Benzylisoquinoline alkaloids (BIAs) are a structurally diverse group of plant metabolites renowned for their pharmacological properties. However, sustainable sources for these compounds remain limited. Consequently, researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches. CYP80B, a family of cytochrome P450 (CYP450) enzymes, plays a crucial role in BIA biosynthesis. Previously reported CYP80Bs are known to catalyze the 3'-hydroxylation of (S)-N-methylcoclaurine, with the N-methyl group essential for catalytic activity. In this study, we successfully cloned a full-length CYP80B gene (StCYP80B) from Stephania tetrandra (S. tetrandra) and identified its function using a yeast heterologous expression system. Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that StCYP80B could catalyze N-methylcoclaurine and coclaurine into their respective 3'-hydroxylated products. Notably, StCYP80B exhibited an expanded substrate selectivity compared to previously reported wild-type CYP80Bs, as it did not require an N-methyl group for hydroxylase activity. Furthermore, StCYP80B displayed a clear preference for the (S)-configuration. Co-expression of StCYP80B with the CYP450 reductases (CPRs, StCPR1, and StCPR2), also cloned from S. tetrandra, significantly enhanced the catalytic activity towards (S)-coclaurine. Site-directed mutagenesis of StCYP80B revealed that the residue H205 is crucial for coclaurine catalysis. Additionally, StCYP80B exhibited tissue-specific expression in plants. This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.
Cytochrome P-450 Enzyme System/chemistry* ; Benzylisoquinolines/chemistry* ; Hydroxylation ; Plant Proteins/chemistry* ; Alkaloids/metabolism* ; Stephania tetrandra/genetics*

Objective:To compare the clinical outcomes of capsular closure versus those of non-closure in hip arthroscopy for femoroacetabular impingement (FAI) through a meta-analysis of randomized controlled trials (RCTs).Methods:A systematic search was conducted in Cochrane Library, PubMed, Web of Science, China National Knowledge Infrastructure (CNKI), and Wanfang Database for RCTs comparing capsular closure with non-closure in hip arthroscopy for FAI, covering the period from database inception to October 2024. A meta-analysis was performed using RevMan 5.3 software to compare outcomes between patients undergoing hip arthroscopy with capsular closure versus those without capsular closure within 2 years postoperatively. The following parameters were evaluated: the modified Harris hip score (mHHS), 12-item International Hip Outcome Tool (iHOT-12), hip outcome score-activities of daily living (HOS-ADL), hip outcome score-sport specific (HOS-SSS), Copenhagen hip and groin outcome score (HAGOS), visual analog scale (VAS) for pain, reoperation rate, complication rate, and rate of patient satisfaction.Results:A total of 5 RCTs involving 432 patients were included, with 215 cases in the capsular closure group and 217 cases in the non-closure group. The follow-up duration for the patients in the included studies ranged from 12 to 24 months. Meta-analysis revealed no significant differences between the capsular closure and non-closure groups in postoperative functional scores (mHHS, iHOT-12, HOS-SSS, HAGOS), VAS pain score, reoperation rate, complication rate, or rate of patient satisfaction ( P>0.05). The capsular closure group demonstrated significantly better HOS-ADL at 2 years postoperatively than the non-closure group (MD=-3.57, 95% CI: -5.86 to -1.28, P=0.002). Conclusion:In patients with FAI undergoing hip arthroscopy, compared to the non-closure, capsular closure leads to significant improvements in mid-term daily activities, but similar outcomes in short-term hip function, pain control, reoperation rate, and complication incidence.

Objective:To compare the clinical outcomes of capsular closure versus those of non-closure in hip arthroscopy for femoroacetabular impingement (FAI) through a meta-analysis of randomized controlled trials (RCTs).Methods:A systematic search was conducted in Cochrane Library, PubMed, Web of Science, China National Knowledge Infrastructure (CNKI), and Wanfang Database for RCTs comparing capsular closure with non-closure in hip arthroscopy for FAI, covering the period from database inception to October 2024. A meta-analysis was performed using RevMan 5.3 software to compare outcomes between patients undergoing hip arthroscopy with capsular closure versus those without capsular closure within 2 years postoperatively. The following parameters were evaluated: the modified Harris hip score (mHHS), 12-item International Hip Outcome Tool (iHOT-12), hip outcome score-activities of daily living (HOS-ADL), hip outcome score-sport specific (HOS-SSS), Copenhagen hip and groin outcome score (HAGOS), visual analog scale (VAS) for pain, reoperation rate, complication rate, and rate of patient satisfaction.Results:A total of 5 RCTs involving 432 patients were included, with 215 cases in the capsular closure group and 217 cases in the non-closure group. The follow-up duration for the patients in the included studies ranged from 12 to 24 months. Meta-analysis revealed no significant differences between the capsular closure and non-closure groups in postoperative functional scores (mHHS, iHOT-12, HOS-SSS, HAGOS), VAS pain score, reoperation rate, complication rate, or rate of patient satisfaction ( P>0.05). The capsular closure group demonstrated significantly better HOS-ADL at 2 years postoperatively than the non-closure group (MD=-3.57, 95% CI: -5.86 to -1.28, P=0.002). Conclusion:In patients with FAI undergoing hip arthroscopy, compared to the non-closure, capsular closure leads to significant improvements in mid-term daily activities, but similar outcomes in short-term hip function, pain control, reoperation rate, and complication incidence.

Nanocrystal technology changes the solubility and dissolution rate of insoluble drugs by reducing the particle size to the nanometer level.This technology is not limited by carrier materials and encapsulation rate.It is suitable for a variety of drug delivery routes and easy for industrial production.It has gradually become a cutting-edge hot technology in the international pharmaceutical field to improve the absorption of insoluble drugs and improve their bioavailability.This article introduces the influencing factors and challenges of nanocrystal drugs(NC)in parenteral,ocular,transdermal and pulmonary administration,and focuses on nanocrystal drugs that have been marketed or are still in the preclinical or clinical trial stages of these delivery systems,in order to provide some insight for the further development of poorly soluble drug nanocrystal preparations.

Objective:To compare the short-term efficacy of hip arthroscopic surgery assisted by platelet-rich plasma (PRP) and hip arthroscopy alone in the treatment of femoroacetabular impingement (FAI).Methods:A retrospective cohort study was performed on the clinical data of 133 FAI patients admitted to Fourth Medical Center of PLA General Hospital from January 2019 to January 2021. The patients included 86 males and 47 females, aged 19-71 years [(39.1±12.6)years]. A total of 67 patients were treated with hip arthroscopy alone (hip arthroscopy group), and 66 patients were treated with PRP after hip arthroscopy under ultrasound guidance (hip arthroscopy+PRP group). The two groups were compared before, at 12 months after surgery and at the last follow-up regarding the following items: Visual Analogue Scale (VAS), Modified Harris Hip Score, International Hip Outcome Tool-12 (iHOT-12), and Hip Outcome Score Activities of Daily Living Scale (HOS-ADL). The incidence rate of complications after surgery was compared between the two groups.Results:A total of 108 patients were followed up for 24-36 months [(28.5±3.8)months], while 25 patients were lost to follow-up because of withdrawal of consent, wrong telephone number, etc, including 11 patients (16.4%) in the hip arthroscopy group and 14 patients (21.2%) in the hip arthroscopy+PRP group. The values of VAS in the hip arthroscopy group before, at 12 months after surgery and at the last follow-up were 5.00(5.00, 7.00)points, 3.00(2.00, 3.75)points, and 1.00(0.00, 2.00)points, respectively; the values of Modified Harris Hip Score were 49.00(39.00, 57.00)points, 76.00(69.25, 82.00)points, and 86.00(82.00, 88.00)points, respectively; the values of iHOT-12 were 0.45(0.28, 0.58)points, 0.69(0.58, 0.80)points, and 0.81(0.70, 0.92)points, respectively; the values of HOS-ADL were 0.52(0.42, 0.68)points, 0.87(0.75, 0.93)points, and 0.93(0.86, 0.99)points, respectively. The scores of VAS in the hip arthroscopy + PRP group before, at 12 months after surgery and at the last follow-up were 6.00(5.00, 7.00)points, 3.00(2.00, 3.75)points, and 1.00(0.00, 2.00)points, respectively; the values of Modified Harris Hip Score were 46.50(37.00, 56.75)points, 78.00(72.00, 84.00)points, and 84.50(82.00, 88.00)points, respectively; the values of iHOT-12 were 0.42(0.26, 0.51)points, 0.66(0.58, 0.74)points, and 0.81(0.68, 0.88)points, respectively; the values of HOS-ADL were 0.54(0.38, 0.65)points, 0.87(0.72, 0.96)points, and 0.94(0.86, 1.00)points, respectively. In both groups, VAS, Modified Harris Hip Score, iHOT-12, and HOS-ADL were significantly improved at 12 months after surgery and at the last follow-up compared with those before surgery, and were further improved at the last follow-up compared with those at 12 months after surgery (all P<0.01). There were no significant differences in VAS, Modified Harris Hip Score, iHOT-12 and HOS-ADL between the two groups before, at 12 months after surgery and at the last follow-up (all P>0.05). There was no significant difference in the incidence rates of postoperative hip pain and clicking between the two groups (both P>0.05). Conclusion:Hip arthroscopy can considerably improve short-term hip symptoms and function in FAI patients, but the use of PRP treatment after hip arthroscopy cannot further improve its short-term efficacy in FAI patients.

Alcohol abuse leads to alcoholic liver disease and no effective therapy is currently available. Wuzhi Tablet (WZ), a preparation of extract fromthat is a traditional hepato-protective herb, exerted a significant protective effect against acetaminophen-induced liver injury in our recent studies, but whether WZ can alleviate alcohol-induced toxicity remains unclear. This study aimed to investigate the contribution of WZ to alcohol-induced liver injury by using chronic-binge and acute models of alcohol feeding. The activities of ALT and AST in serum were assessed as well as the level of GSH and the activity of SOD in the liver. The expression of CYP2E1 and proteins in the NRF2-ARE signaling pathway including NRF2, GCLC, GCLM, HO-1 were measured, and the effect of WZ on NRF2 transcriptional activity was determined. We found that both models resulted in liver steatosis accompanied by increased transaminase activities, but that liver injury was significantly attenuated by WZ. WZ administration also inhibited CYP2E1 expression induced by alcohol, and elevated the level of GSH and the activity of SOD in the liver. Moreover, the NRF2-ARE signaling pathway was activated by WZ and the target genes were all upregulated. Furthermore, WZ significantly activated NRF2 transcriptional activity. Collectively, our study demonstrates that WZ protected against alcohol-induced liver injury by reducing oxidative stress and improving antioxidant defense, possibly by activating the NRF2-ARE pathway.

To summarize the clinical experience of Professor WANG Meng-yong in the treatment of uric acid nephropathy. Professor WANG believes that the disease is mainly caused by spleen and kidney deficiency, disorder of function of Sanjiao, and pathological products, such as phlegm dampness and blood stasis and other metabolic disorders. Therefore, the treatment should distinguish symptoms and essence. Starting from pathogenesis and pathological features of spleen and kidney deficiency and phlegm dampness and blood stasis, the treatment should flexibly apply the methods of nourishing spleen and kidney, reducing phlegm and dispelling humidity, and activating blood and using diuretic of hydragogue to alleviate water retention, which can greatly reduce side effects caused by the long-term use of Western medicine and the onset of gout, and then to help disease recovery.

At present,the study of intestinal absorption of oral drugs mainly includes in vitro,in vivo and in situ methods.In view of the advantages of in situ intestinal perfusion such as simple operation,mature technology,controllable,ensure the neuroendocrine regulation and blood supply,and so on,which could better reflect the true situation of drug absorption.In this study,the research methods and characteristics of intestinal absorption of oral drugs were systematically introduced.The recirculating perfusion method and single-pass perfusion method were compared,and several volume correction methods were also introduced.In order to ensure the operability and accuracy of experimental results,proper experiment method of intestinal absorption will be adopt according to the factors such as drug characters,experiment requirements,experimental conditions,and so on.The article provides a scientific basis for the development of pharmaceutical dosage and clinical rational drug use.

Objective Objective To evaluate the efficacy of Heqi San combined with metformin in the treatment of obese polycystic ovary syndrome (PCOS). Methods A total of 60 patients with obese polycystic ovary syndrome who met the inclusion criteria were randomly divided into 2 groups, 30 patients in each group. The control group was treated with metformin, and the treatment group was treated with Heqi San on the basis of the control group. Both groups were treated for 3 months. The FPG was determined by glucose oxidase method, and fasting insulin (FINS), TC, TG, LDL-C, HDL-C level were measured by radio immunoassay, and the insulin resistance index (IR) was measured by the method of HOMA-IR. The luteotropic hormone (LH), pro follicle stimulating hormone (FSH) and testosterone (T) were detected by the automatic electrochemical light detector, and the ratio of LH/FSH was calculated. The ovarian volume and body mass index changes and clinical effect were evaluated. Results After treatment, the BMI (20.09 ± 3.12 kg/cm2 vs. 23.39 ± 1.43 kg/cm2, t=6.889), FPG (4.44 ± 0.17 mmol/L vs. 4.74 ± 0.16 mmol/L, t=6.945), FINS (10.69 ± 2.41 IU/L vs. 16.29 ± 5.95 IU/L, t=4.778), TC (3.91 ± 0.50 mmol/L vs. 4.20 ± 0.39 mmol/L, t=2.505), LDL-C (2.64 ± 0.63 mmol/L vs. 3.18 ± 0.62 mmol/L, t=3.346), LH (8.70 ± 2.44 U/L vs. 10.27 ± 2.69 U/L, t=2.934), the ratio of LH/FSH (1.33 ± 0.58 vs. 2.18 ± 0.56, t=5.775), T (1.73 ± 0.74 nmol/L vs. 2.95 ± 1.10 nmol/L, t=5.040) and ovarian volume (12.61 ± 2.29 mm3 vs. 14.26 ± 2.52 mm3, t=6.982) in the treatment group were significantly lower than those in the control group (P<0.01 or P<0.05). The FSH (7.33 ± 2.28 U/L vs. 5.95 ± 1.20 U/L, t=2.934) in the treatment group were significantly higher than those in the control group (P<0.05). The total effective rate in the treatment group was 83.3% (25/30) and 63.3% (19/30) in the control group. The difference between the 2 groups was statistically significant (χ2=3.068, P=0.050). Conclusions The Heqi San combined with metformin can effectively improve the body mass index, glucose metabolism, lipid metabolism index and sex hormone level, and reduce the volume of ovary in patients with PCOS, and the curative effect is better than that of oral metformin alone.

Mammalian pancreatic β-cells play a pivotal role in development and glucose homeostasis through the production and secretion of insulin. Functional failure or decrease in β-cell number leads to type 2 diabetes (T2D). Despite the physiological importance of β-cells, the viability of β-cells is often challenged mainly due to its poor ability to adapt to their changing microenvironment. One of the factors that negatively affect β-cell viability is high concentration of free fatty acids (FFAs) such as palmitate. In this work, we demonstrated that Yes-associated protein (Yap1) is activated when β-cells are treated with palmitate. Our loss- and gain-of-function analyses using rodent insulinoma cell lines revealed that Yap1 suppresses palmitate-induced apoptosis in β-cells without regulating their proliferation. We also found that upon palmitate treatment, re-arrangement of F-actin mediates Yap1 activation. Palmitate treatment increases expression of one of the Yap1 target genes, connective tissue growth factor (CTGF). Our gain-of-function analysis with CTGF suggests CTGF may be the downstream factor of Yap1 in the protective mechanism against FFA-induced apoptosis.
Actins ; metabolism ; Adaptor Proteins, Signal Transducing ; antagonists & inhibitors ; genetics ; metabolism ; Animals ; Apoptosis ; drug effects ; physiology ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Line, Tumor ; Connective Tissue Growth Factor ; genetics ; metabolism ; pharmacology ; Cytochalasin D ; pharmacology ; Fatty Acids, Nonesterified ; pharmacology ; HEK293 Cells ; Humans ; Immunohistochemistry ; Insulin-Secreting Cells ; cytology ; drug effects ; metabolism ; Mice ; Microscopy, Fluorescence ; Palmitic Acid ; pharmacology ; Phosphoproteins ; antagonists & inhibitors ; genetics ; metabolism ; RNA Interference ; RNA, Small Interfering ; metabolism ; Rats ; Recombinant Proteins ; genetics ; metabolism ; pharmacology ; Thiazolidines ; pharmacology