Objective:To compare the anti-thrombin activity and the effects on the coagulation pathway between Whitmania pigra Whitman and Hirudinaria manillensis to provide scientific reference for the anticoagulation mechanism revelation for Hirudo. Methods:Anti-thrombin titration and chromogenic substrate assay-extraction-HPLC were applied to study the anti-thrombin activity of Whitmania pigra Whitman and Hirudinaria manillensis. APTT, PT and TT were determined by a clotting assay to compare the effects on the path-way of blood clotting. Results:The anticoagulation activity order measured by anti-thrombin titration was living Hirudinaria manillensis> dried Hirudinaria manillensis > > dried Whitmania pigra Whitman. The results of chromogenic substrate assay-extraction-HPLC in-dicated that the low dose of aqueous extract promoted the thrombin activity, while the high dose inhibited the thrombin activity. Hirudi-naria manillensis significantly inhibited the activity of thrombin, while Whitmania pigra Whitman showed weak anti-thrombin activity only at the higher dose. All leeches could prolong APTT, PT and TT. However, living Hirudinaria manillensis mainly affected TT, and dried Hirudinaria manillensis mainly affected APTT. Dried Whitmania pigra Whitman dramatically influenced APTT and TT. All the results indicated that the anticoagulant activity of Whitmania pigra Whitmanis was significantly higher than that of Hirudinaria manillen-sis. Conclusion:There are notable differences in the anti-thrombin activity and the effect on the pathway of blood clotting between Whitmania pigra Whitman and Hirudinaria manillensis.

Objective To establish a pre-column derivation HPLC-UV method for the content determination of hyodeoxycholic acid in artificial calculus bovis. Methods The hyodeoxycholic acid was derived by 2-bromo-2’-acetonaphthoneat using triethylamine as the catalyst in 60 ℃ water bath. After that, a HPLC-UV method was established to determine the content of hyodeoxycholic acid in artificial calculus bovis. Results When the derivatising time at 60 ℃ water bath was 50 min, the radio of the molar amount of derived reagents and hyodeoxycholic was over 20:1 and the radio of catalyst and hyodeoxycholic was over 15:1; the reaction was completed. The calibration curve was linear within the range of 0.1–2 μg for hyodeoxycholic acid (r=0.999 7), and the average recovery was 97.85% (RSD=1.6%). In this sample, the content of hyodeoxycholic is 4.12%. Conclusion The method is with high sensitivity, highly reproducible, reliable and accurate for the content determination of hyodeoxycholic acid in artificial calculus bovis.

Objective:To investigate the efficacy of levodopa and benserazide hydrochloride combined with pramipexole in the treatment of Parkinson's disease in 30 patients and their effects on neurotransmitters and oxidative stress response.Methods:A total of 90 patients with Parkinson's disease admitted to Jinhua People's Hospital from January 2020 to February 2022 were included in this study. They were randomly assigned to undergo treatment with levodopa and benserazide hydrochloride (levodopa and benserazide hydrochloride group), pramipexole (pramipexole group), or their combination (combined therapy group), with 30 patients in each group. All patients were treated for 12 consecutive weeks. Clinical efficacy, levels of brain neurotransmitters (dopamine, 5-hydroxytryptamine, norepinephrine, and substance P), and oxidative stress response (superoxide dismutase, malondialdehyde, homocysteine levels) were compared among the three groups.Results:Total response rate in the combined therapy group was 96.67% (29/30), which was significantly higher than 66.67% (20/30) in the levodopa and benserazide hydrochloride group and 76.67% (23/30) in the pramipexole group ( χ2 = 8.65, P < 0.05). After treatment, dopamine, 5-hydroxytryptamine, norepinephrine, substance P, superoxide dismutase, malondialdehyde, and homocysteine levels in the combined therapy group were (9.05 ± 1.24) ng/mg, (89.49 ± 10.69) μg/L, (15.16 ± 1.36) ng/mg, (102.8 ± 15.36) μg/L, (88.40 ± 10.04) kU/L, (5.5 ± 2.31) μmol/L, and (9.20 ± 3.36) μmol/L, respectively, which were superior to (6.61 ± 1.02) ng/mg, (68.52 ± 9.52) μg/L, (12.33 ± 1.24) ng/mg, (151.64 ± 16.03) μg/L, (74.99 ± 7.28) kU/L, (9.27 ± 3.07) μmol/L, and (13.52 ± 3.64) μmol/L in the levodopa and benserazide hydrochloride group and (7.22 ± 1.09) ng/mg, (79.52 ± 10.20) μg/L, (13.92 ± 1.31) ng/mg, (131.30 ± 15.65) μg/L, (80.59 ± 8.24) kU/L, (7.53 ± 2.93) μmol/L, (11.35 ± 3.71) μmol/L in the pramipexole group ( F = 38.53, 32.05, 35.49, -73.42, 18.42, -22.65, -12.13, all P < 0.05). Conclusion:Levodopa and benserazide hydrochloride combined with pramipexole are highly effective on Parkinson's disease. The combined therapy can effectively improve brain neurotransmitters and regulate oxidative stress response.

Objective:To investigate the correlation between serum ceruloplasmin level and elevated impulsivity in elderly patients with Parkinson′s disease (PD).Methods:227 elderly PD patients treated in Jinhua People′s Hospital from October 2019 to June 2021 were selected as the research objects. They were grouped according to the 75th percentile of serum ceruloplasmin. 0-75th percentile was defined as normal and >75th percentile was defined as high level. The differences of second-order and first-order factor scores of Barratt′s Impulsivity Scale Version 11 (BIS-11) between the two groups were observed. After balancing the general characteristics, the third part of Parkinson′s Disease Rating Scale (MDS-UPDRS), Hoehn&Yahr Scale, Addenbrooke Cognitive Examination Revised Edition (ACE-R), clinical treatment plan and other data, the correlation between ceruloplasmin and BIS-11 was observed.Results:According to the 75th percentile level of ceruloplasmin, 56 patients were included in the high-level group and 171 patients were included in the normal group. The level of ceruloplasmin, the ratio of female patients, MDS-UPDRS Ⅲ, Hoehn&Yahr Scale and ACE-R score in the high-level group were significantly higher than those in the normal group (all P<0.05). The second-order unplanned and corresponding first-order self-control and first-order self-knowledge complexity in the high-level group were higher than those in the normal group (all P<0.05). There was no significant difference in second-order attention, first-order attention, first-order cognitive instability, second-order motivation, first-order motivation, first-order stability and BIS-11 score between the two groups (all P>0.05). After removing the confounding factors by multifactor logistic analysis, ceruloplasmin was correlated with second-order unplanned and its corresponding first-order factors (self-control and self-knowledge complexity) (all P<0.05). Conclusions:The serum ceruloplasmin level in elderly PD patients is related to the elevated level of BIS-11 unplanned impulse.

Objective:To investigate and compare the effects of water extracts of Whitmania pigra Whitman and Hirudinaria ma-nillensis Lesson on the angiogenesis of Tg (kdrl:mCherry) zebrafish. Methods:The zebrafish embryos 6-8 hours after fertilization (6-8hpf) were transferred to the culture medium containing Whitmania pigra Whitman or Hirudinaria manillensis Lesson extract at different concentrations, and the culture medium containing the drugs was replaced every 24 h. And then, at 72 hpf, the larvalmorphology and intersegmental vessels were observed under a microscope. The hatchability of 48-and 72-hpf embryos, and the number of intersegmen-tal vessels and the heart rate of 72-hpf juveniles were measured. Results:Compared with the control group, when the concentration of Whitmania pigra Whitmanis was higher than 30μg· ml-1 , and the concentration of Hirudinaria manillensis Lesson was higher than 20μg· ml-1, the number of intersegmental vessels was significantly reduced (P<0. 01). Compared with the control group, at 48 hpf, when the concentration of the drug groups was higher than 40 μg· ml-1 , the hatchability of the two groups significantly decreased ( P<0. 01);at 72 hpf, the hatchability of Whitmania pigra Whitman decreased significantly at the concentration of 100 μg·ml-1 (P<0.01), while the hatchability of Hirudinaria manillensis Lesson decreased significantly at the concentration of 80 μg· ml-1(P <0. 01). There was no obvious yolk sac edema, pericardial edema and spine curvature in the two groups. The heart rate decreased sig-nificantly (P<0. 01), while was still within the normal range. Conclusion:Both Whitmania pigra Whitman and Hirudinaria manillen-sis Lesson have notable anti-angiogenic activity, and the anti-angiogenesis activity of Hirudinaria manillensis Lesson is stronger. They both have effects on the development of zebrafish embryos, while the toxicity is not obvious.