[The analysis of long-term prognostic factors after laparoscopic liver resection for intrahepatic cholangiocarcinoma and establishment of survival Nomogram model].

Objective: To establish a survival prediction model based on the independent prognostic factors of long-term prognosis after laparoscopic liver resection(LLR) for intrahepatic cholangiocarcinoma(ICC). Methods: The clinical and pathological data of 351 consecutive patients with ICC who received radical LLR in 13 Chinese medical centers from August 2010 to May 2021 were collected retrospectively. There were 190 males and 161 females,aged(M(IQR)) 61(14)years(range:23 to 93 years). The total cohort was randomly divided into a training dataset(264 cases) and a validation dataset(87 cases). The patients were followed up by outpatient service or telephone,and the deadline for follow-up was October 2021. Based on the training dataset,the multivariate Cox proportional hazards regression model was used to screen the independent influencing factors of long-term prognosis to construct a Nomogram model. The Nomogram model's discrimination,calibration,and clinical benefit were evaluated through internal and external validation,and an assessment of the overall value of two groups was made through the use of a receiver operating characteristic(ROC) curve. Results: There was no significant difference in clinical and pathological characteristics and long-term survival results between the training and validation datasets(all P>0.05). The multivariate Cox analysis showed that CA19-9,CA125,conversion to laparotomy during laparoscopic surgery,and lymph node metastasis were independent prognostic factors for ICC patients after LLR(all P<0.05). The survival Nomogram was established based on the independent prognostic factors obtained from the above screening. The ROC curve showed that the area under the curve of 1, 3 and 5-year overall survival rates of patients in the training dataset were 0.794(95%CI:0.721 to 0.867),0.728(95%CI:0.618 to 0.839) and 0.799(95%CI:0.670 to 0.928),and those in the validation dataset were 0.787(95%CI:0.660 to 0.915),0.831(95%CI:0.678 to 0.983) and 0.810(95%CI:0.639 to 0.982). Internal and external validation proved that the model exhibited a certain discrimination,calibration,and clinical applicability. Conclusion: The survival Nomogram model based on the independent influencing factors of long-term prognosis after LLR for ICC(including CA19-9,CA125,conversion to laparotomy during laparoscopic surgery,and lymph node metastasis) exhibites a certain differentiation,calibration,and clinical practicability.

In Vitro and In Vivo Characterizations of Chiglitazar, a Newly Identified PPAR Pan-Agonist.

Solid rationales are still present for the identification of synthetic ligands to simultaneously target multiple PPAR subtypes for the treatment of T2DM. The purpose of this study was to characterize the in vitro and in vivo differential effects of chiglitazar, a non-TZD type of PPAR pan-agonist currently in phase III clinic development in China, from PPARγ-selective agonist like rosiglitazone. Chiglitazar showed transactivating activity in each PPARα, γ, and δ subtype and upregulated the expression of PPARα and/or PPARδ downstream genes involved in the key processes of lipid metabolism and thermogenesis. Comparable blood glucose lowering effect was observed between chiglitazar and rosiglitazone, but chiglitazar did not significantly increase the body weight in KKAy and fat pad weight in db/db mice. Chiglitazar had high distribution in liver, pancreas, and skeleton muscles but was less present in kidney, heart, and adipose in rats. Heart weight increase was not observed in rats treated with chiglitazar for 6 months at a dose as high as 45 mg kg(-1). The in vitro and in vivo differential features of chiglitazar are informative and encouraging for the further development of this synthetic ligand for the potential use in T2DM.

[An animal model for testing hypoglycemic and hypolipidemic drugs].

A new animal model of hyperglycemia and hyperlipidemia was established by treating normal Kunming mice with alloxan (iv). After different time intervals, the levels of blood glucose (BG) and serum triglyceride (TG) were determined. Forty-eight h after alloxan administration, the BG increased significantly, and the level of serum triglyceride reached maximum. The levels of blood glucose, serum triglyceride, cholesterol (CHOL) and free fatty acid (FFA) for vehicle treated mice were about 3.1, 4.0, 1.1, and 1.7 times those of normal Kunming mice, respectively. To evaluate the new animal model, four drugs were used. Two of them were antihyperglycemic drugs, metformin and a Recipe of Chinese Herbs (RCH). The other two were antihyperlipidemic agents, clofibrate and fenofibrate. All drugs showed positive effects on this kind of alloxan-diabetic Kunming mice. It can be concluded that this kind of alloxan-treated Kunming mice is useful for testing hypoglycemic and hypolipidemic drugs.

[Determination of berberine in biological specimen by high performance TLC and fluoro-densitometric method].

In this paper, a simple and specific high performance TLC and fluoro-densitometric method for separating and determining berberine in biological specimen was developed. Two hundred microliters of plasma or tissue homogenate mixed with 20 microliters of sodium lauryl sulfate (18%) and quinidine (300 ng, as internal standard) were extracted with 1 ml of chloroform. The developing solvent consisted of ethyl acetate-methyl acetate-methanol-water (5.4:4.6:1.2:1.0). The determination was carried out with a Shimadzu TLC-Scanner CS-910. Berberine spots were measured using an excitation wavelength of 350 nm and an emission wavelength of 550 nm, while using 350 nm and 450 nm for quinidine spots as the excitation and emission wavelength respectively. The average recoveries were 100.3% from plasma and 103.8% from tissue homogenate. The berberine levels in plasma and in tissues were compared in normal mice after oral administration of berberine (100 mg/kg) or the powder of Coptis chinensis (2 g/kg) containing the same amount of berberine. Simultaneously, the hypoglycemic effect of berberine and that of the powder of Coptis chinensis were measured in these mice. Results indicate that the concentrations of berberine in plasma and tissues given the powder of Coptis chinensis orally were higher than those given berberine and that the hypoglycemic effect of the powder was also stronger than that of berberine. Evidently, the changes of the blood glucose level and the level of berberine in plasma showed an opposite relationship.

Effects of glutathione depletion using buthionine sulphoximine on the cytotoxicity in mammalian cells and human tumor cells in vitro.

An inhibitor of glutathione biosynthesis, buthionine sulphoximine (BSO), was used to deplete the endogenous thiols in mammalian cells in vitro. In this study, the cytotoxicity of BSO and BSO combined with the hypoxic cell radiosensitizer misonidazole (MISO) was investigated. Both aerobic and hypoxic cytotoxicity of MISO was found to be increased. The concentration of BSO required to reduce the colony forming ability to 50% (Cc) for the chronic cytotoxicity on V79 cells was 0.03 mmol/L under aerobic condition, while the Cc for the acute cytotoxicity on V79 cells under hypoxic and aerobic conditions was 0.4 and 0.5 mmol/L. The growth inhibition rate of human tumor cells K562 and SGC-7901 by BSO was 6.89-26.06% and 12.01-55.69%, respectively. Enhanced cytotoxicity activity was observed when BSO was used in combination with cis-dichlorodiamino Pt(II) or 5-fluorouracil.

Enhancement of systemic delivery of met-enkephalin and leu-enkephalin eyedrops with permeation enhancers.

It was found that methionine enkephalin (Met-Enk) and leucine-enkephalin (Leu-Enk) can be delivered efficiently into the systemic circulation through the outer route in rabbits. When 50 mcl of 1% Met-Enk eyedrops without absorption enhancers were instilled into eyes, the rabbits' blood concentration rose from 92 pg/ml to 153 pg/ml in 10 min. When either of two permeation enhancers (BL-9 or Brij-78) was added at a 0.5% concentration to the ophthalmic solution, the systemic absorption of 1% Met-Enk was markedly improved. Its blood level was increased to approximately 3.4 times that without the enhancer. When the eyedrops of 0.3% Met-Enk plus 0.5% absorption enhancer were administered, the blood concentration reached higher than that reached by 1% Met-Enk without permeation enhancers. Similar results were obtained with Leu-Enk except the systemic absorption was enhanced by BL-9 and Brij-78 even further to 10-fold and 8.3-fold, respectively. These results indicate that the systemic delivery of Met-Enk and Leu-Enk through eyes, especially with the addition of permeation enhancers, is a feasible alternative route to parenteral injection.

Effects of permeation enhancers BL-9 and Brij-78 on absorption of four peptide eyedrops in rabbits.

Systemic absorption of 4 peptide drugs through ocular route was enhanced by permeation enhancers, BL-9 and Brij-78. Gonadorelin (LHRH) was the smallest molecule (M(r) = 1200) and its systemic delivery was most efficiently enhanced by BL-9 (17.0-20.8 times) and Brij-78 (13.9-21.5 times). Although bombesin (M(r) = 1620), atrial natriuretic peptides (ANP) (M(r) = 3240), and adrenocorticotropic hormone (ACTH) (M(r) = 4540) had molecular weight ranged widely, their systemic absorption through ocular route was enhanced by BL-9 (6.1-8.3 times) and Brij-78 (6.5-9.0 times) in about the same degree. BL-9 enhanced systemic absorption of peptide drugs faster and reached peak peptide concentrations in 5-20 min. On the other hand, Brij-78 took 20-60 min to reach to peak concentration of peptide drugs in the blood. These results indicate that systemic delivery of peptide drug through ocular route is a feasible one particularly when the absorption enhancers are used.

Effects of some natural products on sugar cataract studied with nuclear magnetic resonance spectroscopy.

Four commercial anticataract drugs and five flavonoids isolated from Chinese herbs were studied on their effects to inhibit sugar cataract formation in vitro using 13C-nuclear magnetic resonance spectroscopy (MRS). Pa-Wei-Di-Huan-Wan (PWDHW) and Zhang-Yan-Ming (ZYM) were effective in inhibiting sorbitol formation, whereas Bai-Nei-Ting (BNT) and Pearl Min-Mu-Ye (Pearl MMY) were ineffective in inhibiting sorbitol formation. Among five flavonoids tested, four were moderately effective in inhibiting sorbitol formation, and one was totally ineffective up to 1 x 10(-4)M used.

[Re-estimation on the role of artificial pneumoperitoneum in the treatment of pulmonary tuberculosis haemoptysis].

The response of 150 patients with pulmonary tuberculosis haemoptysis treated by artificial pneumoperitoneum, when they failed to the regular hemostatic drugs, is analysed. The data showed that 101 cases (67.3%), got favorable results, and there was less effective (24.3%) in 35 patients; only 14 cases (9.3%) failed. Thus the total response rate was 90.6%. The conclusion is that artificial pneumoperitoneum is a rather safe, simple economic and effective method in treatment for hemoptysis of pulmonary to tuberculosis.

Ocular and cardiovascular pharmacology of tetramethylpyrazine isolated from Ligusticum wallichii Franch.

Tetramethylpyrazine (TMP), the active principle in Ligusticum wallichii, D-timolol, and L-timolol were compared for their effects on retinal and choroidal blood flow (RCBF), blood pressure (BP), and heart rate (HR). TMP (10 mg.kg-1) increased RCBF by 44% and did not affect systemic BP or HR. D-Timolol (4 mg.kg-1 iv) had a tendency to increase RCBF but did not affect systemic BP and HR either. L-Timolol (0.4 mg.kg-1 iv), on the other hand, decreased RCBF by 18% and reduced both systemic BP and HR. In isolated preparations, TMP increased coronary artery blood flow with slight vasodilation, but vasoconstriction in renal, femoral and mesenteric arteries. These results indicate that TMP could be used to prevent or alleviate certain ischemic retinal degenerations without producing significant cardiovascular side effects. The in vitro vasoconstriction actions of TMP (0.2 mg.ml-1) were blocked by propranolol and phenoxybenzamine, indicating that adrenergic mechanism might be involved in TMP action.

Systemic absorption of oxytocin and vasopressin through eyes in rabbits.

In order to avoid injections, oxytocin and vasopressin were systemically administered through ocular route. Although oxytocin and vasopressin are small peptide drugs with only 9 amino acids, their absorptions were relatively poor. The systemic absorption of these peptides through eyes was markedly enhanced by the addition of an absorption enhancer, 0.5% Brij-78 (polyoxyethylene-20-stearyl ether), in the eyedrops of these peptides. It is concluded that it is feasible to administer oxytocin and vasopressin as eyedrops to avoid parenteral injections.

Effects of D-timolol and L-timolol on ocular blood flow and intraocular pressure.

Effects of D-timolol and L-timolol on IOP were compared with two rabbit models. When the drug solution was injected into vortex vein, 1% D-timolol produced ocular hypotension just like 0.5% L-timolol except D-timolol was less potent than L-timolol to lower the IOP. On the other hand, when 0.5% of D-timolol and L-timolol were instilled into the rabbit eye on IOP recovery model both agents showed equipotency to delayed the IOP recovery. Effects of D-timolol and L-timolol on ocular blood flow were also studied with two rabbit models. D-Timolol at 0.5% did not affect the ocular pulsatile blood flow measured with Langham's pneumatonometer whereas 0.5% L-timolol significantly suppressed it. D-Timolol (0.5%) was found to increase retinal and choroidal blood flows measured with laser Doppler method whereas L-timolol suppressed it. These results indicate that D-timolol though less potent than L-timolol to lower IOP, is superior over L-timolol to improve the blood flow in retina and choroid.

Adjustment of blood sugar levels with insulin and glucagon eyedrops in normal and diabetic rabbits.

The ophthalmic solutions of insulin and glucagon plus permeation enhancers, BL-9 and Brij-78, were administered to rabbit eyes to study the systemic absorption of these peptide drugs and their effects to affect the blood glucose levels. When plain insulin eyedrops were given, the blood sugar levels were little affected. When insulin plus permeation enhancers were instilled to the eyes, the blood glucose reduced markedly in both normal and diabetic animals. Although sufficient concentrations of glucagon can be reached with plain glucagon eyedrops, the addition of permeation enhancers increased the systemic absorption of glucagon through eyes markedly. As a result, the blood sugar levels were raised markedly by glucagon eyedrops at much lower concentrations. These results indicate that systemic delivery of insulin, glucagon, and other peptide drugs through eyes is a feasible alternative route to parenteral injection particularly when permeation enhancers are added.

[The hypoglycemic effect of clausenacoumarine].

Clausenacoumarine is a compound isolated from the leaves of Clausena lansium (Lour.) Skeels which grows widely in South China. The studies show that clausenacoumarine can lower blood glucose level in normal mice and alloxan diabetic mice at 200 mg/kg.d for three days orally, and antagonize the elevation of blood glucose caused by injecting adrenaline in normal mice. No effect on blood lactic acid was observed.