Magnoliae Officinalis.

Objective:To study the changes and significance of Th17 and Treg cells as well as their associated cytokines in pe-ripheral blood of patients with chronic hepatitis C and cirrhosis in order to investigate their role in the pathogenesis of chronic hepatitis C and hepatitis C cirrhosis. Methods:Flow cytometry and ELSIA assay were used to detect the expression ratio of Th17 and Treg cells and the serum levels of IL-10,TGF-β,IL-6 and IL-17 in peripheral blood of healthy human and patients with chronic hepatitis C and hepatitis C cirrhosis. Than we analyzed the differences of the above detection index between the healthy and patients with chronic hepatitis C and hepatitis C cirrhosis. Results:The expression ratio of Th17 cells (1. 33%±0. 30%) in peripheral blood of patients with chronic hepatitis C and IL-6[(8. 10±2. 42)ng/L] and IL-17[(16. 70±4. 73)ng/L] serum levels were significantly higher than those in the healthy (Th17:1. 14%±0. 19%) and IL-6[(1. 72±6. 70)ng/L],IL-17[(12. 29±1. 88)ng/L],P<0. 05;The expression ratio of Treg cells in peripheral blood of patients with hepatitis C cirrhosis and chronic hepatitis C patients was significantly higher than that in the healthy (6. 21%±0. 76%,5. 89%±0. 85% vs 5. 51%±0. 59% ),P<0. 05,The ratio of Th17/Treg in patients with cirrhosis was significantly lower than that in patients with chronic hepatitis C and the healthy(0. 19±0. 02 vs 0. 22±0. 03,0. 21±0. 03),P<0. 05;the levels of IL-1[(16. 21±3. 76)ng/L] and TGF-β[(5. 15±0. 83)ng/L] in peripheral blood of patients with hepatitis C cirrhosis were significantly higher than those in chronic hepatitis C[(14. 36±2. 78)ng/L;(4. 47±0. 87)ng/L] and the healthy[(14. 01±3. 01)ng/L;(4. 43±0. 98)ng/L)],P<0. 05. Conclusion:Th17 and Treg cells and their related cytokines involved in the process of chronicity of hepatitis C and cirrhosis,Th17 and Treg cells and their related cytokines in the treatment and prevention of chronicity of hepatitis C and cirrhosis may have important significance.

Objective:To evaluate the quality of Dendrobii officinalis Caulis cultivated in Lishui by the content determination of ex-tract, polysaccharide and mannose. Methods:Totally 26 batches of Dendrobii officinalis Caulis were dried at 55℃, and the ethanol ex-tract, polysaccharide and mannose was determined respectively by hot dipping, phenol-sulphuric acid colorimetry and pre-column deri-vatization HPLC method according to the determination methods for Dendrobii officinalis Caulis recorded in Chinese Pharmacopoeia (2010 edition). Results:The contents of extract, polysaccharide and mannose in Dendrobii officinalis Caulis during the collection pe-riod were higher than those of the pharmacopoeia standard, and there were significant differences among the batches. Conclusion:The test can provide theory basis for the quality evaluation of Dendrobii officinalis Caulis cultivated in Lishui, and provide guidance for the planting of the herb.