Non-clinical reproductive toxicity studies typically employ mammals like rats, rabbits, and cynomolgus monkeys, with animal pregnancy being a key challenge in such testing. This article focuses on the difficulties encountered in the animal pregnancy process and potential countermeasures. Rats can be used for fertility and early embryonic development toxicity studies (Segment Ⅰ), embryo-fetal development toxicity studies (Segment Ⅱ), and perinatal toxicity studies (Segment Ⅲ). The estrous cycle of female rats can be determined by vaginal smear, and mating behavior is confirmed through copulatory plug checks the following day after pairing one female with one male in the same cage. Rabbits are commonly used in embryo-fetal development toxicity studies (Segment Ⅱ). Mating behavior between male rabbits and estrous females is observed to determine the time of conception. However, challenges such as atypical estrus of female rabbits, large variations in estrus between batches, and mating failure often occur in reproductive toxicity testing, which may be addressed through prolonged light exposure, increased protein supplementation, optimized mating strategies, and environmental modifications like female and male rabbits are raised adjacent to each other. Non-human primates (NHPs) are typically employed in perinatal toxicity studies (Segment Ⅲ), where one of the key challenges lies in accurately determining sexual maturity in males - a critical factor for reproductive toxicity testing, which can be assessed through comprehensive evaluation of age, body weight, and testicular volume. Generally, male macaques are considered sexually mature when they meet the following criteria: age >4.5 years, body weight >4.5 kg, single testis volume >10 mL, and combined testicular volume >20 mL. For pregnancy confirmation, ultrasound examination demonstrating visible gestational sacs is required, though this necessitates experienced veterinary clinicians to establish standardized ultrasound examination protocols. In conclusion, reproductive toxicity studies should employ species-appropriate detection methods and evaluation criteria based on anatomical characteristics of the reproductive system to ensure successful mating and proper study execution.

ObjectiveTo characterize the changes in the overall chemical profile and key index components during nine-time repeating steaming and sun-drying processing of Rhei Radix et Rhizoma， and to reveal the change law of its material basis. MethodsHigh performance liquid chromatography（HPLC） was used to analyze the changes in the overall chemical profile of Rhei Radix et Rhizoma decoction pieces， and the contents of 15 main active components such as chrysophanol-8-O-β-D-glucoside， chrysophanol and gallic acid in the process of nine-time repeating steaming and sun-drying were determined. Combined with chemometrics， the contents and quantity ratio relationships of the glycosides， aglycones and tannins during the processing of Rhei Radix et Rhizoma were analyzed， and the partial least squares-discriminant analysis（PLS-DA） and cluster analysis of the main components in different steaming times were conducted， the statistically significant differential markers were selected with the variable importance in the projection（VIP） value>1. ResultsIn the nine-time repeating steaming and sun-drying process of Rhei Radix et Rhizoma， there were certain regularity in the number and peak area of characteristic peaks and the steaming and sun-drying times， the anthraquinone glycosides and aglycones could be roughly divided into three stages， including rapid change stage， fluctuation change stage and stable stage， and the total amount of tannins showed a decreasing trend. However， the ratios between the three components mentioned above tended to stabilize after five rounds of steaming and sun-drying. The results of PLS-DA and cluster heatmap showed that the content of each component in Rhei Radix et Rhizoma fluctuated greatly during the 1-4 steaming and sun-drying processes， while the content of each component was relatively close during the 5-9 steaming and sun-drying processes. After screening， it was found that chrysophanol， emodin， chrysophanol-8-O-β-D-glucoside， rhein， physcion and emodin-8-O-β-D-glucoside could be used as the index components for distinguishing the processed products of Rhei Radix et Rhizoma with different steaming and sun-drying times. ConclusionThe changes in the properties and efficacy of Rhei Radix et Rhizoma caused by the processing of nine-time repeating steaming and sun-drying are due to the changes in the composition and ratio of various glycosides and complex tannins in this herb， which is also the key to the formation of its characteristic of "purgation with supplement". This study can provide a basis for the research on the processing mechanism of Rhei Radix et Rhizoma and the establishment of processing specifications.

ObjectiveTo investigate the efficacy and mechanism of saffron floral bio-residues（SFB） in the treatment of hyperuricemia（HUA） combined with gouty arthritis（GA） in a compound compatibility setting. MethodScreening candidate control Chinese medicines for compound and SFB based on network target distance calculation and data analysis. After adaptive feeding of 80 SD rats for 7 days， 10 rats were randomly selected as the blank group， while the remaining 70 rats were intraperitoneally injected with 3% potassium oxonate and orally administered with 1% adenine for 14 consecutive days. On the 13^th day， rats were injected with 2.5% sodium urate solution into the right ankle joint cavity to induce swelling of the joint capsule on the opposite side， inducing a HUA combined with GA model. At the same time， the modeling rats were randomly divided into 7 groups， including the model group， benzbromarone group（positive drug， 0.02 g·kg^-1）， Tongfengshu tablets group（9 g·kg^-1）， Tongfengshu granules group（9 g·kg^-1）， SFB granules group（3.6 g·kg^-1）， Plantaginis Semen granules group（3.6 g·kg^-1）， and new formula group（SFB replacing Plantaginis Semen in Tongfengshu granules， 9 g·kg^-1）， with 10 rats in each group. Each treatment group was orally administered with the corresponding drugs according to body weight， while the control and model groups were given equal volume of distilled water by gavage once a day for 14 consecutive days. After 14 days of synchronous administration and modeling， changes in gait， ankle joint swelling and mechanical pain threshold in rats were observed， and serum uric acid， creatinine， urea nitrogen and xanthine oxidase（XOD） were measured. Enzyme-linked immunosorbent assay（ELSIA） was used to detect the levels of tumor necrosis factor（TNF）-α， interleukin（IL）-1β and IL-6 in rat serum， hematoxylin-eosin（HE） staining was used to observe the pathological changes in the liver， kidney and ankle joints of rats， Western blot was used to detect the expression levels of uric acid transporter 1（URAT1）， glucose transporter 9 （GLUT9）， organic anion transporter 1（OAT1）， adenosine triphosphate（ATP） binding cassette transporter G2（ABCG2）， and liver XOD proteins. ResultThrough network pharmacology analysis， Plantaginis Semen was selected as a candidate control herb， and Tongfengshu tablets was used as a compound compatibility environment to explore the efficacy of SFB in reducing blood uric acid levels and treating GA. Animal experiments showed that compared with the blank group， the gait score and joint swelling degree of the model group were significantly increased， and the mechanical pain threshold was significantly decreased（P<0.01）. Compared with the model group， the gait score， joint swelling degree and mechanical pain threshold of rats in each medication group were improved to varying degrees. Biochemical indicators showed that compared with the blank group， the serum uric acid， creatinine， urea nitrogen and XOD levels of the model group were significantly increased（P<0.01）. Compared with the model group， the serum uric acid and XOD levels of rats in each treatment group were significantly decreased（P<0.01）. ELISA results showed that compared with the blank group， the levels of serum TNF-α， IL-1β and IL-6 in the model group were significantly increased（P<0.01）. Compared with the model group， the levels of TNF-α， IL-1β and IL-6 in the benzbromarone group， Tongfengshu tablets group， Tongfengshu granules group and new formula group were significantly reduced（P<0.05，P<0.01）. Western blot results showed that compared with the blank group， the expression levels of URAT1 and GLUT9 proteins in renal tissue and OXD protein in liver tissue of the model group were significantly increased， while the expression levels of renal OAT1 and ABCG2 were significantly decreased（P<0.01）. Compared with the model group， the expression levels of renal URAT1 and GLUT9 in the SFB granules group， Tongfengshu granules group and new formula group were significantly decreased， while the expression levels of renal OAT1 and ABCG2 were significantly increased， and the expression of XOD protein in liver tissue was significantly decreased（P<0.05， P<0.01）. Pathological analysis showed that focal infiltration of neutrophils， cell necrosis and nuclear fragmentation were observed in the liver tissue of the model group， sodium urate deposition crystals and tubular dilation appeared in renal tissue， synovial hyperplasia and inflammatory cell infiltration appeared in ankle joint. Compared with the model group， the abnormal degrees of liver， kidney and ankle joint tissue of rats in each treatment group were alleviated. ConclusionThe new formula of SFB replacing Plantaginis Semen has the same effect in the treatment of HUA combined with GA. This study proposes a new strategy to investigate the efficacy of new resources of Chinese medicine in a compound compatibility environment， which can provide a new demonstration for the research and development of new resources of Chinese medicine.

Objective To compare the changes of the medical animal resources(MAR)in Sichuan based on the data of the 3rd Chinese Materia Medica Resource Inventory(CMMRI,1983-1986)and the 4th CMMRI(2020-2022).Methods After field investigation and identification of the photos of the animals,the data of the MAR in Sichuan found in the 4th CMMRI were analyzed and compared with the data of 3rd CMMRI.Results 745 species of MAR were found in Sichuan during the 4th CMMRI,including 212 families and 468 genera.Compared with the 108 species found in 3rd CMMRI,the number of MAR in Sichuan had greatly increased,The Aves was found to have 243 species of MAR,which is the most plenty one among the 7 classes.There were 14 families which have more than 10 species of MAR.The family,Cyprinidae had 48 species of MAR.There were 33 common Chinese medicinal herbs and 3 genuine medicinal materials including Cordyceps sinensis,musk and Venenum bufonis were found in this investigation.The new distributions of Liangshan Cordyceps and Cordyceps gunnii were found in the investigation,and the new resources of Atypus heterothecus was found in Mountain Emei.There were 140 species of key protection of wild medicinal animals,including 44 species of animals under first-class protection and 96 species of animals under second-class protection.There were 230 species of animals which had important ecological,scientific,and social values,too.Conclusion Sichuan was plenty of medical animal resources and the protection of the forest musk deer,the Cordyceps sinensis and the pangolin was need to be strengthened.

Objective:To calculate the normalized plan complexity scores (nPCSs), assess their correlations with dose verification pass rates, and utilize nPCSs to compare the complexity of treatment plans for various anatomical parts.Methods:A retrospective analysis was conducted for 288 patients who underwent helical tomotherapy from January 2020 to November 2022, with 13 complexity parameters being extracted from the treatment plans. Principal components were extracted using principal component analysis (PCA), and those significantly correlated with the gamma pass rate based on the 3%/2 mm (global) criterion were selected to calculate the nPCSs. The correlations between nPCSs and gamma pass rates based on different criteria were assessed using multiple linear regression, and the complexity of treatment plans for different anatomical parts was compared.Results:The four principal components, PC1, PC2, PC3, PC4, accounted for 33.5%, 24.4%, 11.4%, and 9.2% of the total variance, respectively, and 78.48% in total. PC1, PC2, and PC3 were linearly correlated with the gamma pass rate based on the 3%/2 mm (global) criterion ( β = 0.34, 0.31, 0.62; P < 0.05). The nPCSs were linearly correlated with the gamma pass rates based on both the 3%/2 mm (global) and 1%/1 mm (global) criteria ( β = 5.36, 32.51, P < 0.001). The head and neck plans and abdominal-pelvic plans exhibited similar nPCS distributions, with no statistically significant difference ( P > 0.05). Conclusions:The nPCSs based on PCA can effectively evaluate the feasibility of treatment plans and compare the complexity of different treatment plans.

Objective:To calculate the normalized plan complexity scores (nPCSs), assess their correlations with dose verification pass rates, and utilize nPCSs to compare the complexity of treatment plans for various anatomical parts.Methods:A retrospective analysis was conducted for 288 patients who underwent helical tomotherapy from January 2020 to November 2022, with 13 complexity parameters being extracted from the treatment plans. Principal components were extracted using principal component analysis (PCA), and those significantly correlated with the gamma pass rate based on the 3%/2 mm (global) criterion were selected to calculate the nPCSs. The correlations between nPCSs and gamma pass rates based on different criteria were assessed using multiple linear regression, and the complexity of treatment plans for different anatomical parts was compared.Results:The four principal components, PC1, PC2, PC3, PC4, accounted for 33.5%, 24.4%, 11.4%, and 9.2% of the total variance, respectively, and 78.48% in total. PC1, PC2, and PC3 were linearly correlated with the gamma pass rate based on the 3%/2 mm (global) criterion ( β = 0.34, 0.31, 0.62; P < 0.05). The nPCSs were linearly correlated with the gamma pass rates based on both the 3%/2 mm (global) and 1%/1 mm (global) criteria ( β = 5.36, 32.51, P < 0.001). The head and neck plans and abdominal-pelvic plans exhibited similar nPCS distributions, with no statistically significant difference ( P > 0.05). Conclusions:The nPCSs based on PCA can effectively evaluate the feasibility of treatment plans and compare the complexity of different treatment plans.

Objective:To investigate effects of the autophagy inducer rapamycin and autophagy inhibitor 3-methyladenine on viral structures and biosynthesis of functional proteins in dorsal root ganglia in a guinea pig model of varicella-zoster virus (VZV) infection, and to explore their possible mechanisms.Methods:VZV was cultured and proliferated in human embryonic lung fibroblasts (HELFs) , and peripheral blood mononuclear cells (PBMCs) were isolated from guinea pigs. VZV-HELFs and PBMCs were co-cultured for 18-20 hours, and VZV-PBMCs were collected by centrifugation. Thirty-two guinea pigs were randomly and equally divided into 4 groups (8 mice in each group) : blank control group was injected with autologous PBMCs via the medial canthal venous plexus; autophagy inhibition group, autophagy induction group, and VZV infection group were intraperitoneally injected with 3 mg/kg 3-methyladenine solution, 0.5 mg/kg rapamycin solution, and the same volume of 0.9% NaCl solution respectively, followed 2 hours later by injections with 50 μl of VZV-PBMCs via the medial canthal venous plexus. Fourteen days later, the guinea pigs in each group were sacrificed, and dorsal root ganglion tissues were collected. The transmission electron microscope was used to observe the morphology of virus particles, as well as the morphology and number of autophagic vesicles, Western blot analysis was performed to determine the expression of VZV nucleocapsid protein (NCP) , immediate-early protein 62 (IE62) , and autophagy-related proteins Beclin-1 and p62, and immunohistochemical study to determine the expression of anti-VZV antibodies in VZV-infected dorsal root ganglia. Statistical analysis was carried out by using two-independent-sample t test, one-way analysis of variance, least significant difference- t test or Kruskal-Wallis H test. Results:Nucleocapsid-containing virions and scattered autophagosomes were seen in the dorsal root ganglia in the VZV infection group under the transmission electron microscope. The number of autophagic vesicles significantly differed among the blank control group, VZV infection group, autophagy induction group and autophagy inhibition group ( M[ Q1, Q3]: 0, 5[4, 6], 7[5, 9], 0, respectively; H = 135.60, P < 0.01) , and was significantly higher in the VZV infection group than in the blank control group and autophagy inhibition group (both P < 0.05) , as well as in the autophagy induction group than in the autophagy inhibition group ( P<0.05) , but there was no significant difference between the VZV infection group and autophagy induction group ( P>0.05) . Western blot analysis showed that the expression level of IE62 protein was significantly higher in the VZV infection group (1.49 ± 0.06) than in the blank control group (0.50 ± 0.09, t = 9.17, P < 0.05) ; the expression of anti-VZV antibodies was significantly lower in the autophagy inhibition group than in the autophagy induction group and VZV infection group ( t = 9.24, 7.78, respectively, both P < 0.01) , while there was no significant difference between the autophagy induction group and VZV infection group ( P > 0.05) . Conclusion:Autophagy occurred in the dorsal root ganglia of guinea pigs after VZV infection; the inhibition of autophagy could affect the structure of VZV and decrease the expression of VZV functional proteins in the dorsal root ganglia of guinea pigs.

Objective:To investigate the preliminary effects of sub-pectoral fascial breast augmentation and reconstruction.Methods:Six patients (10 breasts) of mammary dysplasia, mastatrophy, prophylactic mastectomy, unilateral breast defect with contralateral mastatrophy were included in the study from Oct. 2019 to July 2021. The mean patient age was 29 years (range, 20 to 35 years). All the textured and smooth cohesive gel implants were inserted under pectoralis major fascia by endoscopic-assisted, incisions were including axillary, around areola and inferior mammary fold approaches. The prophylactic mastectomy cases were performed nipple sparing mastectomy and sub-pectoral fascial breast restoration with implants. The fat and fascia tissue were well reserved to give a satisfying coverage of the appropriate implant. Negative pressure drainage and moulding dressing were used after the surgery.Results:Ten breast augmentation and restoration with a mean implant size of 200 cc (range, 180-300 cc). Less immediate postoperative pain and bleeding were reported. Patients were followed up for an average of 10 months (range, 6 to 21 months). There were no cases of major malpositions, double-bubble and bottom out symptoms. The overall outcome was with better symmetry and satisfaction with time passed by, and no secondary surgical procedures needed.Conclusions:Primary and satisfactory results have been obtained in subfascial breast augmentation and restoration for indications. With the endoscopic-assisted accurate pectoral fascial dissection, and well reserved soft tissue coverage, this technique could avoid the pectoralis major muscle ablation and keep the advantages of sub-glandular plane.

Objective:To investigate the application effect of reverse abdominal wall advancement flap in repairing chest wall soft tissue defect after breast tumor resection.Methods:From October 2020 to April 2021, the Department of Plastic Surgery and Breast Surgery of Hainan Cancer Hospital cooperated to repair the chest wall wounds of 4 female patients with unilateral giant breast tumors after primary lesion resection. Patients aged 40-63 years old, with an average of 51.5 years old. The size of the tumor estimated by physical examination was 7 cm × 6 cm-15 cm × 20 cm. The flaps were closely monitored after surgery, and complications were recorded. Local recurrence was followed-up.Results:The wound size of 4 patients after mastectomy was 16 cm×14 cm-20 cm×18 cm. Abdomen separation range reached anterior axillary line laterally and contralateral clavicle midline medially. Inferiorly, 1 case reached umbilical level, 1 case reached 2 cm below the umbilicus, and 2 cases reached 3 cm below the umbilicus. Three cases were diagnosed as breast lobulated tumors, and 1 case invasive lobular carcinoma. Among the 4 cases, 3 flaps survived completely and healed by first intention, while another flap healed under blister scab. There was no necrosis, infection, hematoma, seroma, or vascular crisis of the flap. All patients were satisfied. Three patients received radiotherapy and one received radiotherapy combined with oral chemotherapy. All patients were followed up for 3-6 months. No local recurrence was discovered. The abdomen was tighter than before operation, but no stiffness was found. There was no abdominal pain, abdominal wall bulging or abdominal hernia.Conclusions:Reverse abdominal wall advancedment flap was used to repair soft tissue defect of the chest wall after breast tumor resection. The operation was simple and fast, with no need for additional donor site incision. Quick recovery allowed shorter interval between surgery and next scheduled treatments.

Objective:To investigate the application effect of reverse abdominal wall advancement flap in repairing chest wall soft tissue defect after breast tumor resection.Methods:From October 2020 to April 2021, the Department of Plastic Surgery and Breast Surgery of Hainan Cancer Hospital cooperated to repair the chest wall wounds of 4 female patients with unilateral giant breast tumors after primary lesion resection. Patients aged 40-63 years old, with an average of 51.5 years old. The size of the tumor estimated by physical examination was 7 cm × 6 cm-15 cm × 20 cm. The flaps were closely monitored after surgery, and complications were recorded. Local recurrence was followed-up.Results:The wound size of 4 patients after mastectomy was 16 cm×14 cm-20 cm×18 cm. Abdomen separation range reached anterior axillary line laterally and contralateral clavicle midline medially. Inferiorly, 1 case reached umbilical level, 1 case reached 2 cm below the umbilicus, and 2 cases reached 3 cm below the umbilicus. Three cases were diagnosed as breast lobulated tumors, and 1 case invasive lobular carcinoma. Among the 4 cases, 3 flaps survived completely and healed by first intention, while another flap healed under blister scab. There was no necrosis, infection, hematoma, seroma, or vascular crisis of the flap. All patients were satisfied. Three patients received radiotherapy and one received radiotherapy combined with oral chemotherapy. All patients were followed up for 3-6 months. No local recurrence was discovered. The abdomen was tighter than before operation, but no stiffness was found. There was no abdominal pain, abdominal wall bulging or abdominal hernia.Conclusions:Reverse abdominal wall advancedment flap was used to repair soft tissue defect of the chest wall after breast tumor resection. The operation was simple and fast, with no need for additional donor site incision. Quick recovery allowed shorter interval between surgery and next scheduled treatments.