ObjectiveTo investigate the effect of the herb pair Cuscutae Semen-Lycii Fructus on oxidative stress-induced blood-testis barrier dysfunction and spermatogenesis in the rat model of oligoasthenozoospermia （OAS） and decipher the mechanism based on the silent information regulator 1 （SIRT1）/nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway. MethodsThirty-five male SD rats were randomized into a blank group （n=7） and a modeling group （n=28）. The OAS model was established by gavage of hydrocortisone aqueous solution combined with single factor electrical stimulation. The modeled rats were randomly assigned into the following groups： model， Cuscutae Semen-Lycii Fructus granules （3.2 g·kg^-1）， Cuscutae Semen-Lycii Fructus total flavonoids （0.34 g·kg^-1）， and L-carnitine （0.38 g·kg^-1）， and treated for 4 weeks. The sperm quality of rats was assessed by an automatic sperm analyzer. The levels of superoxide dismutase （SOD）， malondialdehyde （MAD）， and glutathione peroxidase （GSH-Px） in the testicular tissue were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the pathological changes in the testicular tissue and score the spermatogenic function. Transmission electron microscopy was employed to observe the ultrastructural changes of Sertoli cells. Western blot and Real-time PCR were employed to determine the protein and mRNA levels， respectively， of SIRT1， Nrf2， Occludin， zonula occludens-1 （ZO-1）， connexin 43 （CX43）， and β-catenin. ResultsCompared with the blank group， the model group showed decreased total sperm count and motility （P<0.05， P<0.01）， obvious damage in the testicular tissue and blood-testis barrier structure， reduced score of spermatogenic function （P<0.01）， declined levels of GSH-Px and SOD in the testicular tissue （P<0.05）， elevated level of MDA， and down-regulated protein levels of SIRT1， Nrf2， ZO-1， CX43， β-catenin， and occludin （P<0.05， P<0.01） and mRNA levels of SIRT1， Nrf2， ZO-1， CX43， and β-catenin in the testicular tissue （P<0.05， P<0.01）. After treatment， the testicular tissue， blood-testis barrier structure， and score of spermatogenic function （P<0.01） were improved in the Cuscutae Semen-Lycii Fructus granules group， Cuscutae Semen-Lycii Fructus total flavonoids group， and L-carnitine group. Compared with the model group， the treatment groups presented lowered levels of GSH-Px and SOD （P<0.05， P<0.01）， and the Cuscutae Semen-Lycii Fructus granule group showed a decline in MDA level. The protein and mRNA levels of SIRT1， Nrf2， ZO-1， CX43， β-catenin， and occludin were up-regulated in the Cuscutae Semen-Lycii Fructus granules group and total flavonoids group （P<0.05， P<0.01）. ConclusionThe herb pair Cuscutae Semen-Lycii Fructus can regulate the SIRT1/Nrf2 pathway to inhibit oxidative stress and alleviate the blood-testis barrier damage， thereby improving the spermatogenic function in the rat model of OAS. Total flavonoids may be the material basis for the therapeutic effect of Cuscutae Semen-Lycii Fructus.

ObjectiveTo investigate the effect of the herb pair Cuscutae Semen-Lycii Fructus on oxidative stress-induced blood-testis barrier dysfunction and spermatogenesis in the rat model of oligoasthenozoospermia （OAS） and decipher the mechanism based on the silent information regulator 1 （SIRT1）/nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway. MethodsThirty-five male SD rats were randomized into a blank group （n=7） and a modeling group （n=28）. The OAS model was established by gavage of hydrocortisone aqueous solution combined with single factor electrical stimulation. The modeled rats were randomly assigned into the following groups： model， Cuscutae Semen-Lycii Fructus granules （3.2 g·kg^-1）， Cuscutae Semen-Lycii Fructus total flavonoids （0.34 g·kg^-1）， and L-carnitine （0.38 g·kg^-1）， and treated for 4 weeks. The sperm quality of rats was assessed by an automatic sperm analyzer. The levels of superoxide dismutase （SOD）， malondialdehyde （MAD）， and glutathione peroxidase （GSH-Px） in the testicular tissue were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the pathological changes in the testicular tissue and score the spermatogenic function. Transmission electron microscopy was employed to observe the ultrastructural changes of Sertoli cells. Western blot and Real-time PCR were employed to determine the protein and mRNA levels， respectively， of SIRT1， Nrf2， Occludin， zonula occludens-1 （ZO-1）， connexin 43 （CX43）， and β-catenin. ResultsCompared with the blank group， the model group showed decreased total sperm count and motility （P<0.05， P<0.01）， obvious damage in the testicular tissue and blood-testis barrier structure， reduced score of spermatogenic function （P<0.01）， declined levels of GSH-Px and SOD in the testicular tissue （P<0.05）， elevated level of MDA， and down-regulated protein levels of SIRT1， Nrf2， ZO-1， CX43， β-catenin， and occludin （P<0.05， P<0.01） and mRNA levels of SIRT1， Nrf2， ZO-1， CX43， and β-catenin in the testicular tissue （P<0.05， P<0.01）. After treatment， the testicular tissue， blood-testis barrier structure， and score of spermatogenic function （P<0.01） were improved in the Cuscutae Semen-Lycii Fructus granules group， Cuscutae Semen-Lycii Fructus total flavonoids group， and L-carnitine group. Compared with the model group， the treatment groups presented lowered levels of GSH-Px and SOD （P<0.05， P<0.01）， and the Cuscutae Semen-Lycii Fructus granule group showed a decline in MDA level. The protein and mRNA levels of SIRT1， Nrf2， ZO-1， CX43， β-catenin， and occludin were up-regulated in the Cuscutae Semen-Lycii Fructus granules group and total flavonoids group （P<0.05， P<0.01）. ConclusionThe herb pair Cuscutae Semen-Lycii Fructus can regulate the SIRT1/Nrf2 pathway to inhibit oxidative stress and alleviate the blood-testis barrier damage， thereby improving the spermatogenic function in the rat model of OAS. Total flavonoids may be the material basis for the therapeutic effect of Cuscutae Semen-Lycii Fructus.

This paper summarized Professor ZHANG Farong's experience in the staged treatment of hyperthyroi-dism with triangular medicine. It is considered that "pathogenic heat consuming and damaging qi-yin" is the key pathogenesis of hyperthyroidism. In the early stage， liver constraint transforms into fire， and then pathogenic heat becomes exuberant inside， consuming qi and damaging yin. In the middle stage， the fire and heat are intense， and phlegm and stasis are binded， when qi and yin are initially depleted. In the late stage， both qi and yin are deficient， and as the disease lasts for a long time， the kidneys are affected， resulting in liver and kidney depletion. For treatment， it is suggested to put focus on rectifying healthy qi and dispelling pathogen， protecting qi and yin， and treating the disease by stages based on syndrome differentiation， and eight groups of triangular medicines have been summarized according to the characteristics of pathogenesis in different periods. In the early stage， the combination of Chaihu （Radix Bupleuri）， Xiangfu （Rhizoma Cyperi） and Yujin （Radix Curcumae） is used to soothe the liver and resolve constraint， and the combination of Longdancao （Radix et Rhizoma Gentianae）， Huangqin （Radix Scutellariae） and Zhizi （Fructus Gardeniae） is recommended to clear liver and drain fire. In the middle stage， the combination of Xiakucao （Spica Prunellae）， Baihua Sheshecao （Herba Hedyotis Diffusae） and Banzhilian （Herba Scutellariae Barbatae） is used to clear heat and dissipate masses， and the combination of Huangyaozi （Rhizoma Dioscoreae bulbiferae）， Maozhuacao （Ranunculus Ternatus） and Shancigu （Asarum Sagittarioides） can dissolve phlegm and dissipate masses， and the combination of Danshen （Radix et Rhizoma Salviae Miltiorrhizae）， Xuanshen （Radix Scrophulariae） and Kushen （Radix Sophorae Flavescentis） can invigorate blood and eliminate goiter. In the late stage， the combination of Dangshen （Radix Codonopsis）， Baizhu （Rhizoma Atractylodis Macrocephalae） and Yiyiren （Semen Coicis） can fortify spleen and replenish qi， and the combination of Maidong （Radix Ophiopogonis）， Beishashen （Radix Glehniae） and Wuweizi （Fructus Schisandrae Chinensis） is used to nourish yin and calm heart， while the combination of Huangqi （Radix Astragali）， Huangjing （Rhizoma Polygonati） and Diyu （Radix Sanguisorbae） is used to nourish yin and replenish kidneys.

Vitiligo is a common acquired depigmentation disorder characterized by the loss of melanocytes in the skin,hair,and mucous membranes.Its characteristics of difficulty in re-pigmentation and easy recurrence are great challenges for vitiligo treat-ment.At present,vitiligo has been clearly classified as an autoimmune disease with a complex pathogenesis involving genetic and envi-ronmental factors,oxidative stress and autoimmune responses that synergistically promote the loss of melanocytes.With the in-depth understanding of the pathogenesis of vitiligo,a large number of clinical trials have been carried out for targeted therapy of key small molecules,cell subsets,and immune-related signaling pathways in vitiligo.Targeted therapy will become the mainstream in the treat-ment of vitiligo in the future.Research progress of targeted therapy for tissue resident memory T cells(TRM)pathway,JAK-STAT pathway,and WNT/β-catenin pathway in vitiligo.

Objective:To evaluate protective effects of isorhamnetin on mitochondrial structure and function in HaCaT cells under oxidative stress.Methods:HaCaT cells served as the research object, and were divided into 4 groups: control group receiving conventional culture, isorhamnetin group treated with 60 μmol/L isorhamnetin, H 2O 2 group treated with 600 μmol/L H 2O 2, and isorhamnetin + H 2O 2 group pretreated with 60 μmol/L isorhamnetin for 12 hours followed by medium replacement and 12-hour treatment with 600 μmol/L H 2O 2. Flow cytometry was performed to detect cellular reactive oxygen species (ROS) levels, transmission electron microscopy to observe mitochondrial ultrastructure, confocal fluorescence microscopy to evaluate mitochondrial membrane potential, real-time fluorescence-based quantitative PCR (qRT-PCR) to determine the mitochondrial DNA copy number, and adenosine triphosphate (ATP) assay kit was used to determine the mitochondrial ATP content. One-way analysis of variance was used for comparisons among multiple groups, and least significant difference- t test for multiple comparisons. Results:Oxidative stress was provoked in HaCaT cells after the treatment with H 2O 2. Compared with the control group, the H 2O 2 group showed significantly increased ROS levels (10 725.0 ± 845.8 vs. 1 708.0 ± 69.4, t = 18.40, P < 0.001), but significantly decreased fluorescence intensity of mitochondrial membrane potential, mitochondrial ATP content, and expression of ND-1 (a characteristic gene of mitochondrial DNA) ( t = 4.58, 4.48, 6.11, P = 0.010, 0.010, 0.003, respectively). After the pretreatment with isorhamnetin followed by H 2O 2 treatment, the isorhamnetin+ H 2O 2 group showed significantly decreased ROS levels (7 640.0 ± 922.7) compared with the H 2O 2 group ( t = 4.27, P = 0.013), but significantly increased fluorescence intensity of mitochondrial membrane potential, mitochondrial ATP content, and expression of ND-1 compared with the H 2O 2 group ( t = 4.59, 4.58, 5.61, P = 0.010, 0.010, 0.005, respectively). Under the electron microscope, the mitochondrial structure was clearer and more complete in the isorhamnetin+ H 2O 2 group than in the H 2O 2 group; there was slight or no swelling of mitochondrial cristae, and no vacuolization of mitochondria in the isorhamnetin+ H 2O 2 group; in addition, autophagosomes engulfing damaged mitochondria were observed in the isorhamnetin+ H 2O 2 group. Conclusion:Isorhamnetin may reduce ROS levels by inducing autophagy, and has a protective effect against the H 2O 2-induced mitochondrial structural and functional damage in HaCaT cells.

Objective To investigate the value of serum chitinase-3-like protein 1 (CHI3L1) in predicting the risk of decompensation events in patients with liver cirrhosis, since prediction of decompensation events and adoption of active preventive measures are the key to improving the survival time of patients with liver cirrhosis. Methods A case-control study was conducted for 305 patients with liver cirrhosis who were diagnosed and treated in Tianjin Second People's Hospital from January 2019 to May 2021, among whom there were 200 patients with compensated liver cirrhosis and 105 patients with decompensated liver cirrhosis at baseline. According to whether decompensation events occurred within 1 year, the 305 patients with liver cirrhosis were divided into decompensation group with 79 patients and non-decompensation group with 226 patients; according to whether decompensation events occurred for the first time within 1 year, the 200 patients with compensated liver cirrhosis were divided into first-time decompensation group with 43 patients and non-first-time decompensation group with 157 patients. The independent samples t -test or the Mann-Whitney U test was used for comparison of normally distributed continuous data between groups, and the Wilcoxon rank-sum test or the chi-square test was used for comparison of categorical data between groups. The binary logistic regression analysis was used to investigate the association between each variable and decompensation events; the receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC) were used to investigate the value of each variable in predicting decompensation events, and the maximum value of Youden index was used to determine the optimal cut-off value. Results The patients who experienced decompensation events within 1 year had a significantly higher baseline serum level of CHI3L1 than those who did not experience such events [243.00 (136.00-372.00) ng/mL vs 117.50 (67.75-205.25) ng/mL, U =4720.500, P < 0.001], and the patients who experienced decompensation events for the first time within 1 year had a significantly higher baseline serum level of CHI3L1 than those who did not experience such events [227.98 (110.00-314.00) ng/mL vs 90.00 (58.00-168.50) ng/mL, U =1 681.500, P < 0.001]. Patients with cirrhosis with higher baseline CHI3L1 levels had an increased risk of decompensation events within 1 year ( OR =1.004, 95% CI : 1.002-1.006, P < 0.001); Patients with compensated cirrhosis with higher baseline serum CHI3L1 levels had an increased risk of first decompensated event within 1 year ( OR =1.006, 95% CI : 1.003-1.008, P < 0.001). The baseline serum level of CHI3L1 had an AUC of 0.751 in predicting the risk of first-time decompensation events, with a sensitivity of 90.7% and a specificity of 55.4% at the optimal cut-off value of 95.5 ng/mL. The predictive model based on the combination of serum CHI3L1 level and Child-Pugh class had an AUC of 0.809, with a sensitivity of 72.1% and a specificity of 77.1% at the maximum value of Youden index. Conclusion Serum CHI3L1 level can be used as an effective predictive factor for the risk of first-time decompensation events in patients with compensated liver cirrhosis, and its combination with Child-Pugh class shows a higher predictive value.

Objective:To evaluate the effectiveness of prevention and control of coal-burning-borne endemic fluorosis in Sichuan Province, and to provide basis for further development of targeted prevention and control measures.Methods:According to the requirements of the Implementation Plan for the Elimination and Evaluation of Coal-burning-borne Endemic Fluorosis in Sichuan Province, from 2018 to 2021, the assessment of the elimination of coal-burning-borne endemic fluorosis was carried out in all the endemic fluorosis villages of 23 counties (cities, districts) in Sichuan Province. In each village, 30 households were selected to be investigated the use of improved stoves, the drying of corn and pepper for human consumption, and the prevalence of dental fluorosis among all children aged 8 - 12 years old in the village.Results:A total of 222 242 households were surveyed in 1 640 villages, 196 towns, 23 counties (cities, districts) in Sichuan Province. Among them, the rate of qualified improved stoves was 99.99% (222 238/222 242), the correct use rate of qualified improved stoves was 99.90% (212 026/222 238), and the correct drying rate of corn and pepper for human consumption was 99.96% (212 630/212 721). A total of 135 125 children aged 8 to 12 years old were examined, and 5 927 cases of dental fluorosis were detected. The prevalence of dental fluorosis in children was 4.39%. The grading of dental fluorosis was mainly very mild and mild, which were 2.71% (3 656/135 125) and 1.32% (1 783/135 125), respectively. In the county-level self-assessment of 1 640 villages, 1 621 villages had reached the elimination standard, with the elimination rate of 98.84%. Taking counties as a unit, the elimination rate of diseased villages ranged from 96.69% to 100.00%, and 23 diseased counties (cities, districts) had reached the elimination standard. In the provincial-level review results of 174 villages with endemic fluorosis, 174 diseased villages had reached the elimination standard, with the consistency rate of 98.85% (172/174) with the county-level self-assessment results. Among them, except for Gulin County, the consistency rate of self-assessment results was 7/9, the consistency rate of other counties (cities, districts) was 100.00%.Conclusions:The prevention and control of coal-burning-borne endemic fluorosis in Sichuan Province has achieved remarkable results, and all the diseased counties (cities, districts) have reached the elimination standard. In the future, we should continue to take active prevention and control measures to ensure that the elimination status is maintained.

Objective:To explore the influence factors of neurodevelopmental disorders in children with SCN8A-related early-onset epilepsy through analyzing their clinical characteristics and following up their neurodeve-lopmental status. Methods:A retrospective analysis was carried out on 21 children (13 males and 8 females, the age ranged from 4 months to 8 years, average 31.6 months)with SCN8A-related early-onset epilepsy treated in Guangzhou Women and Children′s Medical Center and Kunming Children′s Hospital between January 2017 and February 2021.All patients underwent whole-exome sequencing and Sanger sequencing.The pathogenicity was estimated according to the American College of Medical Genetics and Genomics guidelines.The clinical data of all patients were also collected, including the age of onset of the disease, forms of seizures, seizure frequency, neurological development at onset, electroencephalogram (EEG) and brain magnetic resonance imaging (MRI). Besides, the patients were followed up to acquire the effect of sodium channel blockers after the onset of seizures, the process or improvement of neurodeve-lopment, EEG evaluation and neurodevelopmental outcomes.Patients were grouped based on data analysis results.The Fisher′s exact test was conducted to measure the effect of various factors on the neurodevelopmental process and outcome, and corresponding coe-fficients were calculated. Results:The average onset age of 21 patients was 0-9 months.The follow-up duration was 4 months-8 years.Three cases died.Sixteen cases (76.2%) had early infantile epileptic encephalopathy (EIEE), 5 cases (23.8%) had epilepsy without encephalopathy, and 1 case had benign infantile epilepsy.Fourteen cases (66.7%) belonged to drug resistant epilepsy.Only one child showed normal neurodevelopment.Eleven children showed delayed neurodevelopment, but improvement was observed.Nine children were retrogressed and stagnated in terms of neurodevelopment.Small age at onset ( Fisher=9.517, P=0.020, r=0.571), high seizure frequency ( Fisher=10.512, P=0.003, r=0.572), EEG background ( Fisher=10.512, P=0.003, r=0.572), epileptic discharges ( Fisher=8.288, P=0.008, r=0.542), and EEG changes before and after treatment ( Fisher=10.437, P=0.009, r=0.586) were important factors affecting the neurodevelopmental process.Neurodevelopmental outcome was normal in only 1 case, 1 child belonged to mild mental retardation (MR), 7 children belonged to moderate MR, 3 children belonged to severe MR, and 9 children belonged to profound MR.Statistical analysis indicated that the clinical phenotype ( Fisher=10.059, P=0.004, r=0.739) and drug resistance ( Fisher=13.706, P=0.001, r=0.640) were significantly correlated with neurodevelopmental outcomes.However, the forms of seizures, EEG findings at onset and mutation sites were not related to neurodevelopmental disorders. Conclusions:Most children with SCN8A-related early-onset epilepsy are accompanied with neurodevelopmental retardation of varying degrees.Epileptic encephalopathy and poor response to drug treatment will lead to severe neurodevelopmental disorders.

Objective:To investigate the protective effect of isorhamnetin on oxidative stress injury of HaCaT cells induced by H 2O 2. Methods:HaCaT cells were cultured in vitro and treated with different concentrations of H 2O 2 (300, 600, 900, 1 200 μmol/L) for 12 h. Cell proliferation activity was detected by cell counting kit-8 (CCK-8) assay; SOD activity was detected by superoxide dismutase (SOD) kit and malondialdehyde (MDA) content was detected by MDA assay. The oxidative stress model was established by the selection of suitable H 2O 2 concentration. HaCaT cells were pretreated with isorhamnetin at different concentrations for 12 h, and cell survival rate was detected by CCK-8 method to determine the safe concentration of isorhamnetin for subsequent experiments. HaCaT cells were pretreated with safe concentration of isorhamnetin for 12 h, and H 2O 2 was used to interfere with HaCaT cells for 12 h. Cell proliferation activity, SOD activity and MDA content were detected. Results:With the increase of H 2O 2 concentration, the cell survival rate decreased gradually, the SOD activity decreased gradually and MDA content increased gradually. Compared with the control group, the survival rate of 600, 900 and 1 200 μmol/L H 2O 2 groups was statistically significant ( P<0.05); The SOD activity and MDA content of H 2O 2 groups (300, 600, 900, 1 200 μmol/L) were significantly different from those of the control group ( P<0.05). The oxidative stress model of HaCaT cells was established by 600 μmol/L H 2O 2. HaCaT cells treated with 20, 40, 60, 80 and 100 μmol/L isorhamnetin for 12 h showed no cytotoxic effect. 20, 40 and 60 μmol/L isorhamnetin was selected for subsequent experiments. Compared with H 2O 2 groups, the cell proliferation activity in 40 and 60 μmol/L isornetin groups was significantly increased [(72.21±5.11)%, (76.08±4.91)%, P<0.05], SOD activity increased (19.81±0.38, 20.52±0.52, 15.45±3.13, P<0.05) and MDA content decreased (35.94±0.31, 22.04±0.26, 19.26±1.36, P<0.05). Conclusions:The flavonoid isorhamnetin has a protective effect on oxidative stress injury induced by H 2O 2 in HaCaT cells, suggesting that isorhamnetin may be a potential drug component in the treatment of vitiligo.

Objective:To investigate the present situation of unintended pregnancy within two years postpartum and its influencing factors in China.Methods:Participants who delivered a live birth at 60 hospitals in 15 provinces in the eastern, central and western regions of China during July 2015 to June 2016 were interviewed by using structured questionnaire. Information on occurrence of unintended pregnancy within 2 years after delivery, postpartum contraceptive use, sexual resumption, breastfeeding, and women′s socio-demographic characteristics, and so on, were collected. Life-table analysis, cluster log-rank tests and a 2-level Cox regression model were used for data analysis.Results:A total of 18 045 postpartum women were investigated. The cumulative 1- and 2-year unintended pregnancy rates after delivery were 5.3% (95% CI: 4.5%－6.1%) and 13.1% (95% CI: 11.3%－14.8%), respectively. Cox regression model analysis showed that the risk of unintended pregnancy within 2 years postpartum were increased in younger women, ethnic minorities, women with abortion history, and those who had a vaginal delivery with short lactation time and late postpartum contraceptive initiation (all P<0.01). The risk of postpartum unintended pregnancy was not associated with geographic regions and hospitals where women gave a birth (all P>0.05). Conclusions:In China, the risk of unintended pregnancy within 2 years after delivery is relatively high. Service institutions and service providers should improve the quality of postpartum family planning services, promote the use of high effect contraceptive methods, and educate women to use a method at the time of their sexual resumption or even before.