[Determination of concentrations and toxicokinetics of triptolide in plasma and liver of mice by UHPLC-MS/MS].

This study aims to establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) method for determining the concentrations of triptolide(TP) in plasma and liver, and to explore the toxicokinetics of TP and the relationship between TP exposure and liver injury in C57 BL/6 mice, so as to provide reference for dissecting the toxicity mechanism of TP. The liquid chromatography was conducted with ZORBAX SB-C_(18) column(3.0 mm×100 mm, 3.5 µm) and the mobile phase of methanol-0.05 mmol·L~(-1) ammonium acetate. Electrospray ionization(ESI) and multiple reaction monitoring(MRM) mode were employed for mass spectrometry. After oral administration of TP(toxic dose 600 µg·kg~(-1)), the blood and liver tissues of the C57 BL/6 mice were collected at different time points to measure the TP concentrations in plasma and liver tissues. Furthermore, the blood biochemical indexes, including alkaline phosphatase(ALP), alanine aminotransferase(ALT), aspartate aminotransferase(AST), and total bile acid(TBA), were determined. After being processed by DAS 2.0, the experiment data showed that the TP in mice had the toxicokinetic parameters of T_(max)=5 min, C_(max)=14.38 ng·mL~(-1), t_(1/2)=0.76 h, AUC_(0-t)=5.63 h·ng·mL~(-1), MRT_(0-t)=0.56 h, and CL_(Z/F)=103.19 L·h~(-1)·kg~(-1). The trend of TP concentration in mouse liver tissue was consistent with that in plasma. The concentration of TP peaked at the time point of 5 min and then decreased until TP was completely metabolized. The plasma biochemical indexes(ALT, AST, ALP, and TBA) showed no significant changes within 3 h after TP administration. TP had high clearance rate and short residence time and did not significantly increase the blood biochemical indexes in mice. The results suggested that the exposure amount of free TP in vivo cannot directly cause liver injury, which might be caused by the binding of TP to some substances or the stimulation of inflammation and immune response.

RNA interference targeting Ras GTPase gene Ran causes larval and adult lethality in Leptinotarsa decemlineata.

BACKGROUND: RNA interference (RNAi) is a breakthrough technology in pest control. It is highly efficient to Coleopteran pests such as the Colorado potato beetle Leptinotarsa decemlineata, a serious pest defoliator mainly attacking potatoes worldwide. The first step for effective pest control by RNAi is the development of effective and reliable target genes. RESULTS: Our results revealed that continuous ingestion of dsLdRan for 3 days successfully silenced the target gene, inhibited larval growth and killed 100% L. decemlineata larvae. When the bioassay began at the second-, third/fourth-instar larval stages, the larval lethality mainly occurred at the fourth larval instar and prepupal stages, respectively. Importantly, consumption of dsLdRan for 3 days by the newly-emerged males and females effectively knocked down the target transcript, reduced fresh weights and caused 100% of lethality within a week. The LdRan females possessed underdeveloped ovaries. CONCLUSION: Considering that the larvae, adults and eggs are simultaneously sited on the potato plants, bacterially-expressed dsLdRan is a potential RNAi-based strategy for managing L. decemlineata in the potato field. © 2022 Society of Chemical Industry.

[Dynamics and influencing factors of stem diameter micro-variations during the growing season in two typical forestation species in the loess hilly region, China.] / é»„åœŸä¸˜é™µåŒºä¸¤å ¸åž‹é€ æž—æ ‘ç§ç”Ÿé•¿å­£æ ‘å¹²ç›´å¾„å¾®å˜åŒ–åŠ¨æ€åŠå ¶å½±å“å› ç´ .

Using DC3 high-resolution dendrometer and Granier-type thermal dissipation probes, we measured stem diameter micro-variations and xylem sap flow of two typical forestation species, Quercus liaotungensis and Robinia pseudoacacia, for a growing season in the loess hilly region of China. The main environmental factors (soil water content, solar radiation, air temperature and relative humidity) were monitored. The linkage between diameter micro-variations and transpiration water use were analyzed with respect to their responses to environmental factors. The results showed that the variations in stem diameter and sap flux density of both species had obvious diurnal rhythms. The maximum daily shrinkage was positively correlated with daily sap flux density. The micro-variation of stem diameter at the daily scale was affected by transpiration during the day. The maximum daily shrinkage of stem diameter was positively correlated and well fitted with transpiration driving factors (solar radiation, vapor pressure deficit, and the integrated variable VT). The difference in slopes of regression curves suggested that the daily variation of stem diameter in Q. liaotungensis was greater and more sensitive to meteorological factors than that in R. pseudoacacia. The sap flux densities of both tree species were higher during the period with relatively higher soil water content than that with lower soil water content. The difference of maximum daily diameter shrinkage between different soil water conditions was statistically significant in Q. liaotungensis, but not in R. pseudoacacia. These differences may be related to water use strategies, including transpiration regulation and stem water replenishment.

Highly efficient nitrate and phosphorus removal and adsorption of tetracycline by precipitation in a chitosan/polyvinyl alcohol immobilized bioreactor.

Single denitrification using bacteria has been widely investigated, but few studies have focused on the simultaneous removal of nitrate, phosphorus. and tetracycline. Strain L2, an iron-reducing bacteria, was immobilized using chitosan/polyvinyl alcohol to simultaneously remove nitrate and phosphorus. The effects of carbon/nitrogen ratio (1:1, 1.5:1, and 2:1), initial Fe2+ concentration (0, 15, and 30 mg·L-1), and HRT (2, 4, and 6 h) were assessed in bioreactors and optimum conditions were established. Results showed that the nitrate and phosphorus removal efficiency reached 100.00% (2.697 mg·L-1·h-1) and 81.93% (1.533 mg·L-1·h-1) under the conditions of carbon/nitrogen of 2:1, Fe2+ concentration of 30 mg·L-1 and HRT of 6 h. The precipitation of bioreactor, which identified as FeOOH by XRD, had significant adsorption on tetracycline. The results of high-throughput sequencing indicated that strain L2 played a significant role in denitrification. This bioreactor provided effective method for the treatment of polluted water contaminated by nitrate, phosphorus, and tetracycline.

Microbial etiology, susceptibility profile of postradiation nasopharyngeal necrosis patients with nasopharyngeal carcinoma.

OBJECTIVE: Postradiation nasopharyngeal necrosis (PRNN) is a notorious complication after radiotherapy that affects prognosis in patients with nasopharyngeal carcinoma (NPC). It is important for clinical doctors to realize this problem in order to cope with this severe clinical situation. The aim of our study was to assess the bacteriology of PRNN and to demonstrate the antimicrobial susceptibility pattern that should guide the clinicians towards more appropriate antibiotic use. METHODS: Sixty-nine NPC patients with PRNN in our department between March 2013 and December 2017 were retrospectively enrolled. Pathogenic culture and drug sensitivity test were performed in these 69 NPC patients with PRNN. The infection rate of Pathogens and the sensitivity of the drugs were analyzed based on these results. RESULTS: Sixty-nine NPC patients with PRNN were enrolled in our study. Pathogens were identified in 58 (84%) patients. Of the 58 patients, Staphylococcus aureus was isolated in 34 (58.6%) patients. And the second most common group of bacterial isolates was Pseudomonas aeruginosa. Antibiotic sensitivity showed that Levofloxacin was the highest (88.5%), followed by Ciprofloxacin (85.2%) and Gentamicin (80.3%). The only pathologic fungus was Candidaalbicans, about 6.8%. The positive rates of bacterial and fungal culture in PRNN patients were not significantly different from the patients' gender, age, stage, number of radiotherapy courses (P>0.05), but the cure rate was statistically higher in culture-negative patients in comparison with culture-positive patients (63.6% vs 20.7%, P=0.011). CONCLUSION: Our results provide an overall picture of the microbiology and drug susceptibility patterns for NPC patients with PRNN and could help implement guidelines for more rational treatment and improve therapeutic outcome.

1,25-(OH)2D3/Vitamin D receptor alleviates systemic lupus erythematosus by downregulating Skp2 and upregulating p27.

BACKGROUND: Recent evidence has suggested that the 1,25(OH)2D3/Vitamin D receptor (VDR) acts to suppress the immune response associated with systemic lupus erythematosus (SLE), a serious multisystem autoimmune disease. Hence, the aim of the current study was to investigate the mechanism by which 1,25-(OH)2D3/VDR influences SLE through regulating the Skp2/p27 signaling pathway. METHODS: Initially, the levels of 1,25(OH)2D3, VDR, Skp2, and p27 were measured in collected renal tissues and peripheral blood. Meanwhile, the levels of inflammatory factors, biochemical indicators (BUN, Cr, anti-nRNP IgG, anti-dsDNA IgG) and urinary protein levels were assayed in in VDRinsert and VDR-knockout mice in response to 1,25(OH)2D3 supplement. In addition, the distribution of splenic immune cells was observed in these mice. RESULTS: Among the SLE patients, the levels of 1,25(OH)2D3, VDR and p27 were reduced, while the levels of Skp2 were elevated. In addition, the levels of anti-nRNP IgG and anti-dsDNA IgG were increased, suggesting induction of inflammatory responses. Notably, 1,25(OH)2D3/VDR mice had lower concentrations of BUN and Cr, urinary protein levels, precipitation intensity of the immune complex and complement, as well as the levels of anti-nRNP IgG and anti-dsDNA IgG in SLE mice. Additionally, 1,25(OH)2D3 or VDR reduced the degree of the inflammatory response while acting to regulate the distribution of splenic immune cells. CONCLUSION: This study indicated that 1,25-(OH)2D3/VDR facilitated the recovery of SLE by downregulating Skp2 and upregulating p27 expression, suggesting the potential of 1,25-(OH)2D3/VDR as a promising target for SLE treatment.

[Time lag of stem sap flow and its relationships with transpiration characteristics in Quercus liaotungensis and Robina pseudoacacia in the loess hilly region, China]. / é»„åœŸä¸˜é™µåŒºè¾½ä¸œæ Žå’Œåˆºæ§æ ‘å¹²æ¶²æµæ—¶æ»žæ•ˆåº”ä¸Žè’¸è ¾ç‰¹å¾çš„å ³è”æ€§.

Using Granier-type thermal dissipation probes (TDP), we measured stem xylem sap flow of the natural dominant species Quercus liaotungensis and a reforestation species Robinia pseudoacacia from July to September in 2016 in the semiarid loess hilly region. Meteorological factors and soil water content were simultaneously monitored during the study period. Using cross-correlation analysis, time lag between diurnal patterns of sap flux density and vapor pressure deficit (VPD) was quantitatively estimated. Differences in the time lag between the two species and possible influence by different diameter classes and soil water contents were analyzed. The results showed that the diurnal courses of sap flux density were similar to those of meteorological factors, with daily peaks ear-lier than VPD. The peak of VPD lagged behind the sap flux densities of Q. liaotungensis and R. pseudoacacia 118.2 min and 39.5 min, respectively. The peak of PAR lagged behind the sap flux density of Q. liaotungensis 12.4 min, but was 68.5 min ahead of that for R. pseudoacacia. Time lag between sap flux density and VPD significantly varied between tree species and was affected by soil water content. Those during higher soil water content period were about 32.2 min and 68.2 min longer than those during the period with lower soil water content for the two species, respectively. There was no correlation between time lag and tree diameter classes. The time lag between VPD and sap flux density for R. pseudoacacia was about 21.4 min longer in smaller diameter trees than in larger trees, which was significantly different under the lower soil water content. Our results suggested that the time lag effect between VPD and sap flux densities in the two species reflected their sensitivities to driving factors of transpiration, and that higher soil water content was favorable to sap flux density reaching its peak early. The lower soil water content might lead to lower sensitivity of the trees to meteorological factors. R. pseudoacacia was more sensitive to changes of soil water content.

Identification of Cuticular Protein Genes in the Colorado Potato Beetle Leptinotarsa decemlineata (Coleoptera: Chrysomelidae).

Structural cuticular proteins (CPs) are the primary components of insect cuticle, linings of salivary gland, foregut, hindgut and tracheae, and midgut peritrophic membrane. Variation of CPs in insect cuticle can cause penetration resistance to insecticides. Moreover, depletion of specific CP by RNA interference may be a suitable way for the development of potential pest control traits. Leptinotarsa decemlineata (Say) CPs are poorly characterized at present, and therefore, we mined the genome and transcriptome data to better annotate and classify L. decemlineata CPs in this study, by comparison with the annotated CPs of Tribolium castaneum Browse (Coleoptera: Tenebrionidae). We identified 175 CP genes. Except one miscellaneous CP with an 18-amino acid motif, these CPs were classified into 7 families based on motifs and phylogenetic analyses (CPs with a Rebers and Riddiford motif, CPR; CPs analogous to peritrophins, CPAP3 and CPAP1; CPs with a tweedle motif, TWDL; CPs with a 44-amino acid motif, CPF; CPs that are CPF-like, CPFL; and CPs with two to three copies of C-X5-C motif, CPCFC). Leptinotarsa decemlineata CPRs could be categorized into three subfamilies: RR-1 (50), RR-2 (85), and RR-3 (2). The RR-1 proteins had an additional motif with a conserved YTADENGF sequence. The RR-2 members possessed a conserved RDGDVVKG region and three copes of G-x(3)-VV. Few genes were found in TWDL (9), CPAP1 (9), CPAP3 (8), CPF (5), CPFL (4), and CPCFC (2) families. The findings provide valuable information to explore molecular modes of penetration resistance to insecticides and to develop dsRNA-based control method in L. decemlineata.

Ginnalin A from Kujin tea (Acer tataricum subsp. ginnala) exhibits a colorectal cancer chemoprevention effect via activation of the Nrf2/HO-1 signaling pathway.

Ginnalin A (also known as acertannin) is one of the most important phenolic compounds of several beverage Acer plants. In this study, it is reported for the first time that ginnalin A is an activator of the Nrf2 signaling pathway in human colon cancer cells. Ginnalin A, isolated from the leaves of Acer tataricum subsp. ginnala, exhibited promising preventive activity against colon cancer cells (HCT116, SW480 and SW620) with IC50 values of 24.8 µM, 22.0 µM and 39.7 µM, respectively. In addition, it significantly reduced the colony formation of these cells. Flow cytometry analysis indicated that ginnalin A suppressed cancer proliferation via the induction of cell cycle arrest at the S-phase. Real time PCR analysis demonstrated that ginnalin A can upregulate the mRNA expression levels of Nrf2-related antioxidant genes Nrf2, HO-1 and NQO1. Western blotting analysis revealed that ginnalin A promoted the Nrf2 nuclear translocation and upregulated the proteins Nrf2, HO-1 and NQO1. Moreover, the upregulation of p62 and the inhibition of Keap1 were also found by Western blotting analysis. Therefore, the activation of the Nrf2 signaling pathway was probably induced through the upregulation of p62 and the inhibition of Keap1.

Knocking down a putative Δ(1) -pyrroline-5-carboxylate dehydrogenase gene by RNA interference inhibits flight and causes adult lethality in the Colorado potato beetle Leptinotarsa decemlineata (Say).

BACKGROUND: Leptinotarsa decemlineata is an able disperser by flight. Novel control strategies must be explored to control the damage and inhibit the dispersal efficiently. Proline is a major energy substrate during flight. Δ-Pyrroline-5-carboxylate dehydrogenase (P5CDh) catalyses the second step of proline degradation for the production of ATP. RESULTS: A full-length Ldp5cdh cDNA was cloned. Ldp5cdh was ubiquitously expressed in the eggs, the first through fourth larval instars, wandering larvae, pupae and adults. In the adults, Ldp5cdh mRNA was widely distributed in thorax muscles, midgut, foregut, hindgut, Malpighian tubules, ventral ganglion, fat body and epidermis, with the expression levels from the highest to the lowest. Two double-stranded RNAs (dsRNAs) (dsLdp5cdh1 and dsLdp5cdh2) targeting Ldp5cdh were constructed and bacterially expressed. Ingestion of dsLdp5cdh1 and dsLdp5cdh2 successfully silenced Ldp5cdh, significantly increased the contents of proline, arginine and alanine, but strongly decreased the contents of asparate, asparagine, glutamate and glutamine in the haemolymph. Moreover, knocking down Ldp5cdh significantly reduced ATP content, decreased flight speed, shortened flight distance and increased adult mortality. CONCLUSIONS: It seems that identified Ldp5cdh encodes a functional P5CDh enzyme, and Ldp5cdh may serve as a potential target for dsRNA-based pesticide for controlling the damage and dispersal of L. decemlineata adults. © 2014 Society of Chemical Industry.

A putative Δ1-pyrroline-5-carboxylate synthetase involved in the biosynthesis of proline and arginine in Leptinotarsa decemlineata.

Delta 1-pyrroline-5-carboxylate synthetase (P5CS) catalyzes the conversion of glutamate (Glu) to Glu semialdehyde (GSA). GSA spontaneously cyclizes to form P5C. P5C is then reduced to proline (Pro) or is converted to ornithine, the intermediate for arginine (Arg) biosynthesis. In the present study, a full-length Ldp5cs complementary DNA was cloned from the Colorado potato beetle Leptinotarsa decemlineata, a notorious insect defoliator of potato in most potato-growing regions of the world. Ldp5cs encodes a 792-amino-acid protein which shares high identity to homologues from other insect species. Quantitative reverse transcription polymerase chain reaction revealed that Ldp5cs was ubiquitously expressed in the eggs, first to fourth-instar larvae, wandering larvae, pupae and sexually mature adults. In the adults, Ldp5cs mRNA levels were higher in the fat body, foregut, midgut and hindgut, moderate in the ventral ganglion, lower in the thorax muscles, epidermis and Malpighian tubules. Two double-stranded RNAs (dsRNAs) (dsLdp5cs1 and dsLdp5cs2) targeting Ldp5cs were constructed and bacterially expressed. Ingestion during 3 consecutive days of dsLdp5cs1 or dsLdp5cs2 successfully silenced Ldp5cs, significantly reduced the contents of Pro and Arg in the hemolymph, decreased flight speed and shortened flight distance of the resulting adults. Furthermore, knocking down Ldp5cs significantly increased adult mortality. Thus, our results suggest that identified Ldp5cs encodes a functional P5CS enzyme that is involved in the biosynthesis of Pro and Arg in L. decemlineata.

Extracts of Celastrus orbiculatus exhibit anti-proliferative and anti-invasive effects on human gastric adenocarcinoma cells.

OBJECTIVE: To assess the effect of Celastrus orbiculatus (COE) on growth, invasion and migration of human gastric cancer MGC-803 cells and to explore the possible mechanism. METHODS: The effect of COE on cell viability, apoptosis, adhesion, invasion and migration were studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, flow cytometric, cell adhesion and transwell assay, respectively. The activity and expression of matrix metalloproteinase-9 (MMP-9) were determined by gelatin zymography, Western blot and quantitative real-time polymerase chain reaction analysis. Meanwhile, effects of COE on the expression of mitogen-activated protein kinases (MAPKs), serine threonine kinase (Akt), nuclear factor κB (NF-κB) were investigated with Western blot analysis. RESULTS: COE inhibited proliferation and induced apoptosis of MGC-803 cells in a dose-dependent manner. When treated with low-toxic (below 80 µg/mL) doses of COE, cell adhesion, invasion and migration were markedly suppressed. Furthermore, the gelatinolytic activity and expression of MMP-9 were also remarkably suppressed in a dose-dependent manner. In addition, upstream signaling pathways, including the phosphatidylinositol-3 kinase (PI3K)/Akt and NF-κB, were suppressed by COE. Additionally, the PI3K/Akt inhibitor, LY294002, in treating MGC-803 cells potently suppressed cell invasion and migration as well as expression of MMP-9. Similarly, the combined treatment with COE and LY294002 showed a synergistic effect compared with the treatment with COE or LY294002 alone in MGC-803 cells. CONCLUSIONS: COE inhibits invasion and migration of MGC-803 cells by reducing MMP-9 expression. It also inhibit PI3K/Akt and NF-κB signaling pathways, which may offer a novel approach for the treatment of human gastric cancer.

Human serum albumin-based design of a diflunisal prodrug.

The cyclooxygenase-2 inhibitor, diflunisal, is used in the clinic for its anti-inflammatory activity. About 99% of a dose of diflunisal is unavailable for reaction with the target enzyme, because diflunisal strongly binds to human serum albumin (HSA). To reduce the binding affinity of diflunisal to albumin, we designed and synthesized the prodrug acetyldiflunisal. The crystal structure of HSA complexed with fatty acid and acetyldiflunisal revealed that acetyldiflunisal binds to the IIA subdomain and that upon binding, it acetylates lysine 199. Mass spectrometry confirmed that acetyldiflunisal acetylates Lys199. The acetylated albumin had twofold weaker binding affinity for diflunisal as demonstrated by fluorescence quenching. Reduced binding affinity means that diflunisal is more easily released from acetylated albumin into the circulation. Therefore, lower doses of acetyldiflunisal compared to diflunisal will be required. Taken together, our results not only provide a template for design of HSA-based prodrugs, but also pave the way toward more effective use of diflunisal in the clinic.

Celastrus orbiculatus extract inhibits tumor angiogenesis by targeting vascular endothelial growth factor signaling pathway and shows potent antitumor activity in hepatocarcinomas in Vitro and in Vivo.

OBJECTIVE: Celastrus orbiculatus Thunb. has been used for thousands of years in China as a remedy against cancer and inflammatory diseases. This study aims to investigate whether C. orbiculatus extract (COE) could inhibit angiogenesis, which is the pivotal step in tumor growth, invasiveness, and metastasis. METHODS: In this study, the extract from the stem of C. orbiculatus was used. Mouse hepatic carcinoma cells (Hepa1-6) were treated with COE in different nontoxic concentrations (10, 20, 40, 80, and 160 µg/mL). The mRNA and protein expression levels of vascular endothelial growth factor (VEGF) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively; the active fractions were further tested on C57BL/6 mice and human umbilical vein endothelial cells (HUVEC) for any antiangiogenic effects. RESULTS: COE significantly inhibited proliferation and induced apoptosis in Hepa1-6 cells and inhibited VEGF expression at both mRNA and protein levels. Furthermore, this agent inhibited the formation of the capillary-like structure in primary cultured HUVEC in a dose-dependent manner. In vivo, COE significantly reduced the volume and weight of solid tumors with low adverse effects and decreased tumor angiogenesis. CONCLUSIONS: In summary, COE could be used to treat hepatic carcinoma. The mechanisms of the antitumor activity of COE may be due to its effects against tumor angiogenesis by targeting the VEGF protein.

Rosiglitazone reverses mitomycin C resistance in human gastric cancer cells.

INTRODUCTION: To explore the mechanisms of rosiglitazone (ROS), a selective peroxisome proliferator-activated receptor gamma ligand, in reversing mitomycin C (MMC) resistance in a human drug-resistant gastric cancer cell line. METHODS: The vincristine-resistant human gastric cancer cell line SGC7901/VCR and its parental cell line SGC7901 were treated with ROS, MMC (negative control), cyclosporine A+MMC (positive control) or ROS+MMC. A tetrazolium blue (methyl thiazolyl tetrazolium) assay was used to evaluate the sensitivity to these treatments. Flow cytometry analysis and acridine orange-ethidium bromide (AO-EB) fluorescent staining were used to determine the effects of ROS on MMC-induced apoptosis. Reverse transcription polymerase chain reaction and western blotting were used to measure the expression of multidrug resistant 1 (MDR1), Livin and P-glycoprotein (P-gp). RESULTS: ROS administration dose dependently increased the reversal index in MMC-treated SCG7901/VCR cells. ROS increased apoptosis in SGC7901/VCR cells compared with the blank group and MMC group. ROS+MMC also increased apoptosis in SGC7901/VCR cells compared with other groups (P < 0.05 or P < 0.01). The mRNA expression of MDR1 and Livin and the protein expression of P-gp in SGC7901/VCR cells were significantly higher than those in SGC7901 cells (P < 0.01). However, ROS or ROS+MMC treatment markedly upregulated the mRNA expression of MDR1 and Livin and the protein expression of P-gp in SGC7901/VCR cells (P < 0.01). CONCLUSIONS: ROS reverses MMC resistance in human gastric cancer SGC7901/VCR cells by reducing expression of MDR1, Livin and P-gp and increasing apoptosis.

[Effects of some traditional Chinese drugs on Mdr1 gene and its expression product in K562/A02 cells].

This study was purposed to investigate the effects of angelicasinensis (Oliv) Diel compound injection, vauqueline, ephedrine and strychnine on human erythroleukemia multidrug resistance (MDR) K562/A02 cell line Mdr1 gene and p-glycoprotein. The MTT and trypan blue methods were used to analyze the cytotoxic effect of above-mentioned traditional Chinese drug; the expressions of K562/A02 cells Mdr1 gene and p-glycoprotein were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot respectively. The results showed that after K562/A02 cell were treated with angelicasinensis (Oliv) diels compound injection, vauqueline and ephedrine, mRNA transcription of Mdr1 gene was reduced significantly (p < 0.01); the expression of P-gp also decreased (p < 0.01). The expression level P-gp in group treated with vauqueline was the lowest, but the Mdr1 mRNA level and expression of P-gp of K562/A02 cells treated with strychnine did not obviously changed. It is concluded that angelicasinensis (Oliv) diels compound injection, vauqueline, ephedrine can partly reverse the multidrug resistance of K562/A02 cells, the down-regulation of Mdr1 gene causing decrease of p-glycoprotein expression may be one of the MDR reversal mechanisms in K562/A02 cells.

[Clinical observation on treatment of male infertility with positive antisperm antibody by self-formulated Xiaokang Zhongzi Recipe].

OBJECTIVE: To observe the therapeutic effect of self-formulated Xiaokang Zhongzi Recipe (XZR) in treating male infertility with positive antisperm antibody (AsAb). METHODS: One hundred and ten male infertility patients with positive AsAb were randomly assigned, according to randomized digital table, to the trial group treated with XZR, and the control group, treated with prednisone, 55 in each group. Clinical therapeutic effect and adverse reactions of the drugs were observed after 3 months of treatment. RESULTS: In the trial group after treatment, 38 patients (69.1%) were cured, 14 (25.4%) improved and 3 treated in vain (5.5%, including 2 dropped out). While in the control group, the corresponding numbers were 12 (21.8%), 11 (20.0%) and 32 (58.2%, including 4 dropped out, 20 remained AsAb positive and 8 with AsAb reverted to positive after negative conversion). The negative conversion rate and average negative conversion time in the trial group and the control group were respectively 94.5%, (45 +/- 14) days and 41.8%, (62 +/- 21) days. Significant difference between the two groups was shown in therapeutic effectiveness and average negative conversion time (P < 0.01). Adverse reactions occurred in 3 cases (5.5%) in the trial group and 8 (14.5%) in the control group. CONCLUSION: XZR is better than prednisone in treating male infertility with positive AsAb, and with fewer and milder adverse reactions.