To optimize the water extraction process of modified Tabusen-2 by response surface methodology (Box-Behnken) and investigate its in vitro efficacy.

Taking the mass fractions of caffeic acid and geniposide as the comprehensive score, response surface methodology was used to optimize the soaking time, material-to-liquid ratio, extraction time and extraction times. The RAW 264.7 cells induced by nuclear factor-κB receptor activator ligand (RANKL) were co-cultured with the optimal extracts for 5 d, and then treated with tartrate-resistant acid phosphatase (TRAP). The TRAP-positive cells were stained and observed under a microscope to evaluate the in vitro efficacy of the extract.

The extraction time, soaking time, and material-to-liquid ratio were selected as the inspection factors, and the comprehensive score of caffeic acid and geniposide content was used as the evaluation index. After optimization by response surface method, the optimal extraction condition was 540 min; the liquid ratio was 1∶12 (g·mL^-1); the extraction time was 44 min, and the extraction times were two times. The extract obtained under this condition had a significant inhibitory effect on RAW264.7 cells induced by RANKL, could reduce TRAP-positive cells, and inhibited multinucleation.

The water extraction process of modified Tabusen-2 optimized by response surface analysis method is reproducible, stable and feasible, and has good in vitro pharmacodynamic activity, laying a reliable foundation for future in vivo pharmacodynamic experimental research.

To explore the effect and mechanism of compound cortex phellodendri fluid in bacterial vaginosis rats via TLR4/NF-κB pathway.

Fifty-five SD female rats were selected to construct bacterial vaginosis model by injection of mixed bacterial liquid. After successful modeling, the rats were treated with compound cortex phellodendri fluid, metronidazole vaginal gel, and Honghe fujie lotion, respectively. After 7 d of continuous treatment, inflammatory cell infiltration and glycogen accumulation in rat vaginal tissues were observed by HE staining. ELISA method was used to detect the expression level of IL-1β, IL-2, IL-10, IL-6, and TNF-α in the vaginal lavage. The distribution of NF-κB p65 in rat vaginal tissues were located by immunohistochemical method. And the protein expression level of TLR4 and NF-κB p65 were detected by Western blot.

Compared with those of the model group, the infiltration of inflammatory cells in compound cortex phellodendri fluid group was reduced, and the accumulation of glycogen was significantly increased (P<0.01), the concentrations of IL-1β (P<0.01), IL-6 (P<0.05), IL-2, and TNF-α in the vaginal lavage were decreased, while the IL-10 concentration increased (P>0.05), the protein expression level of TLR4 and NF-κB p65 decreased (P<0.05), and the NF-κB p65's translocated nuclear were suppressed (P<0.01).

Compound cortex phellodendri fluid has a definite therapeutic effect on bacterial vaginosis rats. Its mechanism may be related to the inhibition of TLR4/NF-κB pathway and adjustion of immune imbalance on Th1/Th2.

To understand the innovation points of drugs marketed after the implementation of the U.S. breakthrough therapy designation (BTD) program, and provide a reference basis for the development of innovative drugs and the review of breakthrough therapy drugs in China.

As of 2021, all BTD drugs approved by the U.S. FDA for marketing were collected, and the characteristics of these drugs in terms of innovative therapeutic modalities, treatment mechanisms, and technological breakthroughs were categorized and statistically analyzed.

The US FDA received 1 192 BTD drug applications and approved 242 for marketing. Among the marketed drugs, 42 (17%) were new drugs to fill clinical treatment gaps, 99 (41%) were drugs for expanded indications and new applicable populations, 27 (21%) were drug combinations for combination therapy, 16 (7%) were new drugs with more clinical treatment advantages, 31 (12.8%) were drugs with new mechanisms of action, and 12 (5%) were drugs prepared in new dosage forms or processes.

The substantial improvement of the clinical efficacy of BTD drugs is reflected in the innovation of drugs and advancement of treatment.

To study the distribution of expanded activated lymphocytes in severe immunodeficient mice.

Ninety-six NPG mice were used and randomly divided into vehicle control group and administration group. The animals in the vehicle control group were given vehicle by tail vein injection once; the animals in the administration group were given DiR-labeled EAL cells by tail vein injection once. Blood was collected at different time points after administration, and EAL cells were detected by in vivo imaging system. At 1 h, 3 h, 2 d, 7 d, 14 d, 28 d, 42 d, and 56 d after administration, the mice were anesthetized, and blood, heart, liver, spleen, lung, kidney, brain, testis, epididymis, uterus, ovary, stomach, duodenum, colon, bone marrow, fat, and skeletal muscle were taken for cryopreservation. The distribution of EAL cells in peripheral blood and the above-mentioned tissues was studied by flow cytometry, in vivo imaging method and qPCR method.

The numbers of CD₃⁺ T cells, CD₃⁺CD₄⁺ T cells and CD₃⁺CD₈⁺ T cells in peripheral blood were higher at 2 d after administration, and then showed a downward trend, reaching the lowest at 14 d, and maintained a low level thereafter. The results of in vivo imaging showed that the cells were mainly distributed in the lungs, liver, spleen and bilateral leg bones, and were mainly distributed in the lungs and liver within 1 d after administration. The results of gene copy number detection showed that the cells were mainly distributed in the lungs and blood, followed by the spleen, liver and bone marrow, and only a little in other tissues. At 1 h after administration, the gene copy number of cells in the lungs was relatively high. At 2 d, the number of gene copies in the blood and spleen was relatively high, and then gradually decreased, reaching the lowest at 14 d, and at 56 d cells were only detected in individual organs of one mice.

EAL cells were administered to NPG mice, the cells were mainly distributed in lung, blood and spleen. At 56d after administration, the cells were basically eliminated from the mice.

To optimize the preparation and solidification process of paeonol nanosuspensions.

Paeonol nanosuspensions was prepared by solvate-non-solvent precipitation method. With the appearance character, particle size and polymer dispersity index (PDI) as the evaluation indexes, the type of stabilizer, stirring time, stirring speed, drug-auxiliary ratio, organic phase-water ratio and water phase temperature as the influencing factors, the preparation process of nanosuspensions was optimized, and the stability of the prepared nanosuspensions was investigated. The freeze-drying method was used for solidification, and the effect of freeze-drying protectant on solidification was investigated. The drug loading, scanning electron microscopy, differential scanning calorimetry, X-ray diffraction and dissolution in vitro of the solidified nanosuspensions were characterized.

The preparation process of paeonol nanosuspensions was povidone K60 as stabilizer, drug-to-auxiliary ratio 1∶3, organic phase water ratio 1∶10, at 25 ℃, 400 r·min^-1 magnetic stirring for 30 min. The prepared nanosuspensions had a particle size of about 40 nm, with narrow distribution and good stability. The lyophilized protectant was determined to be 5% trehalose. The drug loading capacity of solidified nanosuspensions was 21.63%, and the microscopic surface morphology was spherical, and the drug structure changed from crystalline form to amorphous form. The cumulative dissolution rate of the suspension reached 87.12% in 1 h.

The formulation and preparation process of paeonol nanosuspensions are simple, easy to operate, with good formability after solidification and fast dissolution, which lay a foundation for the development of more dosage forms.