Objective To investigate the expressions of T helper cell 1 (Th1)-associated chemokine receptors CXCR3, CCR5 and T helper cell 2 (Th2)-associated chemokine receptor CCR3 in the spleen tissues of rats with cirrhosis and hypersplenism and probe into the balance between Th1/Th2 lymphocyte subsets.Methods Experimental study was adopted.Forty-six male SD rats were randomized into the hypersplenic group (n =36) and the control group (n =10).In the hypersplenic group, the rats were fed with 40％ CCl4 peanut oil solution (3.0 mL/kg, twice per week) and 15％ white spirit for 8 weeks to build the hypersplenic model.The rats in the control group received normal feeding.The animal models with cirrhosis and hypersplenism were confirmed by liver function test, routine blood test, HE staining and Masson staining after visual inspection.The expressions of chemokine receptors of CXCR3, CCR5 and CCR3 were detected by immunohistochemical staining and Western blot.Measurement data with normal distribution were presented as x ± s.Comparison between groups was done using the independent sample t test.Results Results of visual inspection: the rats in the hypersplenic group suffered from severe hair-shedding, metal fatigue and inappetence, with hair dimming and inactivity.There were rats dead successively 5 weeks after model establishment and 19 rats finally survived.The rats in the control group had color and gloss hair, with good appetite and spirits.They were active and sensitive to external stimulation.Changes of pathological morphology in liver: in the hypersplenic group, the fibers became denser and disordered, making normal structure of liver tissues destroyed.The hepatic lobules separated by fibrous bundle and proliferative hepatic cell mass were segmented and surrounded by thick fibrous,leading to the formation of pseudolobule.Disorganized hepatocytes suffused adipocytes, the nucleus of heterocysts enlarged or even multinucleated cells appeared.There was no change in the control group.Changes of pathological morphology in spleen: the rats in the hypersplenic group had slightly swelling spleen with the areas of red pulp increased and whit pulp disappeared gradually.Vascular endothelium became thicker and proliferated.Thickened central artery and fibrosis were depicted.Splenic sinusoid extended.There was no change of spleen tissues in the control group.Changes of liver function : the levels of ALT and AST of rats were (264 ± 111) U/L and (687 ± 299) U/L in the hypersplenic group,which were significantly different from (27 ± 8)U/L and (124 ± 20)U/L in the control group (t =5.64, 4.98,P ＜ 0.05).The level of total protein was (54 ± 8)g/L in the hypersplenic group, which was significantly different from (65 ± 3)g/L in the control group (t =-3.35, P ＜ 0.05).Changes of peripheral blood cell count: the white blood cell (WBC) count in the hypersplenic group was (23.9 ± 5.0) × 109/L, which was significantly different from (6.2 ± 2.4) × 109/L in the control group (t =3.50, P ＜ 0.05).The red blood cell count and platelet count in the hypersplenic group were (6.3 ±0.7) × 1012/L and (418 ± 124) × 109/L, which were significantly different from (8.0 ± 0.6) × 1012/L and (1 109 ± 161) × 109/L in the control group (t =-2.28,-4.92, P ＜ 0.05).Results of immunohistochemical staining: the cytomembrane and/or cytoplasm stained yellow, brown or sepia were defined as positive performance of CXCR3, CCR5 and CCR3.The absorbance A values of CXCR3 and CCR5 were (81.7 ±24.4) × 10-3 and (3.6 ± 1.3) × 10-3 in the hypersplenic group, which were significantly different from (19.2 ± 5.8) × 10-3 and (1.2 ± 0.4) × 10-3 in the control group (t =16.22, 9.09, P ＜ 0.05).The absorbance A value of CCR3 was (8.8 ±3.7) × 10-3 in the hypersplenic group and (7.9 ±2.8) × 10-3 in the control group, respectively, showing no significant difference between the 2 groups (t =0.87, P ＞ 0.05).The rates of positive cells of CXCR3 and CCR5 was 52％ ± 9％ and 19％ ± 5％ in the hypersplenic group, which were significantly different from 21％±5％ and 10％±3％ in the control group (t =17.31, 8.21, P ＜0.05).The rates of positive cells of CCR3 in the hypersplenic group and control group were 35％ ± 9％ and 33％ ± 14％, respectively, showing no significant difference between the 2 groups (t =0.43, P ＞ 0.05).Results of Western blot test : the relative expressions of CXCR3 and CCR5 were 2.45 ± 0.85 and 0.94 ± 0.48 in the hypersplenic group, which were significantly different from 1.31 ± 0.95 and 0.32 ± 0.26 in the control group (t =2.62, 2.91, P ＜ 0.05).The relative expression of CCR3 was 0.47 ± 0.27 in the hypersplenic group, which was significantly different from 0.92 ± 0.67 in the control group (t =-2.18, P ＜ 0.05).Conclusion The abnormal expression of chemokine receptors in the spleen tissues of rats with cirrhosis and hypersplenism induced by CCl4 suggests that functional imbalance of Th1/Th2 lymphocyte subsets may play an important role in the regulation of peripheral blood cytopenia.

Objective To investigate the intra-splenic blood cell count of posthepatitic cirrhotic portal hypertension,and compare it with patients' peripheral blood cell count to explore the role the spleen plays in peripheral cytopenia often seen in posthepatitic cirrhotic portal hypertension.Methods A prospective study was made on 15 cases with post hepatitis B cirrhotic portal hypertension undergoing splenectomy.Intrasplenic blood was sampled from upper pole,hilus (central pole),and lower pole of the spleen respectively for blood cell count.Results were compared with that of preoperative peripheral blood.Results There were significant statistical differences in the WBC count between splenic blood and peripheral blood,(11.20 ± 4.73) × 109/L vs.(4.06 ± 1.75) × 109/L,t =5.05,P ＜ 0.05),and in PLT count,(182.45±66.57) × 109/L vs.(63.54 ±28.40) × 109/L,t =7.285,P ＜0.05.There was no differences in the RBC count,(3.55 ± 0.94) × 1012/L vs.(3.01 ± 0.62) × 1012/L,t =1.874,P ＞ 0.05.Positive correlations were found between splenic PLT count and peripheral PLT count (r =0.610,P ＜0.05).Conclusions In posthepatitic B cirrhotic portal hypertension patients the intra-megalosplenic PLT and WBC count are significantly higher than that in peripheral blood.Megalosplenic PLT count correlates positively with peripheral PLT count.

Objective:To observe the effects of different moxibustion time on cartilage morphology,tumor necrosis factor(TNF)-α and interleukin(IL)-10 of the knee joint in rats with knee osteoarthritis(KOA),and to explore the best treatment time of moxibustion for KOA.Methods:Healthy male Wistar rats were randomly divided into a blank group,a model group,a 15-minute-moxibustion group,a 30-minute-moxibustion group,and a 60-minute-moxibustion group,with 10 rats in each group.Except for the blank group,the KOA model was established in all groups by injecting sodium iodoacetate solution into the knee joint cavity of rats.Rats in the 15-minute-moxibustion group,the 30-minute-moxibustion group,and the 60-minute-moxibustion group were all treated with mild moxibustion intervention for 15 min,30 min,and 60 min,respectively at Neixiyan(EX-LE4)and Dubi(ST35)points near the patella,3 times a week for 4 weeks,12 times in total.Rats in the blank group and the model group were fixed for 30 min without moxibustion intervention.Macroscopic observation for the smoothness of knee cartilage surface was performed after the intervention.Hematoxylin-eosin staining,toluidine blue staining,and Mankin score were used to evaluate the pathological changes in the cartilage.The expression levels of TNF-α and IL-10 in the serum were detected by enzyme-linked immunosorbent assay.Results:Compared with the blank group,the articular cartilage surface in the model group was rough,the chondrocyte arrangement was irregular,the Mankin score and the serum TNF-α expression were significantly increased(P＜0.05),while the expression of serum IL-10 was significantly decreased(P＜0.05).Compared with the model group,the articular cartilage surface was smoother,the chondrocytes were arranged neatly,the Mankin score and serum TNF-α expression level were significantly lower in the three moxibustion intervention groups(P＜0.05);the serum IL-10 level in the 30-minute-moxibustion group and the 60-minute-moxibustion group was increased significantly(P＜0.05).Compared with the 15-minute-moxibustion group,the articular cartilage surface in the 30-minute-moxibustion group and the 60-minute-moxibustion group was smoother,the chondrocyte arrangement was more regular,the Mankin score and the serum TNF-α level were decreased significantly(P＜0.05),and the serum IL-10 level was increased(P＜0.05).There was no significant difference in the serum TNF-α or IL-10 level between the 30-minute-moxibustion group and the 60-minute-moxibustion group(P＞0.05).Conclusion:Moxibustion can obviously improve the morphology and structure of KOA articular cartilage,protect articular cartilage,inhibit cartilage inflammation,and delay KOA cartilage degeneration.Moxibustion's effect is closely related to moxibustion time;the therapeutic effect of the 30-minute-moxibustion and the 60-minute-moxibustion is better than that of the 15-minute-moxibustion.

OBJECTIVETo make a qualitative analysis on chemical components in Ligustrum lucidum by UPLC-ESI-Q-TOF-MS.

METHODACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 microm) column was adopted, with methyl cyanides-0.1% formic acid as the mobile phase for gradient elution; ESI ion source was used for mass spectra, and data were collected in positive and negative mode.

RESULTFourteen compounds of L. lucidum were identified by analyzing positive and negative ion mass spectra information and element composition and comparing controls with data from relevant literature.

CONCLUSIONAfter the separation by ultra high performance liquid chromatography, relative molecular mass was determined by mass spectra, and chemical compounds in L. lucidum were determined by information search through relevant literature data, in order to provide powerful therapeutic material basis for L. lucidum.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ligustrum ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; methods

Objective To investigate the causes of peripheral cytopenia in patients with posthepatitic cirrhosis and portal hypertensive splenomegaly.Methods The clinical data of 183 patients with hepatitic cirrhosis and portal hypertensive splenomegaly complicated by peripheral cytopenia who were operated in our hospital in the past 17 years were retrospectively studied.Results All these patients underwent splenectomy.Before operation,all these patients had one or more types of peripheral cytopenia (cumulative cytopenia:390 patient-times).After splenectomy,blood counts in 79.2％ returned to normal;in 15.9％ increased but failed to reach normal levels;and in 4.9％ became lower than before operation.5 patients died soon after operation.Conclusion Hypersplenism is the main cause for the peripheral cytopenia most cirrhotic portal hypertension patients.Splenectormy is an effective method to treat hypersplenism.

Objective To investigate the genetic carrier rate of thalassemia and its gene mutation types as well as the distribution characteristics among couples of reproductive age in Dongfang City of Hainan Province,and to provide a basis for making prevention and control strategies against thalassemia.Methods Samples were collected from 1 000 couples undergoing premarital and pregestational screenings for thalassemia in Dongfang City of Hainan Province from September 2012 to March 2013,in which the positive ones in preliminary screening were further tested by genetic diagnoses and the genotypes were retrospectively analyzed.Results Among 1 000 couples,322 spouses were diagnosed with thalassemia gene mutation and the carrying rate was 16.10％ (322/2 000).In those carriers,246 spouses were α-thalassemia and the carrying rate was 12.30％ (246/2 000),accounting for 76.40％(246/322) of all thalassemia carriers,among them,there were 197 cases of α-deficiency genotype,accounting for 61.18％ (197/322),32 carried mutated α-gene,accounting for 9.94％ (32/322),17 carried both deleted and mutated α-gene,accounting for 5.28％ (17/322);43 spouse were β-thalassemia and the carrying rate was 2.15％(43/2 000);33 spouse were both α-and β-thalassemia and the carrying rate was 1.65％ (33/2 000).In spouses diagnosed with α-thalassemia,the major genotype was-α37/αα,accounting for 19.25％ (62/322);the second ranked was-α4.2/αα,accounting for 17.70％ (57/322),and the third ranked was--SEA/αα,accounting for 8.70％ (28/322).In spouses diagnosed with β-thalassemia,the major genotype was CD41-42/N,accounting for 9.63％ (31/322).Conclusions The population carrying rate of thalassemia in Dongfang City of Hainan Province is high,and its major type is α-thalassemia.For the purpose of decreasing the birth rate of thalassemia,major,local public health department should attach great importance to thalassemia prevention,and strengthen premarital and pregestational screening for thalassemia.