Objective:To establish a high-performance liquid chromatography(HPLC)method for the simultaneous determination of 10 bacteriostatic agents in Kangfuxin solution.Methods:The HPLC method parameters were as follows,a Thermo AcclaimTM 120 column(4.6 mm × 250 mm,5 μm),a gradient mobile phase of 0.1％glacial acetic acid-0.02 mol·L-1 ammonium acetate solution and methanol at a flow rate of 1 mL·min-1,and the scan-ning wavelength of 200-700 nm.Results:In the corresponding concentration range,the linear relationship be-tween the mass concentration of the 10 bacteriostatic agents and the peak area was good(r≥0.999 8);the preci-sion was good with RSD of 0.33％-0.38％;The RSD values of the repeatability test and the stability test were＜0.5％,the recovery rate was 97.4％-101.1％,and the RSD was between 0.4％-1.8％.According to the pro-posed method,the content results of benzoic acid,sorbic acid,hydroxyphenyl ethyl ester and hydroxyphenyl propyl ester in the test sample were 0.15-0.34％,0-0.48％,0-0.02％,and 0-0.02％,respectively.Conclu-sion:This method is sensitive,rapid,specific and accurate,and can be used for the determination of the content of bacteriostatic agents in Kangfuxin solution.

Objective:The expression of serum free light chain(FLC)is abnormal in various diseases,but its role in lung cancer remains unclear.This study aims to investigate the expression and diagnostic value of serum FLC in lung cancer. Methods:A total of 80 lung cancer patients treated at Xiangdong Hospital,Hunan Normal University from January to December 2021 were selected as the lung cancer group.Another 80 healthy individuals undergoing routine physical examinations during the same period were chosen as the control group.General information and serum κFLC and λFLC levels were collected for all subjects.Clinical indicators such as serum carcinoembryonic antigen(CEA),cytokeratin fragment antigen 21-1(CYFRA21-1)levels,tumor diameter,histological type,TNM stage,and lymph node metastasis status were recorded for lung cancer patients.The expression levels of serum FLC[κFLC,λFLC,and FLC(κ+λ)]were compared between the lung cancer group and the control group.Lung cancer patients were grouped based on gender,age,smoking history,tumor diameter,TNM stage,histological type,and lymph node metastasis to compare differences in serum κFLC and λFLC levels.Receiver operating characteristic(ROC)curves were used to evaluate the diagnostic value of serum FLC alone and in combination with other indicators in lung cancer. Results:The expression levels of serum FLC(κ+λ)and κFLC were significantly higher in the lung cancer group than those in the control group(both P＜0.001),while there was no significant difference in serum λFLC levels between the 2 groups(P＞0.05).There were no significant differences in serum κFLC levels among lung cancer patients with different tumor diameters,histological types,or TNM stages(all P＞0.05);however,serum κFLC levels were higher in lung cancer patients with lymph node metastasis than in those without,with statistical significance(P=0.033).There were no significant differences in serum λFLC levels based on tumor diameter or histological type(both P＞0.05),but serumλFLC levels were higher in stage Ⅲ+Ⅳ and lymph node metastatic lung cancer patients compared to stage Ⅰ+Ⅱ and non-metastatic patients,with statistical significance(P=0.033 and P=0.019,respectively).The area under the curve(AUC)for κFLC and CEA in diagnosing lung cancer showed no significant difference(P=0.333).The combination ofκFLC+CYFRA21-1 had the highest diagnostic efficacy(AUC=0.875)and sensitivity(71.3％).The AUC for the combined diagnosis of κFLC+λFLC+CEA+CYFRA21-1 was 0.915(95％CI 0.860 to 0.953,P＜0.001). Conclusion:Serum FLC is highly expressed in lung cancer and is associated with its invasion and metastasis.Serum FLC,particularly κFLC,has diagnostic value for lung cancer,and the combined detection of FLC,CEA,and CYFRA21-1 offers the best diagnostic efficacy.

Objective : To investigate the effect of Rotundic acid (RA) on proliferation，migration and invasion a- bility of human lung adenocarcinoma cells as well as its possible mechanisms． Methods : Human lung adenocarci- noma A549 and PC9 cells were divided into control group，blank control group，solvent group and 20，40，60，80 μmol / L RA groups．CCK-8 assay and scratch assay were used to detect the proliferation and horizontal migration of human lung adenocarcinoma cells．Transwell migration assay and Transwell invasion assay were used to detect the longitudinal migration and invasion ability of A549 and PC9 cells in each group．The protein expression levels of ja- nus kinase 2 (JAK2) and signal transducer and activator of transcription 3 ( STAT3) in the supernatants of A549 and PC9 cells were detected by ELISA．The mRNA expression levels of JAK2 and STAT3 were detected by RT- PCR. Statistical analysis was made on the differences among groups in each index. Results : After RA treatment on human lung adenocarcinoma cells ，compared with the control group ，the proliferation activity of A549 and PC9 cells in the experimental groups decreased (P＜0. 05) ，the number of cells crossing polycarbonate membrane and matrix glue decreased (P＜0. 05) ，the expression of JAK2 and STAT3 proteins in cell supernatant decreased (P < 0. 05) ，and the mRNA expression of JAK2 and STAT3 decreased (P＜0. 05) ．The decrease of the above indices was concentration-dependent and had statistical significance (P＜0. 05) ．Compared with the control group，the pro- liferation activity of A549 and PC9 cells in the solvent group showed no significant difference． Conclusion RA may inhibit the proliferation，migration and invasion of human lung adenocarcinoma A549 and PC9 cells in vitro， possibly through the inhibition of JAK2 / STAT3 pathway.

Objective To investigate the effects of skin/muscle incision and retraction(SMIR)on mechanical paw withdrawal threshold and the ability of spatial learning and memory in immature rats after adulthood. Methods 27 male SD rats aged 3 weeks and weighing 60 ~ 80 g were randomly divided into 3 groups(n = 9):control group(group C),sham operation group(group Sham)and skin/muscle incision and retraction group (group SMIR). Group SMIR received operation for skin/muscle incision and retraction. Sham group received skin/muscle incision but no retraction.No surgery was operated on C group. Pain behavior was assessed by mechanical paw withdrawal threshold(MWT)to von Frey filament stimulation before and 1,3,7,12,22 and 32 days after operation.The effects of spatial learning and memory function were assessed by Morris water-maze test at 33 days after operation. Results Mechanical paw withdrawal threshold of group SMIR decreased 1 day after operation (P<0.05)and showed no significant difference before and 3,7,12,22,32 days after operation in 3 groups(P >0.05). In Morris water-maze test,compared with Sham and C group,the average escape latency in SMIR was sig-nificantly longer in the water maze navigation experiment(P < 0.01);the ratios of time and path in the quadrant of the platform were obviously lower in SMIR(P < 0.01). There was no statistical difference between sham and C group(P>0.05).Conclusion SMIR did not cause chronic pain but may cause a decrease in the ability of spatial learning and memory in immature rats.

OBJECTIVEPsoriasis is an autoimmune-related chronic inflammatory skin disease strongly associated with the dysfunction of Th17 cells. Retinoic acid-related orphan nuclear receptor γt (RORγt) plays a critical role in the differentiation and maturation of Th17 cells and in cell-derived immunologic derangement. We conducted this study to investigate potential mechanism by which the derivative of digoxin selectively antagonizes RORγt transcriptional activity.

METHODUsing molecular docking in combination with molecular electrostatic potential (MEP), we detected the interaction between the derivative of digoxin (Dhd) and ROR transcription factor (RORα,RORβ and RORγt), and the results were further confirmed by bioluminescent assay.

RESULTMolecular docking demonstrated that Dhd could exclusively inhibit the conformation of RORγt; bioluminescent assay further indicated that RORγt was selectively antagonized by Dhd in a dose- and time-dependent manner.

CONCLUSIONDhd can selectively suppress RORγt transcriptional activity.

Digoxin ; analogs & derivatives ; pharmacology ; Humans ; Models, Chemical ; Molecular Docking Simulation ; Nuclear Receptor Subfamily 1, Group F, Member 1 ; antagonists & inhibitors ; genetics ; Transcription, Genetic

Objetive Psoriasis is an autoimmune-related chronic inflammatory skin disease strongly associated with the dysfunction of Th17 cells. Retinoic acid-related orphan nuclear receptorγt (RORγt) plays a critical role in the differentiation and maturation of Th17 cells and in cell-derived immunologic derangement. We conducted this study to investigate potential mechanism by which the derivative of digoxin selectively antagonizes RORγt transcriptional activity. Method Using molecular docking in combination with molecular electrostatic potential (MEP), we detected the interaction between the derivative of digoxin (Dhd) and ROR transcription factor (RORα,RORβ and RORγt), and the results were further confirmed by bioluminescent assay. Result Molecular docking demonstrated that Dhd could exclusively inhibit the conformation of RORγt;bioluminescent assay further indicated that RORγt was selectively antagonized by Dhd in a dose- and time-dependent manner. Conclusion Dhd can selectively suppress RORγt transcriptional activity.

Objetive Psoriasis is an autoimmune-related chronic inflammatory skin disease strongly associated with the dysfunction of Th17 cells. Retinoic acid-related orphan nuclear receptorγt (RORγt) plays a critical role in the differentiation and maturation of Th17 cells and in cell-derived immunologic derangement. We conducted this study to investigate potential mechanism by which the derivative of digoxin selectively antagonizes RORγt transcriptional activity. Method Using molecular docking in combination with molecular electrostatic potential (MEP), we detected the interaction between the derivative of digoxin (Dhd) and ROR transcription factor (RORα,RORβ and RORγt), and the results were further confirmed by bioluminescent assay. Result Molecular docking demonstrated that Dhd could exclusively inhibit the conformation of RORγt;bioluminescent assay further indicated that RORγt was selectively antagonized by Dhd in a dose- and time-dependent manner. Conclusion Dhd can selectively suppress RORγt transcriptional activity.

BACKGROUND: Repetitive brief and non-lethal cerebral ischemia can produce cumulative neuronal damage and vascular dementia; however, precisely injured patterns and mechanisms are still unclear. Thalamus is an important structure of learning and memory; meanwhile, it is also one of the selectively vulnerable regions of cerebral ischemia.However, there are a few reports about neuronal damage induced by repetitive cerebral ischemia.OBJECTIVE: To investigate the pathological damage and mechanism of neurons induced by repetitive cerebral ischemia in thalamus.DESIGN: Randomized controlled experimental study.SETTING: Department of Neurology, General Hospital of Chengdu Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out at the Animal Central Laboratory of the Third Military Medical University of Chinese PLA from March to December 1999. A total of 72 healthy male Wistar rats were randomly divided into sham operation group, single cerebral ischemic group,repetitive cerebral ischemic group, MK-801 treatment group and saline group.METHODS: Transient global cerebral ischemia models of rats were established with modified Pulsinelli-4 vessel occluing method for single 15-minute ischemia and repetitive three 5-minute ischemia at hourly intervals,followed by 5 hours, 2 days and 4 days of survival. Rats in sham operation group were not treated with burning vertebral artery and clipping common carotid artery. 45Ca autoradiography and light microscopy were used to determine the calcium accumulation and neuronal pathological changes of thalamus following repetitive cerebral ischemia as compared with single cerebral ischemia. The effects of MK-801, a N-methyl-D-aspartate (NMDA)receptor antagonist, were also examined.MAIN OUTCOME MEASURES: Distribution and degree of calcium accumulation and neuronal damage in the thalamus of rats in each group.RESULTS: Sham-operated rats revealed no abnormal calcium accumulation and neuronal damage in the thalamus. At 5 hours following ischemia,slightly abnormal calcium accumulation was found in the partial thalamus of the repeated ischemic group, and the neuronal damage was also relatively severer than that in the single ischemic group (0.98±0.19, 0.60±0.14, P＞ 0.05). At 2 days after ischemia, obviously abnormal calcium accumula tion and neuronal damage were shown in thalamus, and the degree of calcium accumulation and score of neuronal damage in repeated ischemic group were significantly severer than that in single ischemic group (1.62±0.31, 0.88±0.21, P ＜ 0.01). At 4 days, the thalamus calcium accumulation and neuronal damage were further increased, and also that in repeated ischemic group was significantly severer than that in single ischemic group (1.80±0.21, 1.02±0.23, P ＜ 0.01), especially marked calcium accumulation and cumulative damage were shown in the ventral thalamus. MK-801 significantly relieved the abnormal calcium accumulation and neuronal damage in the thalamus in repeated ischemic group, showing significant protection of thalamus neurons as compared with that in saline-treated group (0.20±0.12, 1.80±0.15, P ＜ 0.01).CONCLUSION: Repetitive non-lethal cerebral ischemia results in an intense cumulative damage in the ventral thalamus, and the excitatory amino acid and Ca2+ may play a major role in it.

Objective To explore the possible mechanism by which protein kinase C (PKC) involves in the neuronal apoptosis induced by cerebral ischemia/reperfusion Methods After the model of ischemia/reperfusion was established in male Wistar rats, PKC activity, FOS protein expression and neuronal apoptosis in their brains were observed The effect of PKC inhibitor, Dengzhanghua, on above indexes were studied at the same time Results Cerebral ischemia/reperfusion resulted in transloactional activation of PKC, accompanied with the increase of FOS expression and neuronal apoptosis Dengzhanghua prevented against the above changes Conclusion Activated PKC is involved in ischemia/reperfusion induced neuronal apoptosis by regulating FOS expression

AIM:To study pharmacokinetic of Pueraria Flavonid Nasal Drop in rabbits through nasal administration in comparison with oral administration. METHODS:Ten New Zealand rabbits were divided into two groups randomly and administrated nasally and orally (106.4 mg/kg of pueraria flavonid). HPLC was adopted to detect pueraria flavonoid content and DAS 2.0 was used to calculate bioavailability. RESULTS:The main pharmacokinetic parameters of nasal and oral administrations were AUC (0-∞)1 =(30.55?4.93)mg/(L?h),T max1 =(0.90?0.14)h,C max1 =(11.27?1.66)mg/L;AUC (0-∞)2 =(6.90?2.76)mg/(L?h),T max2 =(0.63?0.34)h,C max2 =(1.68?0.84)mg/L. Relative bioavailability of nasal delivery was 442.8%. CONCLUSION:Pueraria Flavonoid Nasal Drop is well absorbed in rabbits and has high bioavailability.