1.Mechanism of action of Sterculiae Lychnophorae Semen against PM2.5-induced acute lung injury based on network pharmacology,molecular docking and experimental validation
Fan ZHANG ; Yi-fan DU ; Xiao-shu DENG ; Zu-feng ZHANG ; Xian-lei HAN ; Wei TIAN ; Xiu-mei LI ; Mian CHEN ; Fei LIU ; Nan WANG
Chinese Pharmacological Bulletin 2025;41(12):2362-2369
Aim To investigate the anti-acute lung injury(ALI)effect of Sterculiae Lychnophorae Semen(SLS)and its mechanism.Methods The main ac-tive components of SLS and their core targets and path-ways of action against ALI were obtained by network pharmacology methods.Subsequently,molecular doc-king technology and in vitro cellular experiments were applied for validation.Results A total of 19 core tar-gets were obtained,including HSP90AA1,CASP3,TNF,MAPK8 and MAPK14.The mechanisms may in-volve signaling pathways such as cancer,PI3K/Akt and MAPK.Molecular docking confirmed that the key targets of SLS formed a better binding activity with the relevant active ingredients.The in vitro results showed that SLS was able to protect the PM2.5-contaminated BEAS-2B cells,inhibit their NO,IL-1β and TNF-αlevels,and reduce the expression of p-p38 MAPK and p-JNK proteins.Conclusions The study successfully predicts the active ingredients,targets and signaling pathways of SLS against ALI,and in vitro experiments demonstrate that SLS might protect BEAS-2B cells from PM2.5 stimulus-induced inflammation and apoptosis by inhibiting the over-activation of p38 MAPK and JNK signaling pathways.
2.Association of Body Mass Index with All-Cause Mortality and Cause-Specific Mortality in Rural China: 10-Year Follow-up of a Population-Based Multicenter Prospective Study.
Juan Juan HUANG ; Yuan Zhi DI ; Ling Yu SHEN ; Jian Guo LIANG ; Jiang DU ; Xue Fang CAO ; Wei Tao DUAN ; Ai Wei HE ; Jun LIANG ; Li Mei ZHU ; Zi Sen LIU ; Fang LIU ; Shu Min YANG ; Zu Hui XU ; Cheng CHEN ; Bin ZHANG ; Jiao Xia YAN ; Yan Chun LIANG ; Rong LIU ; Tao ZHU ; Hong Zhi LI ; Fei SHEN ; Bo Xuan FENG ; Yi Jun HE ; Zi Han LI ; Ya Qi ZHAO ; Tong Lei GUO ; Li Qiong BAI ; Wei LU ; Qi JIN ; Lei GAO ; He Nan XIN
Biomedical and Environmental Sciences 2025;38(10):1179-1193
OBJECTIVE:
This study aimed to explore the association between body mass index (BMI) and mortality based on the 10-year population-based multicenter prospective study.
METHODS:
A general population-based multicenter prospective study was conducted at four sites in rural China between 2013 and 2023. Multivariate Cox proportional hazards models and restricted cubic spline analyses were used to assess the association between BMI and mortality. Stratified analyses were performed based on the individual characteristics of the participants.
RESULTS:
Overall, 19,107 participants with a sum of 163,095 person-years were included and 1,910 participants died. The underweight (< 18.5 kg/m 2) presented an increase in all-cause mortality (adjusted hazards ratio [ aHR] = 2.00, 95% confidence interval [ CI]: 1.66-2.41), while overweight (≥ 24.0 to < 28.0 kg/m 2) and obesity (≥ 28.0 kg/m 2) presented a decrease with an aHR of 0.61 (95% CI: 0.52-0.73) and 0.51 (95% CI: 0.37-0.70), respectively. Overweight ( aHR = 0.76, 95% CI: 0.67-0.86) and mild obesity ( aHR = 0.72, 95% CI: 0.59-0.87) had a positive impact on mortality in people older than 60 years. All-cause mortality decreased rapidly until reaching a BMI of 25.7 kg/m 2 ( aHR = 0.95, 95% CI: 0.92-0.98) and increased slightly above that value, indicating a U-shaped association. The beneficial impact of being overweight on mortality was robust in most subgroups and sensitivity analyses.
CONCLUSION
This study provides additional evidence that overweight and mild obesity may be inversely related to the risk of death in individuals older than 60 years. Therefore, it is essential to consider age differences when formulating health and weight management strategies.
Humans
;
Body Mass Index
;
China/epidemiology*
;
Male
;
Female
;
Middle Aged
;
Prospective Studies
;
Rural Population/statistics & numerical data*
;
Aged
;
Follow-Up Studies
;
Adult
;
Mortality
;
Cause of Death
;
Obesity/mortality*
;
Overweight/mortality*
3.Analysis of laboratory outcomes in assisted reproductive technology for malignant tumor patients
Ran SHEN ; Wei ZHENG ; Ruowen ZU ; Chen YANG ; Bingnan REN ; Jiaheng LI ; Yanli LIU ; Jing LI ; Peixin LI ; Jingyi HAN ; Yichun GUAN
Chinese Journal of Reproduction and Contraception 2025;45(4):365-371
Objective:To investigate whether malignant tumors affect the laboratory outcomes of patients in their first controlled ovarian hyperstimulation (COH) cycle.Methods:This study was a retrospective case-control study that analyzed the clinical and laboratory data of patients who underwent fertility preservation before chemotherapy and radiotherapy due to malignant tumors, as well as patients with infertility caused by tubal factors who first underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) at the Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2020 to May 2024. Patients who underwent fertility preservation were designated as the research group, while patients who underwent assisted reproduction due to tubal factors during the same period were designated as control group. After 1∶3 propensity score matching (PSM), 40 patients were included in the research group and 118 patients were included in control group. The ovarian response, oocyte retrieval outcomes, and embryonic development after fertilization in the first COH cycle were compared between the two groups. Results:After PSM, the research group and control group showed statistically significant differences in the gonadotropin (Gn) starting dosage [225.00 (162.50, 300.00) U vs. 193.75 (150.00, 225.00) U, P=0.002], duration of Gn used [10.00 (8.00, 11.00) d vs. 12.00 (10.00, 13.00) d, P<0.001], and average estradiol levels on human chorionic gonadotropin trigger day [2 487.00 (1 461.25, 4 090.25) pmol/L vs. 10 738.50 (8 400.00, 16 507.25) pmol/L, P<0.001]. However, no statistically significant difference was found in the total dosages of Gn used between the two groups ( P>0.05). There were no significant differences between the groups in terms of the number of oocytes retrieved, the number of metaphase Ⅱ oocytes, two pronuclei (2PN) rate, 2PN cleavage rate, available embryo rate, high-quality embryo rate, blastocyst formation rate, and available blastocyst formation rate (all P>0.05). Conclusion:Compared with infertility patients with tubal factors, there is no significant difference in the laboratory outcomes of malignant tumor patients undergoing COH for fertility preservation prior to chemotherapy and radiation.
4.Impact of male body mass index on semen parameters and outcomes of artificial insemination by husband: a single-center retrospective cohort study
Jingyi HAN ; Chen YANG ; Ruowen ZU ; Peixin LI ; Ran SHEN ; Wei ZHENG ; Rusheng LIU ; Bingnan REN ; Yichun GUAN
Chinese Journal of Reproduction and Contraception 2025;45(6):600-606
Objective:To investigate the effects of male body mass index (BMI) on semen parameters and perinatal outcomes following artificial insemination by husband (AIH) treatment.Methods:A retrospective cohort study was conducted to analyze the clinical data of 5 053 patients underwent AIH treatment at the Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University, from January 2017 to February 2024. The study focused on factors such as male semen parameter abnormalities, male sexual dysfunction, female cervical factors, reproductive tract malformations, and unexplained infertility. Patients were classified into three groups based on male BMI: normal weight group (18.5-23.9 kg/m2, n=1 673), overweight group (24.0-27.9 kg/m2, n=2 078), and obese group (BMI≥28.0 kg/m2, n=1 302). The primary objective was to assess the differences in semen parameters and perinatal outcomes among the three groups. Multivariable logistic regression and linear regression analyses were applied to adjust for potential confounders that could influence semen parameters and perinatal outcomes. Results:Semen volume in the normal weight group and overweight group [4.00 (3.00, 5.50) mL, 4.00 (3.00, 5.50) mL] was higher than that in the obese group [4.00 (3.00, 5.00) mL], with a significant difference among the three groups ( P<0.001, a P<0.001). The total sperm count in the normal group and overweight group [207.60 (121.90, 341.75)×10 6, 211.80 (119.88, 334.83)×10 6] was higher than that in the obese group [188.40 (110.96, 323.41)×10 6], with a significant difference among the three groups ( P=0.007, a P<0.001). The total progressive sperm motility count in the normal group [88.18 (43.63, 163.80)×10 6] was higher than that in the obese group [75.30 (40.29, 147.86)×10 6], with a significant difference among the three groups ( P=0.001, a P<0.001). The percentage of forward motile sperm in the normal group [(45.37±17.16)%] was higher than that in the overweight group [(44.03±17.36)%] and the obese group [(43.80±17.21)%], with a significant difference compared among the three groups ( P=0.020, a P=0.016]. In terms of perinatal outcomes, after multivariate logistic regression analysis, only the overweight and obese groups had higher newborn birth weights [(3 389.53±472.65) g, (3 408.57±507.90) g] compared with the normal group [(3 271.32±532.02) g], with a significant difference among the three groups ( P=0.010, a P=0.009). Conclusion:Higher male BMI is associated with decreased semen quality and may increase newborn birth weight following AIH treatment.
5.Mediating role of serum β-hCG levels in the relationship between blastocyst quality and pregnancy outcomes in frozen-thawed single blastocyst transfer
Peixin LI ; Ruowen ZU ; Bingnan REN ; Jingyi HAN ; Wei ZHENG ; Chen YANG ; Yichun GUAN
Chinese Journal of Reproduction and Contraception 2025;45(6):582-590
Objective:To investigate the mediating role of serum β-human chorionic gonadotropin (hCG) levels on the relationship between embryo quality and pregnancy outcomes following single frozen-thawed blastocyst transfer 14 d post-transfer.Methods:This retrospective cohort study collected data from patients who underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) frozen-thawed single blastocyst transfer at the Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University between August 2017 and June 2021. Patients were grouped according to embryo quality into good-quality blastocyst group ( n=3 191) and available blastocyst group ( n=2 027). Differences in serum β-hCG levels and pregnancy outcomes at 14 d post-transfer were compared between the two groups. Mediation analysis and receiver operating characteristic (ROC) analysis were used to explore the mediating effect of β-hCG levels on the relationship between embryo quality and pregnancy outcomes and to evaluate the differences in the incidence of placental-related diseases between the two groups. Results:The good-quality blastocyst group had significantly higher serum β-hCG levels [1 177.0 (1.8, 2 278.5) U/L], clinical pregnancy rate [65.62% (2 094/3 191)], and live birth rate [52.55% (1 667/3 191)] compared with the available blastocyst group [54.4 (0.1, 1 453.5) U/L, P<0.001; 46.13% (935/2 027), P<0.001; 34.19% (693/2 027), P<0.001]. The early miscarriage rate in the good-quality group [13.47% (282/2 094)] was lower than that in the available blastocyst group [19.14% (179/935), P<0.001]. Serum β-hCG levels at 14 d post-transfer showed significant mediating effects on clinical pregnancy rate ( r=-0.126), live birth rate ( r=-0.122), and early miscarriage rate ( r=0.028) in both groups (all P<0.001). The cut-off values for β-hCG to predict live birth in the available and good-quality blastocyst groups were 366.9 U/L and 485.5 U/L, with positive predictive values of 76.28% (672/881) and 82.84% (1 628/1 965), respectively, and negative predictive values of 98.15% (1 114/1 135) and 96.14% (1 170/1 217). The cut-off values for predicting clinical pregnancy were 118.8 U/L and 226.5 U/L, with positive predictive values of 95.43% (919/963) and 98.45% (2 037/2 069), and negative predictive values of 99.72% (1 050/1 053) and 94.89% (1 059/1 116). The cut-off values for predicting early miscarriage were 1 337.0 U/L and 1 162.6 U/L, with positive predictive values of 32.75% (130/397) and 30.18% (150/497), and negative predictive values of 90.89% (489/538) and 91.73% (1 465/1 597). No differences were found in the incidence of placental-related diseases between the two groups (all P>0.05). Conclusion:This study indicates that both embryo quality and serum β-hCG levels at 14 d post-transfer significantly affect pregnancy outcomes. β-hCG levels play an important mediating role between embryo quality and pregnancy outcomes. ROC analysis demonstrates the good predictive efficacy of serum β-hCG levels for pregnancy outcomes, providing scientific evidence for optimizing embryo selection.
6.Analysis of laboratory outcomes in assisted reproductive technology for malignant tumor patients
Ran SHEN ; Wei ZHENG ; Ruowen ZU ; Chen YANG ; Bingnan REN ; Jiaheng LI ; Yanli LIU ; Jing LI ; Peixin LI ; Jingyi HAN ; Yichun GUAN
Chinese Journal of Reproduction and Contraception 2025;45(4):365-371
Objective:To investigate whether malignant tumors affect the laboratory outcomes of patients in their first controlled ovarian hyperstimulation (COH) cycle.Methods:This study was a retrospective case-control study that analyzed the clinical and laboratory data of patients who underwent fertility preservation before chemotherapy and radiotherapy due to malignant tumors, as well as patients with infertility caused by tubal factors who first underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) at the Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2020 to May 2024. Patients who underwent fertility preservation were designated as the research group, while patients who underwent assisted reproduction due to tubal factors during the same period were designated as control group. After 1∶3 propensity score matching (PSM), 40 patients were included in the research group and 118 patients were included in control group. The ovarian response, oocyte retrieval outcomes, and embryonic development after fertilization in the first COH cycle were compared between the two groups. Results:After PSM, the research group and control group showed statistically significant differences in the gonadotropin (Gn) starting dosage [225.00 (162.50, 300.00) U vs. 193.75 (150.00, 225.00) U, P=0.002], duration of Gn used [10.00 (8.00, 11.00) d vs. 12.00 (10.00, 13.00) d, P<0.001], and average estradiol levels on human chorionic gonadotropin trigger day [2 487.00 (1 461.25, 4 090.25) pmol/L vs. 10 738.50 (8 400.00, 16 507.25) pmol/L, P<0.001]. However, no statistically significant difference was found in the total dosages of Gn used between the two groups ( P>0.05). There were no significant differences between the groups in terms of the number of oocytes retrieved, the number of metaphase Ⅱ oocytes, two pronuclei (2PN) rate, 2PN cleavage rate, available embryo rate, high-quality embryo rate, blastocyst formation rate, and available blastocyst formation rate (all P>0.05). Conclusion:Compared with infertility patients with tubal factors, there is no significant difference in the laboratory outcomes of malignant tumor patients undergoing COH for fertility preservation prior to chemotherapy and radiation.
7.Impact of male body mass index on semen parameters and outcomes of artificial insemination by husband: a single-center retrospective cohort study
Jingyi HAN ; Chen YANG ; Ruowen ZU ; Peixin LI ; Ran SHEN ; Wei ZHENG ; Rusheng LIU ; Bingnan REN ; Yichun GUAN
Chinese Journal of Reproduction and Contraception 2025;45(6):600-606
Objective:To investigate the effects of male body mass index (BMI) on semen parameters and perinatal outcomes following artificial insemination by husband (AIH) treatment.Methods:A retrospective cohort study was conducted to analyze the clinical data of 5 053 patients underwent AIH treatment at the Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University, from January 2017 to February 2024. The study focused on factors such as male semen parameter abnormalities, male sexual dysfunction, female cervical factors, reproductive tract malformations, and unexplained infertility. Patients were classified into three groups based on male BMI: normal weight group (18.5-23.9 kg/m2, n=1 673), overweight group (24.0-27.9 kg/m2, n=2 078), and obese group (BMI≥28.0 kg/m2, n=1 302). The primary objective was to assess the differences in semen parameters and perinatal outcomes among the three groups. Multivariable logistic regression and linear regression analyses were applied to adjust for potential confounders that could influence semen parameters and perinatal outcomes. Results:Semen volume in the normal weight group and overweight group [4.00 (3.00, 5.50) mL, 4.00 (3.00, 5.50) mL] was higher than that in the obese group [4.00 (3.00, 5.00) mL], with a significant difference among the three groups ( P<0.001, a P<0.001). The total sperm count in the normal group and overweight group [207.60 (121.90, 341.75)×10 6, 211.80 (119.88, 334.83)×10 6] was higher than that in the obese group [188.40 (110.96, 323.41)×10 6], with a significant difference among the three groups ( P=0.007, a P<0.001). The total progressive sperm motility count in the normal group [88.18 (43.63, 163.80)×10 6] was higher than that in the obese group [75.30 (40.29, 147.86)×10 6], with a significant difference among the three groups ( P=0.001, a P<0.001). The percentage of forward motile sperm in the normal group [(45.37±17.16)%] was higher than that in the overweight group [(44.03±17.36)%] and the obese group [(43.80±17.21)%], with a significant difference compared among the three groups ( P=0.020, a P=0.016]. In terms of perinatal outcomes, after multivariate logistic regression analysis, only the overweight and obese groups had higher newborn birth weights [(3 389.53±472.65) g, (3 408.57±507.90) g] compared with the normal group [(3 271.32±532.02) g], with a significant difference among the three groups ( P=0.010, a P=0.009). Conclusion:Higher male BMI is associated with decreased semen quality and may increase newborn birth weight following AIH treatment.
8.Mediating role of serum β-hCG levels in the relationship between blastocyst quality and pregnancy outcomes in frozen-thawed single blastocyst transfer
Peixin LI ; Ruowen ZU ; Bingnan REN ; Jingyi HAN ; Wei ZHENG ; Chen YANG ; Yichun GUAN
Chinese Journal of Reproduction and Contraception 2025;45(6):582-590
Objective:To investigate the mediating role of serum β-human chorionic gonadotropin (hCG) levels on the relationship between embryo quality and pregnancy outcomes following single frozen-thawed blastocyst transfer 14 d post-transfer.Methods:This retrospective cohort study collected data from patients who underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) frozen-thawed single blastocyst transfer at the Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University between August 2017 and June 2021. Patients were grouped according to embryo quality into good-quality blastocyst group ( n=3 191) and available blastocyst group ( n=2 027). Differences in serum β-hCG levels and pregnancy outcomes at 14 d post-transfer were compared between the two groups. Mediation analysis and receiver operating characteristic (ROC) analysis were used to explore the mediating effect of β-hCG levels on the relationship between embryo quality and pregnancy outcomes and to evaluate the differences in the incidence of placental-related diseases between the two groups. Results:The good-quality blastocyst group had significantly higher serum β-hCG levels [1 177.0 (1.8, 2 278.5) U/L], clinical pregnancy rate [65.62% (2 094/3 191)], and live birth rate [52.55% (1 667/3 191)] compared with the available blastocyst group [54.4 (0.1, 1 453.5) U/L, P<0.001; 46.13% (935/2 027), P<0.001; 34.19% (693/2 027), P<0.001]. The early miscarriage rate in the good-quality group [13.47% (282/2 094)] was lower than that in the available blastocyst group [19.14% (179/935), P<0.001]. Serum β-hCG levels at 14 d post-transfer showed significant mediating effects on clinical pregnancy rate ( r=-0.126), live birth rate ( r=-0.122), and early miscarriage rate ( r=0.028) in both groups (all P<0.001). The cut-off values for β-hCG to predict live birth in the available and good-quality blastocyst groups were 366.9 U/L and 485.5 U/L, with positive predictive values of 76.28% (672/881) and 82.84% (1 628/1 965), respectively, and negative predictive values of 98.15% (1 114/1 135) and 96.14% (1 170/1 217). The cut-off values for predicting clinical pregnancy were 118.8 U/L and 226.5 U/L, with positive predictive values of 95.43% (919/963) and 98.45% (2 037/2 069), and negative predictive values of 99.72% (1 050/1 053) and 94.89% (1 059/1 116). The cut-off values for predicting early miscarriage were 1 337.0 U/L and 1 162.6 U/L, with positive predictive values of 32.75% (130/397) and 30.18% (150/497), and negative predictive values of 90.89% (489/538) and 91.73% (1 465/1 597). No differences were found in the incidence of placental-related diseases between the two groups (all P>0.05). Conclusion:This study indicates that both embryo quality and serum β-hCG levels at 14 d post-transfer significantly affect pregnancy outcomes. β-hCG levels play an important mediating role between embryo quality and pregnancy outcomes. ROC analysis demonstrates the good predictive efficacy of serum β-hCG levels for pregnancy outcomes, providing scientific evidence for optimizing embryo selection.
9.Mechanism of action of Sterculiae Lychnophorae Semen against PM2.5-induced acute lung injury based on network pharmacology,molecular docking and experimental validation
Fan ZHANG ; Yi-fan DU ; Xiao-shu DENG ; Zu-feng ZHANG ; Xian-lei HAN ; Wei TIAN ; Xiu-mei LI ; Mian CHEN ; Fei LIU ; Nan WANG
Chinese Pharmacological Bulletin 2025;41(12):2362-2369
Aim To investigate the anti-acute lung injury(ALI)effect of Sterculiae Lychnophorae Semen(SLS)and its mechanism.Methods The main ac-tive components of SLS and their core targets and path-ways of action against ALI were obtained by network pharmacology methods.Subsequently,molecular doc-king technology and in vitro cellular experiments were applied for validation.Results A total of 19 core tar-gets were obtained,including HSP90AA1,CASP3,TNF,MAPK8 and MAPK14.The mechanisms may in-volve signaling pathways such as cancer,PI3K/Akt and MAPK.Molecular docking confirmed that the key targets of SLS formed a better binding activity with the relevant active ingredients.The in vitro results showed that SLS was able to protect the PM2.5-contaminated BEAS-2B cells,inhibit their NO,IL-1β and TNF-αlevels,and reduce the expression of p-p38 MAPK and p-JNK proteins.Conclusions The study successfully predicts the active ingredients,targets and signaling pathways of SLS against ALI,and in vitro experiments demonstrate that SLS might protect BEAS-2B cells from PM2.5 stimulus-induced inflammation and apoptosis by inhibiting the over-activation of p38 MAPK and JNK signaling pathways.
10.Comparison of the pharmacokinetics of gatifloxacin ophthalmic gel in rabbit ocular tissues with different dosing frequencies
Xiaotong WANG ; Xin ZHAO ; Wei ZU ; Xu SHEN ; Shichao CHEN ; Qiang YANG
Chinese Journal of Experimental Ophthalmology 2024;42(7):621-628
Objective:To establish a method for determining the concentration of gatifloxacin in rabbit ocular tissue and compare the ocular pharmacokinetics of 0.3% gatifloxacin eye gel after a single and multiple topical instillations in rabbits.Methods:Ninety-four healthy New Zealand rabbits were selected.Ten rabbits were randomly selected without any treatment for blank tissue collection, and the remaining 84 rabbits were randomly divided into a single-dose group (36 rabbits) and a multiple-dose group (48 rabbits) equally between males and females using a random number table.The left eye was taken as the experimental eye.The single-dose group was given one drop of 0.3% gatifloxacin eye gel into the left eyes, and the rabbits were divided evenly into six subgroups.In each subgroup, tear specimens and blood specimens were collected at 0.5, 1, 3, 5, 7, 10 hours after gel application, then cardiac blood samples were taken, after which animals were sacrificed immediately to collect ocular tissue including aqueous humor, conjunctiva, cornea, sclera, iris-ciliary body, lens, vitreous body, retina, and choroid.The multiple-dose group was given 1 drop of gatifloxacin ophthalmic gel in the left eye three times a day.At 0.5 hour after the first administration days 4 and 6, and 0.5, 1, 3, 5, 7, and 10 hours after the first administration on day 7, the cardiac blood sampling and ocular tissue collection were performed.The methanol precipitation protein method was used to pretreat samples, and the concentration of gatifloxacin in rabbit plasma and eye tissue was measured and calculated by high-performance liquid chromatography-tandem mass spectrometry method to obtain pharmacokinetic-related parameters such as peak concentration (C max), peak time (T max), and area under curve (AUC).The mobile phase was a methanol-0.1% acetic acid aqueous solution (volume ratio=70∶30), and a positive ion multiple reaction detection mode was used.Ciprofloxacin was used as the internal standard, the selectivity, standard curve and lower limit of quantification, accuracy and precision, extraction recovery rate, matrix effect, and stability of the method were validated in accordance with the 9012 Guidelines for Validation of Quantitative Analysis Methods for Biological Samples in Chinese Pharmacopoeia ( 2020 edition).Combined with the minimum inhibitory concentration (MIC 90) of gatifloxacin on common ocular infectious bacteria, C max/MIC 90 and AUC/MIC 90 were calculated.The study protocol was reviewed and approved by the Animal Ethics Committee of Shenyang Xingqi Pharmaceutical Co., Ltd.(No.XQ-2016-011). Results:Gatifloxacin has a good linear relationship in various eye tissues and plasma.The between-run precision in corneal tissue is within the range of -1.5%-6.0%, and the daytime precision was not greater than 15%.The extraction recovery rate in corneal tissue ranged from 92.0% to 94.8%, and the precision of the matrix effect at low, medium, and high concentrations calculated by internal standard normalization was not greater than 3.3%.After a single topical instillation, gatifloxacin reached a high concentration in anterior and posterior segment ocular tissues and its distribution ranked in order from the highest to the lowest by AUC 0-t as follows, tears, cornea, conjunctiva, iris-ciliary body, sclera, aqueous humor, choroid, retina, lens and vitreous body, with the C max of 94.90 μg/g, 7.34 μg/g, 3.65 μg/g, 1.81 μg/g, 1.75 μg/g, 1.31 μg/ml, 0.86 μg/g, 0.53 μg/g, 0.13 μg/g and 0.07 μg/ml, respectively.T max was 1 hour in all ocular tissues except in the lens, choroid, and vitreous body fluid, where T max was 0.5 hour.There was no significant difference among the concentrations of gatifloxacin in ocular tissues at 0.5 hour on days 4, 6 and 7 after multiple dosing ( P>0.05), and the AUC 0-t in the cornea, conjunctiva, and sclera was approximately 2.04, 2.12, and 2.32 times that of the single dosing.The concentration of gatifloxacin released into the systemic circulation after single and multiple dosing was less than 25.00 ng/ml.For both Staphylococcus aureus and Staphylococcus epidermidis, pharmacokinetic/pharmacodynamics in the conjunctiva, cornea, sclera, iris-ciliary body, aqueous humor, and choroid were satisfied with C max/MIC 90≥10 and AUC/MIC 90≥30 after continuous administration of gatifloxacin ophthalmic gel. Conclusions:A rapid and sensitive method for measuring gatifloxacin concentration in ocular tissues is successfully constructed.Gatifloxacin ophthalmic gel administered three times a day for three days can achieve stable concentrations in ocular tissues, and the concentration of gatifloxacin in ocular tissues is increased compared with a single dose.Effective treatment of common bacterial infections of the conjunctiva, cornea, sclera, and iris-ciliary body can be achieved with topical application of gatifloxacin ophthalmic gel.

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