OBJECTIVE:To study the protective effects of Mongolian medicine Lomatogonium rotatum water extract against acute liver injury in mice caused by D-GlaN and CCl4. METHODS:60 mice were randomly divided into normal group(normal sa-line),D-GlaN model group(normal saline),positive control group(Kuihua hugan tablet,0.56 g/kg)and L. rotatum water extract high-dose,medium-dose and low-dose groups [3,1.5,0.75 g(crude drug)/kg] 10 rats in each group. They were given relevant medicine intragastrically once a day for consecutive 23 days. 30 min after last administration,those group were given D-GlaN ip to induce acute liver injury model except normal group. The activity of AST,ALT,ALP,and acetylcholinesterase(CHE)content in serum were detected,and liver index was calculated. Liver histopathology was observed and scored. Other 60 mice were selected, and then grouped,given medicine and detected in lab index with same method with above group,except for modeling method(ig, 0.1% CCl4). RESULTS:Compared with normal group,necrosis or degeneration of liver cells were obvious in 2 model groups, pathological score,the activity of AST,ALT and ALP increased while CHE content decreased;liver index of D-GlaN model group increased while that of CCl4 model group decreased (P<0.05). Compared with D-GlaN model group,the activity of AST,ALT and ALP and liver index decreased in L. rotatum water extract groups,while CHE content increased(P<0.05);pathological mor-phology had been improved,and pathological score decreased. Compared with CCl4 model group,ALT activity decreased in L. rota-tum water extract low-dose group,while CHE content and liver index increased;CHE content of L. rotatum water extract medi-um-dose group decreased,while liver index increased;the activity of AST,ALT and ALP decreased in L. rotatum water extract high-dose group,while CHE content increased(P<0.05). Pathological morphology of L. rotatum water extract groups had been im-proved to certain extent,and pathological score decreased. CONCLUSIONS:L. rotatum water extract could protect acute liver inju-ry in mice induced by D-GlaN and CCl4.

Objective To detect plasma amino-terminal pro-brain natriuretic peptide (NT-ProBNP) ,D-Dimer levels in the pa-tients with acute pulmonary embolism (APE) in order to investigate their change characteristics and significance .Methods Among 60 patients with suspected APE ,40 cases diagnosed by CT and meeting the thrombolytic condition were set as the experimental group and other 20 cases of excluded APE by CT as the control group .Plasma NT-ProBNP and D-dimer before treatment in the two groups and after 2-week thrombolytic therapy in the experimental thrombolysis were detected and compared .Results The diagnos-tic sensitivity of NT-ProBNP for early APE was 92 .3% and the specificity was 65% ,while the diagnostic sensitivity of D-dimer for early APE was 100% and the specificity was 70% ;the plasma NT-ProBNP and D-dimer levels before thrombolysis in the experi-mental group were significantly higher than those in the control group with statistical differences (P<0 .01) .Conclusion Plasma NT-ProBNP and D-dimer has important clinical significance for APE and can provide the basis for the early diagnosis and the cura-tive effect observation of the patients with APE .

Objective To investigate the effects of heat-killed Staphylococcus aureus (HKSA) on the apoptosis and expression of iNOS and IL-1β in THP-1 monocytes in the presence of high concentration of glucose.Methods THP-1 cells were cultured in medium containing 25.0 mmol/L(HG) or 5.5 mmol/L (LG,control) D-glucose for 12 h-8 d.The THP-1 cells cultured for 6 d were extracted on the 0-48 h with or without HKSA,then apoptosis and expression of iNOS and IL-1β were examined.Apoptosis was analyzed by flow cytometry and expressions of IL-1β and iNOS were quantitated by real-time PCR.Results The expression of iNOS and IL-1β in THP-1 monocytes was increased significantly in the presence of high concentration of glucose for 12-48 h(P＜0.05),reaching the highest level at 24 h and returned to baseline after 4 d.The expression was significantly lower than that of control after 4-6 d.Apoptosis rate was also increased significantly after 48 h to 4 days.HKSA infection enhanced apoptosis,but inhibited the expression of iNOS and IL-1 β in the presence of high concentration of glucose.The expression of iNOS and IL-1β increased significantly at 6 h(P＜0.01),reaching the highest level at 12 h,but the levels were significantly lower than those in control groups (P＜0.05).Conclusion These data suggest that high concentration of glucose can interfere with the anti-bacterial function of monocytes by reducing their expression of iNOS and IL-1β and enhancing their apoptosis.

Objective To study the determination method of total flavonoids in Gansu Astragali Radix and Hedysarum Polybotrys. Methods Calycosin glycosides etc. was selected as reference substances, comparison on the difference of absorption curves was done by ultraviolet spectroscopy and colorimetric method (NaNO2-Al(NO3)3-NaOH, AlCl3, Mg(Ac)2, NaOH, phosphomolybdic acid, HCl-Mg power). Results With colorimetric method, the maximum absorption wavelength of referrence and the test was inconsistent. The absorption peak shape was also different. With UV method, Calycosin glycosides in band Ⅱ (260 nm) showed a shoulder absorption. Astragali Radix and Hedysarum Polybotrys also showed characteristic shoulder absorptions in band Ⅱ with absorption wavelength at 263 nm and 265 nm. So the sample absorption wavelength is basically the same as that of the control sample. Conclusion Colorimetries usually used for determination of total flavonoids are not suitable for the comparison determination of Gansu Astragali Radix and Hedysarum Polybotrys. It is suitable for determining the contents of total flavonoids in samples by UV spectrophotometry at the band Ⅱ, which is the characteristic absorption band of isoflavone compound.

In order to investigate the characteristics of transdermal delivery of ferulic acid under the treated of microneedle arrays and the influence on permeability of rat skin capillaries, improved Franz-cells were used in the transdermal delivery experiment with the rat skin of abdominal wall and the length of microneedle arrays, different insertion forces, retention time were studied in the influence of characteristics of transdermal delivery of FA. The amount of FA was determined by HPLC system. Intravenous injection Evans blue and FA was added after microneedle arrays treated. Established inflammation model was built by daubing dimethylbenzene. The amount of Evans blue in the rat skin was read at 590 nm wavelength with a Multiskan Go microplate reader. Compared with passive diffusion group the skin pretreated with microneedle arrays had a remarkable enhancement of FA transport (P <0.01). The accumulation of FA increased with the enhancement of insertion force as to as the increase of retention time. Microneedle arrays with different length had a remarkable enhancement of FA transport, but was not related to the increase of the length. The research of FA on the reduce of permeability of rat skin capillaries indicated that the skin pretreated with microneedle arrays could reduce the content of Evans blue in the skins of rat significantly compared with the untreated group. The permeation rate of ferulic acid transdermal delivery had remarkable increase under the treated of microneedle arrays and the length of microneedle arrays ,the retention time so as to the insertion force were important to the transdermal delivery of ferulic acid.
Administration, Cutaneous ; Animals ; Coumaric Acids ; administration & dosage ; pharmacokinetics ; Male ; Needles ; Rats ; Rats, Sprague-Dawley ; Skin Absorption

This study investigated the protective effect of chrysin on hepatic fibrosis by regulating AMP-activated kinase (AMPK)-NOD-like receptor protein 3 (NLRP3) mediated pyroptosis pathway. The hepatic fibrosis model of mice was established by thioacetamide (TAA) in vivo. Except the control and chrysin alone groups, the mice were injected intraperitoneally with TAA at 100 mg·kg^-1, three times per week for the first week. From the 2^nd to 5^th week, mice were injected intraperitoneally with TAA at 200 mg·kg^-1, three times per week for the next 4 weeks. Chrysin groups were intragastrically administrated once per day to 5^th week. The histopathological changes were detected by HE and Masson staining. The levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assessed by the kits. All animal experiments were approved by the Medical Ethics Committee of Affiliated Zhongshan Hospital of Dalian University (DWLL2019060). LX-2 cells were stimulated by (transforming growth factor-β, TGF-β) in vitro. The protein expressions of AMPKα, p-AMPKα, NLRP3, cysteinyl aspartate specific proteinase-1 (caspase-1), gasdermin D (GSDMD) were detected by Western blot, and the mRNA levels of collagen-Ι, α-smooth muscle actin (α-SMA), interleukin-1β (IL-1β), IL-18, caspase-1, GSDMD were analysis by reverse transcription-polymerase chain reaction (RT-PCR). Chrysin attenuated the increases in serum AST and ALT levels in the TAA group, while significantly improved the changes of liver morphology, reduced liver tissue inflammatory cell infiltration and inhibited collagens deposition. Compared with TAA group, chrysin effectively activated AMPKα phosphorylation and inhibited hepatic NLRP3 inflammasome activation. Additionally, the protein expressions and mRNA levels of IL-1β, IL-18, caspase-1 and GSDMD in chrysin groups were decreased. Chrysin inhibited the expressions of collagen-Ι and α-SMA, enhanced the phosphorylation of AMPKα, and decreased the expressions of NLRP3 and GSDMD. Therefore, chrysin may inhibit inflammatory injury and pyroptosis possibly by activating AMPK and inhibiting NLRP3 inflammasome to alleviate hepatic fibrosis.

Objective To study the clinical significance of fecal smear examination on diagnosing intestinal flora imbalance in infantile diarrhea.Methods A sterile cotton swab was used to spread a layer of fresh feces quantum satis from a sterile container on a clean slide;the smear was fixed and stained with Gram′s methods after it was air-dried,then the specimen was observed with a microscopy(field lens 100 ? eye lens 10) and recorded.Results In the acute diarrhea group(40 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 6 cases(15%),≥50% Gram-positive cocci in 30 cases(75%) and ≥68% Gram-positive bacilli in 4 cases(10%).In the delayed and chronic diarrhea group(62 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 7 cases(11.29%),≥50% Gram-positive cocci in 44 cases(70.97%) and ≥68% Gram-positive bacilli in 6 cases(9.68%).In the normal control group(32 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 1 case(3.13%),≥50% Gram-positive cocci in 1 case(3.13%),and ≥68% Gram-positive bacilli in 17 cases(53.13%).For the distribution of 3 floras in the 3 groups,chi-squared test was performed,and the results showed that the difference was significant in Gram-positive cocci(?~2=47.76 P0.05).Conclusions Acute,delayed or chronic diarrhea can lead to flora imbalance.In order to timely and rapidly know the flora imbalance in children with diarrhea,clinically the simple,easily operated and practical smear staining method shall be widely applied.

The stability of melittin in different solvents (water, deoxygenated water, physiological saline, PBS, 50% ethanol, ethanol, glycerol)was studied and the results showed that the stability of melittin is not influenced by light, temperature and pH in 50% ethanol, which melittin can be completed dissolved when compared with ethanol and glycerol, in such, 50% ethanol was chosen as solvent storage when measured content of melittin. Then the effect of different concentrations of PBS, the pH of PBS and rat skin ho- mogenates were tested, and the results showed that melittin was degraded rapidly at low concentration solution and low ionic strength. Increasing pH of PBS and rat skin homogenate can accelerate the degradation of melittin. These researches provide an experimental ba- sis for further study of melittin.
Animals ; Drug Stability ; Ethanol ; chemistry ; Hydrogen-Ion Concentration ; Melitten ; chemistry ; Rats ; Skin ; drug effects ; Solvents ; chemistry ; Temperature

Objective To investigate the anti-apoptotic function of clusterin in LNCaP cell line and the role of clusterin antisense oligodcoxynucleotide(AS-ODN)in TNF-?-induced death of LNCaP cell. Methods The wild type LNCaP(group L),LNCaP transfected with the control vector(group M),LNCaP transfected with full-length clusterin expression vector(group A,ie,study group)were cultured.For the de- tection of cytotoxic effect of TNF-?,MTT and ELISA methods were used to determine the cell proliferation and apoptosis of the 3 clones,and the changes of proliferation and apoptosis in A cell after transfection of clusterin AS-ODN were also assessed.Results MTT method showed that the cell proliferation activity(A value)of groups L,M,and A were 0.84?0.03,0.85?0.04,0.95?0.03,respectively;the difference be- tween groups L and M was not significant(P＞0.05);but compared with group A the cell proliferation activ- ity was significantly lower in groups L and M(P＜0.01 for both).ELISA resuhs showed that the A values of groups L,M,and A were 0.59?0.04,0.62?0.03,0.33?0.04,respectively;the difference between groups L and M was not significant(P＞0.05);but compared with group A,the apoptosis rates were significantly higher in groups L and M(P＜0.01 for both).In group A,A values of cell proliferation activity in subgroups control,AS-ODN,TNF-?,TNF-?+AS-ODN were 1.30?0.03,1.25?0.03,0.99?0.03,0.80?0.03, respectively;the differences between each group were significant(P＜0.05 for all).And the A values of cell apoptosis in the above 4 groups were 0.02?0.00,0.21?0.02,0.63?0.07,1.16?0.04,respectively,the differences between each group were significant(P＜0.01 for all).Conclusions Stable transfection and subsequent expression of clusterin result in resistance to the cytotoxic effect of TNF-?.Transfection with clus- terin AS-ODN enhances cytotoxic effect of TNF-?in A cells.These results suggest that clusterin plays an im- portant role in anti-apoptotic function in LNCaP cell line.

Objective To evaluate the intervention effects of low-fluoride brick tea in the population, and to provide data for the prevention and control of the brick-tea type fluorosis. Methods Eighty-six Kazakh families with 5-12 years old children were selected and divided into two groups in the severe brick-tea type fluorosis areas of Akesai County of Gansu Province. Forty-six households were intervened by drinking low-fluoride brick tea as intervention group and another 40 households drank general brick tea as control group. The fluoride content in water, tea and urine was monitored and the total daily fluoride intake of adults and children was calculated by the fluoride content of the tea before and during intervention. The baseline prevalence of dental fluorosis was surveyed in all Kazakh school students aged 5 - 12 years before intervention, dental fluorosis prevalence were surveyed in two groups after the intervention. The fluoride content in water, urine,tea, and brick-tea samples was detected by iron electrode method, and dental fluorosis was diagnosed by Dean's method. Results The fluoride content of water were 0.36,0.50 mg/L respectively before and 42 months after intervention. The total daily fluoride intake of adults and children in the intervention group (being 4.39,5.12,5.38,4.49 mg in adults and 1.90,2.33 in children, 2.33, 1.94 mg for four calculations) were lower than those in control group (8.42,9.07,8.35,7.92 and 3.65,3.93, 3.62,3.43 mg). Except the second batch (530.4 mg/kg), the average fluoride content of the other 3 batches of low-fluoride brick tea(239.3,222.88,154.7 mg/kg) was lower than that of 4 batches of market brick tea(366.9,412.2, 286.0,379.6 mg/kg). The fluoride content of low-fluoride brick tea samples was in accordance with the national standard(< 300 mg/kg) in 16 of 21 samples in 4 the batches, and the qualifying rate was 76.19%(16/21). Only 5 of 21 market brick tea samples in 4 batches was qualified, accounting for 23.80%(5/21), both were significantly different(χ2= 11.52, P < 0.01). In 12, 36, 42 months after intervention, urine fluoride content in the intervention group of adult(1.84,1.23,1.77 mg/L) and children(1.55,0.65,1.10 mg/L) was less than that of the control group (adults: 3.37,3.68,3.02 mg/L, children: 2.64,1.64,2.62 mg/L), both being statistically significant (t value were 2.94,2.43,3.91,3.29,2.31,4.42, P < 0.01 or 0.05). The detective rate of dental fluorosis was 69.02%(127/184)at baseline among children. After the intervention, it lowered to [44.83% (13/29) in the intervention group, significantly lower than that in the control group[71.88%(23/32), χ2 = 4.60, P < 0.05]. Conclusion Low-fluoride brick tea can reduce the fluoride intake of the residents who drink brick tea, and alleviate excessive fluoride and the damage of high-fluoride.