Objective: To explore the changes of mechanosensitive channels Piezos (Piezo 1 and Piezo 2) in neurogenic bladder (NB) of young rats and their effects，so as to provide reference for clinical search of new therapeutic targets. Methods: A total of 30 female young SD rats were divided into 5 groups based on random number table method：sham operation group (sham)，2-week nerve transection group (NB-2W)，6-week nerve transection group (NB-6W)，2-week nerve transection + Piezos inhibitor group (NB-P-2W) and 6-week nerve transection + Piezos inhibitor group (NB-P-6W)，with 6 rats in each group.The NB models were constructed by transecting the L6 and S1 spinal nerves of young rats.The NB-2W and NB-6W groups were not intervened after modeling，while the NB-P-2W and NB-P-6W groups were intraperitoneally injected with Piezos inhibitor GsMTx4 (10 mg/kg) every 2 days after modeling.Bladder cystometry and ultrasound were performed after 2 and 6 weeks of transection.The expressions of Piezos and fibrosis-related indexes (Collagen Ⅰ and α-smooth muscle actin) were detected in bladder tissues. Results: The results of bladder cystometry showed that the basal bladder pressure in NB-2W group was significantly increased，while it was slightly decreased but was still higher in NB-6W group than in the sham group (P<0.05).Basal bladder pressure was lower in NB-P-2W group than in NB-2W group，but was higher than that in the sham group; basal bladder pressure was lower in NB-P-6W group than in NB-6W group，but higher than that in the sham group (P<0.05).Compared with the sham group，the NB-2W and NB-6W groups had firstly increased and then decreased maximum cystometric capacity (MCC) (P<0.05).Compared with NB-2W group，NB-P-2W group had lower bladder leakage point pressure (BLPP)，but higher MCC and bladder compliance (BC) (P<0.05).Compared with NB-6W group，NB-P-6W group had significantly lower BLPP but higher MCC and BC (P<0.05).HE and MASSON staining and ultrasound results showed that，with the extension of nerve transection time，bladder fibrosis gradually worsened，the bladder wall became rough and thickened，calculi were visible inside，and hydronephrosis gradually appeared; the degree of fibrosis in NB-P-2W and NB-P-6W groups was less than that in NB-2W and NB-6W groups，and no hydronephrosis was observed in the upper urinary tract.In addition，Western blotting and immunohistochemical results showed that NB-2W and NB-6W groups had significantly higher relative expression levels of Piezos，Collagen Ⅰ and α-SMA than the sham group (P<0.01)，while NB-P-2W and NB-P-6W groups had lower relative expression levels of Piezos,Collagen Ⅰ and α-SMA than NB-2W and NB-6W groups (P<0.01). Conclusion: The increased expressions of mechanosensitive channels Piezos in NB young rats may be involved in the progression of bladder fibrosis，but its mechanism needs further study.

Cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a non-atherosclerotic, non-amyloidogenic inherited cerebral small vessel disease caused by mutations in the NOTCH3 gene, typically manifesting in middle age. The diagnosis can be confirmed through the detection of granular osmiophilic material in skin biopsies and NOTCH3 gene mutations identified by genetic testing. However, the pathogenesis of this disease remains unclear, and treatment options are limited. A major contributing factor is that currently available preclinical animal models fail to adequately recapitulate the characteristic pathological features and clinical phenotypes of the disease. Therefore, a comprehensive review of recent advances in CADASIL animal models was conducted, and the advantages and limitations of existing models were systematically analyzed, aiming to offer guidance for selecting appropriate animal models in fundamental research and to propose future directions for developing experimental models.

Cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a non-atherosclerotic, non-amyloidogenic inherited cerebral small vessel disease caused by mutations in the NOTCH3 gene, typically manifesting in middle age. The diagnosis can be confirmed through the detection of granular osmiophilic material in skin biopsies and NOTCH3 gene mutations identified by genetic testing. However, the pathogenesis of this disease remains unclear, and treatment options are limited. A major contributing factor is that currently available preclinical animal models fail to adequately recapitulate the characteristic pathological features and clinical phenotypes of the disease. Therefore, a comprehensive review of recent advances in CADASIL animal models was conducted, and the advantages and limitations of existing models were systematically analyzed, aiming to offer guidance for selecting appropriate animal models in fundamental research and to propose future directions for developing experimental models.

In the course of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infec-tion,to verify whether ursodeoxycholic acid(UDCA)can reduce angiotensin-converting enzyme 2(ACE2)receptor in BALB/c mice and reduce the risk of infection.UDCA was administered by in-tragastric administration to BALB/c mice for 7 d.During the treatment,the turbinate bones and lungs of mice were taken every day,and the changes of ACE2 content in the turbinate bones and lungs of mice were detected by ELISA.In addition,after 1,4 and 7 d of intragastric prophylaxis,BALB/c mice were infected with SARS-CoV-2 C57MA14 mouse adapted strain and SARS-CoV-2 Omicron DY1.1,respectively,after nasal inoculation,and viral load was detected on the turnings and lungs of mice 3 d after challenge to evaluate the preventive effect.In addition,UDCA was used to treat BALB/c mice infected with SARS-CoV-2 C57MA14 mouse adapted strain after nasal drops by gavage for 3 d,and the viral load of the mouse turbinate and lung was detected to evaluate the therapeutic effect.UDCA can decrease ACE2 content in turbinate and lung of BALB/c mice.How-ever,after 1,4 and 7 d of UDCA intragastric administration,there was no statistical difference in viral load in turbinate and lung of BALB/c mice between the prevention group and the virus con-trol group.There was no significant difference in the viral load of the turbinate and lung between the UDCA treatment group and the viral control group.UDCA could reduce the ACE2 content in the turnings and lungs of aged BALB/c mice,but the daily dose and duration of UDCA treatment had no significant effect on the mice infected with SARS-CoV-2 C57MA14 mouse adapted strain and SARS-CoV-2 Omicron DY1.1.

Objective:To investigate the changes in the morphology, structure and function of the bladders and their effects on the upper urinary tract dilatation(UUTD) after lumbosacral nerve transecting in rats.Methods:A total of 45 female SD rats were included, randomly divided into 3 groups with 15 rats in each group. Two groups were performed bilateral lumbar 6(L6) and cauda equina nerve shearing to establish neurogenic bladder(NB) model, which were nerve transected for 4 weeks(NB-4W) group and nerve transected for 12 weeks(NB-12W) group. Another group was performed bilateral L6 nerves and cauda equine exposing but not transecting, which was sham-operation (Sham) group. Cystometry and renal ultrasound examination were performed and rats in each group were killed to collect the kidney and bladder tissues in NB-4W group at 4 weeks, in Sham group and NB-12W group at 12 weeks after operation. HE, Masson staining, immunohistochemical staining and western blot were used to detect histological changes, expression of transforming growth factor-β1(TGF-β1) and α-smooth muscle actin(α-SMA).Results:All rats in NB-4W and NB-12W group showed acontractile detrusor. In the NB-4W and NB-12W group, the maximum cystometric capacity [(5.84±0.33) ml and (3.13±0.35) ml], the detrusor leak point pressure [(25.41±0.86) cm H 2O and (27.36±2.04) cm H 2O] (1 cm H 2O = 0.098 kPa), were significantly higher than those in the Sham group [(0.98±0.14) ml, (7.13±0.90) cm H 2O, both P<0.05]. Compliance in NB-4W group [(0.28±0.21) ml/cm H 2O] and NB-12W group [(0.17±0.12) ml/cm H 2O] were significantly lower than that of the Sham group [(0.34±0.26) ml/cm H 2O], and the compliance of NB-12W group was lower than that of NB-4W group significantly (all P<0.05). HE staining of the bladder showed that the inflammatory cell infiltration was obvious in the NB-4W and NB-12W group. Bladder collagen volume fractions in NB-4W group [(30.5±1.5) %] and NB-12W group [(45.2±3.8) %] were both higher than that of Sham group [(20.7±2.2) %, both P<0.05]. The expression of TGF-β1 and α-SMA in the bladder tissue of NB-4W group were higher than those of sham group, and that of NB-12W group were higher than NB-4W group. In NB-4W group and NB-12W group, 3 (20.0 %) and 7 (46.7 %) rats were found hydronephrosis, respectively. Additionally, HE staining showed that the degree of renal tubule injury and the number of inflammatory cell infiltration in the NB-4W and NB-12W group were higher than those in the Sham group. Masson staining showed that the volume fraction of collagen in kidneys of NB-4W and NB-12W group were (13.1±1.4) % and (21.6±1.9) %, respectively, which were significantly higher than that in sham operation group [(4.6±0.7) %, both P<0.05]. Conclusions:Bilateral L6 + cauda equina nerve transecting can induce NB with hydronephrosis in parts of rats. The degree of bladder fibrosis gradually increased with the time of nerve transection, and the incidence and severity of UUTD also increased with the time of nerve transection.

Megalencephalic leukoencephalopathy with subcortical cysts (MLC, OMIN: 604004) caused by mutations in the MLC1 gene, is an rare autosomal recessive disorder. More patients are with infancy and young children onset, whereas adult cases are rare. Only 2 patients from 1 family have been reported in domestic adult cases. Now a 58-year-old female MLC patient is reported. The clinical manifestations of the patient included large head circumference, slow responses, walking difficulties, seizures and paroxysmal loss of consciousness. The result of whole exome sequencing revealed a homozygous insertion mutation c.920_943dup in the MLC1 gene. The mutation in this patient has not been reported in the Human Gene Mutation Database.

Multiple sclerosis (MS) is an idiopathic inflammatory demyelinating diseases of the central nervous system, the underlying cause of which has not been cleared. Previous studies have shown that the pathogenesis of MS is related to the destruction of blood brain barrier, furthermore the drugs used to treat MS have a certain protective effect on the function of blood brain barrier. Therefore, this review combines the research progress at home and abroad to clarify the relationship between the blood brain barrier and MS in pathogenesis and treatment, proposing possible orientation of development.

Objective To observe the effects of neurotrophin 3 (NT3)-chitosan on motor function, and proliferation and differentiation of the neural stem cells (NSCs) in the injury area and subventricular zone (SVZ) in rats with motor cortex injury. Methods Sixty-five Wistar rats were divided into control group (n=7), injury group (n=29) and NT3-chitosan group (n=29). The motor cortex was aspirated and re-moved as cerebral injury model. NT3-chitosan was immediately implanted into the injured area after operation, and the control group re-ceived no intervention. Pellet reaching test was performed to detect the recovery of the forelimb function, HE staining was used to observe the lesion cavity size, and immunofluorescence staining was used to observe the proliferation and differentiation of NSCs 3 days, 7 days, 14 days, 1 month, 2 months and 3 months after operation. Results The grasp success rate was higher (F>6.00, P≤0.05), and the lesion cavity size was significantly smaller (F>629.5, P<0.001) in the NT3-chitosan group than in the injury group. In the NSCs differentiation experi-ment, the number of BrdU cells at all the time points was significantly higher in the NT3-chitosan group than in the injury group (F>171.43, P<0.001). In the NSCs proliferation experiment, the number of BrdU positive cells was still significantly higher in the NT3-chitosan group than in the control group and in the injury group (F>155.06, P<0.001), the number of Dcx positive cells was significantly higher in the NT3-chitosan group than in the injury group (F=62.367, P<0.001), and the number of BrdU/Dcx positive cells was significantly higher in the NT3-chitosan group than in the control group (F=33.527, P<0.001). Conclusion NT3-chitosan could activate NSCs in the SVZ, and pro-mote endogenous neurogenesis and forelimb function recovery in rats after motor cortex injury.

AIM:To study the effects of extracellular potassium on the protein expression of wild-type HERG and its mutant L539fs/47.METHODS:Wild-type HERG (WT) or its mutant HERG-L539fs/47 (MT) were transfected into HEK293 cells for 36 h.The cells were incubated in different media containing 0.8, 4.3 or 10 mmol/L potassium.Af-ter 6 h of incubation, the protein expression of HERG was detected by flow cytometry.After 12 h of incubation, the locali-zation and quantity of the proteins were detected by laser confocal imaging and Western blot.RESULTS: Different from the retention of mutant protein in cytoplasm, wild-type HERG protein was mainly distributed in the cell membrane.The 2 proteins both increased with the changes of extracellular potassium.Flow cytometry showed that the fluorescence in the 2 groups both increased with the changes of extracellular potassium ( P<0.01 ) .The fluorescence in WT group was signifi-cantly higher than that in MT group (P<0.01).Western blot showed that mutant HERG protein included only one 60 kD band, different from the 135 kD and 155 kD bands in wild-type HERG, which were affected by the changes of extracellular potassium (P<0.05).CONCLUSION:The retention of HERG mutant L539fs/47 protein in the cytoplasm is more than wild-type HERG.Chronic high extracellular potassium keeps the stability of wild-type and mutant HERG proteins on the cell membrane.Chronic low potassium reduces the expression of HERG channel proteins in a time-dependent manner.

Objective To analyze the clinical effect of the bone grafting pedicled with femoral quadratus for the osteonecrosis of femoral head.Methods The clinical operation data were analyzed from 355 patients with osteonecrosis of femoral head underwent by the surgery of the bone grafting pedicled with femoral quadrates in the First Affiliated Hospital of Zhengzhou University from August,2006 to June,2013.According to association research circulation osseous (ARCO) classifying,41,126,115,62,6 and 5 patients were in stage Ⅰ-C,Ⅱ-A,Ⅱ-B,Ⅱ-C,Ⅲ-A,and Ⅲ-B.The results were evaluated by the hundred forked method.Results All the patients obtained the outpatient followed-up after operation,the follow-up time was 1-8 (4.3 ± 1.8) years.The pain of the hip,the function of the joint,the motion of hip joint,and the X ray evaluation became better than preoperations.There were statistically significant differences in the healing time among all the age patients (P ＜ 0.05),along with the age growth,the healing time gradually extended.Compared with that before operation,the scores of all the stage patients after operation were higher (P ＜ 0.05).The rate of excellent and good results were 98.5％,98.0％,92.3％,89.7％,83.5％,81.3％ in stage Ⅰ-C,Ⅱ-A,Ⅱ-B,Ⅱ-C,Ⅲ-A,and Ⅲ-B,the excellent and good rate was 93.8％ of all the 355 patients.Conclusion Bone grafting pedicled with femoral quadratus is effective surgical method for the osteonecrosis of femoral head.