When partial nephrectomy is performed by posterior abdominal approach, the surgical field is poorly exposed, resulting in increased surgical difficulty and risk of injury.In this study, 28 patients with T 1a stage kidney tumors underwent retroperitoneal laparoscopic partial nephrectomy. Intraoperatively, exposure of the surgical field was achieved using the percutaneous puncture of the renal fascia suspension technique. There were no dissatisfactory exposures due to peritoneal damage during the surgery, no additional tubes were inserted, and no conversions to open surgery were needed. The operation time was (76.5±20.3) minutes, blood loss was (92.1±18.7) ml, renal artery clamping time was (19.5±4.3) minutes. Postoperatively, there were no complications such as bleeding, infection, or hematuria.

The assessment of adverse drug events is an important basis for clinical safety evaluation and post-marketing risk control of drugs,and its causality assessment is gaining increasing attention.The existing methods for assessing the causal relationship between drugs and the occurrence of adverse reactions can be broadly classified into three categories:global introspective methods,standardized methods,and probabilistic methods.At present,there is no systematic introduction of the operational details of the various methods in the domestic literature.This paper compares representative causality assessment methods in terms of definition and concept,methodological steps,industry evaluation and advantages and disadvantages,clarifies the basic process of determining the causality of adverse drug reactions,and discusses how to further improve the adverse drug reaction monitoring and evaluation system,with a view to providing a reference for drug development and pharmacovigilance work in China.

Objective To investigate the effects and the mechanisms of sulforaphane(SFN)on endothelin-1(ET-1)-induced proliferation in rat vascular smooth muscle cell(VSMC).Methods Rat VSMC were cultured by using the modified tissue explant technique in vitro and were randomly divided into control group(normal culture),model group(treated with 0.1 μmol·L-1 ET-1)and experimental-L,-M,-H groups(treated with 5,10,20 SFN+0.1 μmol·L-1 ET-1).The survival rate of VSMC were measured by methyl thiazolyl tetrazolium.The proliferation and apoptosis related genes in mRNA level in VSMC were respectively examined with real-time quantitative polymerase chain reaction;the relative fluorescence intensity of reactive oxygen species(ROS)in VSMC were measured by fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate;the protein expression of B-cell lymphoma-2(Bel-2),cysteinyl aspartate-specific proteinase-3(caspase-3),p38 mitogen-activated protein kinases(p38MAPK),phosphorylated p38 mitogen-activated protein kinases(p-p38MAPK),and p53 in VSMC were investigated by Western blot.Results The proliferation activities of the control group,model group,experimental-L,-M,-H groups were 0.27±0.02,0.50±0.04,0.40±0.03,0.38±0.03 and 0.34±0.03;the expression levels of Bcl-2 mRNA were 1.57±0.28,1.00±0.00,0.87±0.05,0.75±0.08 and 0.41±0.18;the expression levels of caspase-3 mRNA were 0.84±0.10,1.00±0.00,1.33±0.10,1.61±0.15 and 1.83±0.16;the relative fluorescence intensities of ROS in VSMC were 6.05±1.50,31.45±3.12,30.03±1.85,18.39±5.62 and 17.18±1.97;the relative expression levels of Bel-2 protein were 0.65±0.02,0.60±0.02,0.49±0.02,0.48±0.03 and 0.49±0.01;the relative expression levels of caspase-3 protein were 0.03±0.00,0.25±0.01,0.42±0.01,0.46±0.02 and 0.64±0.03;the protein expression ratios of p-p38MAPK/p38MAPK were 0.97±0.05,1.44±0.04,1.62±0.10,2.18±0.05 and 2.70±0.05;the relative expression levels of p53 protein were 0.20±0.01,0.30±0.01,0.34±0.02,0.37±0.01 and 0.42±0.01 respectively.There were statistically significant differences in the above indicators between model group and control group,between model group and experimental-M,-H groups(P＜0.05,P＜0.01).Conclusion These demonstrate that SFN may inhibit proliferation and prompt apoptosis in ET-1-stimulated VSMC by anti-oxidant and activating p38MAPK and p53-depended apoptosis signal pathway.

The National Medical Products Administration have become an official applicant of the Pharmaceutical Inspection Co-operation Scheme(PIC/S)in 2023,and the good manufacturing practice appendix and inspection memorandum are key documents of PIC/S.Based on the analysis of the history,framework and main contents of the appendix and inspection memorandum of PIC/S investigational medicinal products,this paper discusses the key contents of the appendix and inspection memorandum of PIC/S clinical trial drugs,and provides reference for the inspection work.

Objective:To investigate the effect of selenium on cyclophosphamide(CTX)-induced spermatogenic impairment(SI)in mice and its underlying mechanism.Methods:We equally randomized 36 male KM mice into 3 SI model and 3 control groups,the first 3 treated by intraperitoneal injection of CTX at 100 mg/kg(the SI model control group),CTX plus SI model control group,selenium deficient model group(-Se SI),selenium supplemented model group(+Se SI),while latter 3 by intraperitoneal injection of normal saline(the normal control),selenium deficiency control group(-Se control),selenium addition control group(+Se control),respectively,all once a week for 6 successive weeks.Then we observed the histopathological changes in the testes of all the mice by HE staining,obtained the sperm count in the epididymides,determined the expressions of glutathione peroxidase 4(GPx4)and SLC7A11 proteins by Western blot and ferroptosis-related genes by RT-qPCR,and examined the changes in the expres-sions of ferroptosis-related proteins and genes in the GC2-spd cells treated with ferroptosis inhibitors and inducers in combination with different concentrations of inorganic sodium selenite(SeS)and organic selenomethionine(SeM).Results:Compared with the nor-mal controls,the SI model mice showed significantly decreased testicular and prostatic organ coefficients,reduced spermatogenic lay-ers,increased voids,decreased serum ferritin concentration(P<0.05),and elevated transferrin concentration(P<0.05).The or-gan coefficients were significantly higher in the+Se SI and+Se control than in the-Se SI and-Se control groups(P<0.05,P<0.01),with evident pathological improvement of the testis tissue in the+Se controls.The expressions of the GPx4 and solute carrier family 7 members 11(SLC7A11)genes in the testis were dramatically down-regulated in the SI model controls(P<0.01),but up-reg-ulated in the+Se SI and+Se control compared with those in the-Se SI and-Se control group(P<0.01 and P<0.05),but there were no statistically significant differences between their protein expressions.The results of in vitro GC2 spd cell experiments indicated that the GPx4 gene and GPx4 protein levels in the-Se group were significantly lower than those in the normal control group(P<0.05),while the SLC7A11 gene level decreased(P<0.01).Different doses of SeS and SeM significantly increased the GPx4 protein expression compared to the average Se group.Low doses of SeM promoted a significant increase in GPx4 gene levels,while high doses of SeS increased the expression levels of SLC7A11 gene and SLC7A11 protein(P<0.05,P<0.01).The Se group showed a signifi-cant decrease in the levels of acsl4 and ptgs2 genes compared to the normal control group.SeM promoted the expression of acsl4,while SeS promoted the expression of ptgs2 and fth1(P<0.01,P<0.05).The intervention results of GC2 spd showed that the Erastin group had a decrease in ptgs2 compared to the normal control group,while the SeS+Erastin and SeM+Erastin groups had an increase in ptgs2 gene expression compared to the Erastin group.However,the ptgs2 expression of Fer-1 was lower than that of the normal con-trol group,and the ptgs2 gene level of SeS+Fer-1 and SeM+Fer-1 groups was lower than that of Fer-1 group(P<0.05);The gene quantity of GPx4 in the SeM+Erastin and SeM+Fer-1 groups increased compared to the Erastin and Fer-1 groups(P<0.01,P<0.05);SeM+Erastin and SeS+Erastin showed a decrease in SLC7A11 compared to the Erastin group,as well as SeM+Fer-1 and SeS+Fer-1 groups compared to the Fer-1 group,accompanied by an increase in acsl4 and fth1(P<0.01).Conclusion:Selenium deficiency causes the reduction of the SLC7A11 and GPx4 gene levels,disorder of ferroptosis-related genes and down-regulation of the GPx4 protein expression in the mouse testis and spermatocytes.Selenium can promote the expression of GPx4,up-regulate the level of SLC7A11,and improve spermatogenesis in the testis of the mouse with SI.There are differences between organic SeM and inorganic SeS in regulating the ferroptosis pathway-related genes.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

Objective To observe the effects of melatonin on autophagy in cortical neurons of neonatal rats with hypoxic-ischemic brain damage(HIBD)and to explore its mechanisms via the PI3K/AKT signaling pathway,aiming to provide a basis for the clinical application of melatonin.Methods Seven-day-old Sprague-Dawley neonatal rats were randomly divided into a sham operation group,an HIBD group,and a melatonin group(n=9 each).The neonatal rat HIBD model was established using the classic Rice-Vannucci method.Neuronal morphology in the neonatal rat cerebral cortex was observed with hematoxylin-eosin staining and Nissl staining.Autophagy-related protein levels of microtubule-associated protein 1 light chain 3(LC3)and Beclin-1 were detected by immunofluorescence staining and Western blot analysis.Phosphorylated phosphoinositide 3-kinase(p-PI3K)and phosphorylated protein kinase B(p-AKT)protein expression levels were measured by immunohistochemistry and Western blot.The correlation between autophagy and the PI3K pathway in the melatonin group and the HIBD group was analyzed using Pearson correlation analysis.Results Twenty-four hours post-modeling,neurons in the sham operation group displayed normal size and orderly arrangement.In contrast,neurons in the HIBD group showed swelling and disorderly arrangement,while those in the melatonin group had relatively normal morphology and more orderly arrangement.Nissl bodies were normal in the sham operation group but distorted in the HIBD group;however,they remained relatively intact in the melatonin group.The average fluorescence intensity of LC3 and Beclin-1 was higher in the HIBD group compared to the sham operation group,but was reduced in the melatonin group compared to the HIBD group(P<0.05).The number of p-PI3K+and p-AKT+cells decreased in the HIBD group compared to the sham operation group but increased in the melatonin group compared to the HIBD group(P<0.05).LC3 and Beclin-1 protein expression levels were higher,and p-PI3K and p-AKT levels were lower in the HIBD group compared to the sham operation group(P<0.05);however,in the melatonin group,LC3 and Beclin-1 levels decreased,and p-PI3K and p-AKT increased compared to the HIBD group(P<0.05).The correlation analysis results showed that the difference of the mean fluorescence intensity of LC3 and Beclin-1 protein in the injured cerebral cortex between the melatonin and HIBD groups was negatively correlated with the difference of the number of p-PI3K+and p-AKT+cells between the two groups(P<0.05).Conclusions Melatonin can inhibit excessive autophagy in cortical neurons of neonatal rats with HIBD,thereby alleviating HIBD.This mechanism is associated with the PI3K/AKT pathway.

Objective To investigate the clinical features and prognosis of childhood acute lymphoblastic leukemia (ALL) with CREBBP gene mutation. Methods A retrospective analysis was performed for the clinical data of 14 ALL children with CREBBP gene mutation who were admitted to Children's Hospital of the First Affiliated Hospital of Zhengzhou University from January 2016 to December 2023. Results The ALL patients with CREBBP gene mutation accounted for 1.5％ (14/963) among all children diagnosed with ALL during the same period of time,among whom there were 4 boys (29％) and 10 girls (71％),with a median age of 4 years and 3.5 months. All children had an immunological type of B-cell ALL and concurrent mutations in other genes including NRAS,KRAS,ETV6,FLT3,PAX5,SH2B3,CDKN2A,and CDKN2B,and 4 children had karyotype abnormality. All 14 children received induction therapy with the VDLP regimen,with a complete remission (CR) rate of 79％ (11/14) after the first course of treatment. Three children experienced bone marrow recurrence alone,with a recurrence rate of 21％ (3/14),among whom 1 child achieved CR after blinatumomab therapy and 2 received bridging hematopoietic stem cell transplantation after chemotherapy for recurrence. Among the 14 children,1 died due to treatment discontinuation and 13 achieved disease-free survival. The 5-year overall survival rate was 92％±7％,and the event-free survival rate was 73％±13％. Conclusions ALL with CREBBP gene mutation is more common in girls and has a low induction remission rate and a high recurrence rate,and it is often accompanied by other types of gene mutations and abnormal karyotypes. Most children with recurrence can achieve long-term survival after immunotherapy or hematopoietic stem cell transplantation.

Objective To observe the clinical efficacy of modified Dahuang Fuzi Decoction following the therapy of unblocking bowels for the treatment of acute pancreatitis of pathogenic-cold accumulation and retention type.Methods A total of 78 patients with acute pancreatitis of pathogenic-cold accumulation and retention type were randomly divided into a study group and a control group,39 cases in each group.The control group was treated with conventional western medicine,and the study group was treated with oral use or nasogastric feeding of Dahuang Fuzi Decoction on the basis of treatment for the control group.The course of treatment covered seven days.The two groups were observed in the changes of serum pancreatic enzyme indicators of serum amylase and lipase,inflammatory factors of C-reactive protein(CRP),procalcitonin(PCT),interleukin-6(IL-6)and interleukin-8(IL-8),acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)score,and modified CT severity index(MCTSI)score before and after treatment.The time for abdominal pain relief,time for abdominal distension relief,and time for anal defecation were compared between the two groups.After treatment,the clinical efficacy and safety of the two groups were evaluated.Results(1)During the trial,there was one patient falling off separately from the study group and the control group,and 38 patients in each group were eventually included in the efficacy statistics.(2)After seven days of treatment,the total effective rate of the study group was 86.84％(33/38),and that of the control group was 78.95％(30/38).The intergroup comparison(tested by rank sum test)showed that the efficacy of the study group was significantly superior to that of the control group(P＜0.05).(3)After treatment,the serum levels of pancreatic enzyme indicators of amylase and lipase in the two groups were decreased compared with those before treatment(P＜0.05),and the decrease of serum amylase and lipase levels in the study group was significantly superior to that in the control group(P＜0.05).(4)After treatment,the serum levels of inflammatory factors of CRP,PCT,IL-6 and IL-8 in the two groups were decreased compared with those before treatment(P＜0.05),and the decrease of serum CRP,PCT,IL-6 and IL-8 levels in the study group was significantly superior to that in the control group(P＜0.05).(5)After treatment,the APACHE Ⅱ and MCTSI scores of the two groups were decreased compared with those before treatment(P＜0.05),and the decrease of APACHE Ⅱ and MCTSI scores in the study group was significantly superior to that in the control group(P＜0.05).(6)For the time of symptom relief,the time for abdominal pain relief,time for abdominal distension relief,and time for anal defecation in the study group were significantly shortened compared with those in the control group,and the differences were statistically significant(P＜0.05).(7)No obvious adverse reactions occurred in the two groups during the treatment,indicating high safety.Conclusion On the basis of conventional treatment,the use of modified Dahuang Fuzi Decoction following the therapy of unblocking bowels exerts certain efficacy for the treatment of patients with acute pancreatitis of pathogenic-cold accumulation and retention type.The therapy is effective on relieving the symptoms and signs of patients,and decreasing the level of serum pancreatic enzyme indicators and inflammatory factors.

Objective To investigate the dynamic expression with the time change of N6-methyladenosine(m6A)methylation-related factors in the repair process of skeletal muscle injury and its mechanism in the inflammatory response of macrophage in the injure process.Methods In vivo mice models of BaCl2 injury in the gastrocnemius were established.Four mice per group in the control group and injury group.Gastrocnemius tissues were harvested at day 1,3,5,7,and 9 after injury for experiments.Primary gastrocnemius muscle tissue cells,muscle satellite cells,muscle cells,and cell line C2C12 cells were treated with dexamethasone(DEX,50 μmol/L)to mimic injury.Lipopolysaccharide(LPS,100 μg/L)induced RAW264.7 cell lines to mimic the inflammatory response after skeletal muscle injury,and STM2457(30 μmol/L)was added to inhibit the effect of methyltransferase 3(Mettl3)before LPS treatment.The expression of m6A methylation-related factors(Writers,Erasers,Readers)and inflammation factors were detected by Real-time PCR and Western blotting.Results The muscle fibers were dissolved and then gradually repaired with the extension of injury time,the number of monocytes/macrophages increased first and then decreased,and the Pax7 mRNA level increased first and then decreased with the change of injury time.Compared with the control group,the mRNA and protein levels of m6A methylation-related factors in gastrocnemius did not change significantly on the injury-1 day.However,they were significantly increased on the injury-3 days compared with the control group(P＜0.05),and then obviously decreased on the injury-5 days group compared with the injury-3 days group(P＜0.05).Compared with the control group,they were no significant differences on the injury-7 days group and-9 days group.In vitro DEX decreased the mRNA levels of m6A methyltransferase factors in primary muscle satellite cells and C2C12 cells and increased the mRNA expression level of methylation-recognition enzyme factors(P＜0.05).The mRNA levels of m6A methylation-related factors increased significantly in skeletal muscle tissue cells and myocytes after DEX treatment(P＜0.05).After LPS treatment,the mRNA and protein expression levels of m6A methylation-related factors and the mRNA expression levels of inflammatory factors interleukin(IL)-6 and IL-1β in macrophages increased significantly(P＜0.05),while the levels of IL-6 and IL-1β mRNA in macrophages decreased significantly when the Mettl3 was inhibited(P＜0.05).Conclusion m6A methylation-related factors primarily is activated in the damaged muscle cells and inflammation response of macrophages.Inhibition of m6A methyltransferase can reduce the inflammatory response of macrophages.