BACKGROUND:Inflammation is one of the important factors that induce cerebral ischemia-reperfusion injury.Studies have shown that electroacupuncture can effectively reduce inflammation after ischemic stroke and improve the symptoms of neurological deficits,but the mechanism is not clear. OBJECTIVE:To observe the effect of electroacupuncture on Toll-like receptor 4/nuclear factor-κB in rats with cerebral ischemia-reperfusion injury. METHODS:Forty-eight male Sprague-Dawley rats were randomly divided into sham operation group,model group and electroacupuncture group,with 16 rats in each group.The rat model of cerebral ischemia-reperfusion injury was prepared by middle cerebral artery occlusion.At 24 hours after modeling,the rats in the electroacupuncture group were treated with electroacupuncture,once a day,20 minutes each time,for a total of 5 days.The sham operation group and the model group did not do any intervention.After 5 days of intervention,Longa method was used to evaluate the degree of neurological injury in rats.Triphenyl tetrazolium chloride staining and hematoxylin-eosin staining were used to measure the volume of cerebral infarction and the pathological changes of brain tissue in rats.Serum interleukin-6,interleukin-18 and tumor necrosis factor-α were detected by ELISA.Expressions of Toll-like receptor 4 and nuclear factor-κB in the cerebral cortex at mRNA and protein levels were detected by fluorescence quantitative PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the sham operation group,the neurological function scores,serum interleukin-6,interleukin-18,and tumor necrosis factor-α levels,Toll-like receptor 4 and nuclear factor-κB mRNA and protein expression levels were significantly higher in the model group(P＜0.01).Compared with the model group,electroacupuncture significantly reduced the neurological function scores,serum interleukin-6,interleukin-18,and tumor necrosis factor-α levels,Toll-like receptor 4 and nuclear factor-κB mRNA and protein expression levels(P＜0.05,P＜0.01).Compared with the sham operation group,the volume of cerebral infarction in the model group increased significantly(P＜0.01).Compared with the model group,the volume of cerebral infarction in the electroacupuncture group decreased(P＜0.05).In the model group,the arrangement of neurons was disordered,some nerve cells disappeared,nuclei presented with pyknosis and incomplete structure.After electroacupuncture intervention,the degree of neuronal degeneration and neuronal loss in the cerebral cortex of rats were reduced compared with those in the model group.To conclude,electroacupuncture can significantly improve the neurobehavior of rats with cerebral ischemia-reperfusion injury,reduce brain tissue injury,and effectively reduce the level of serum inflammatory factors.The mechanism may be related to the inhibition of Toll-like receptor 4/nuclear factor-κB signaling pathway.

Objective: To identify and analyze the variants of the KCNJ1 gene in five Chinese patients with Bartter syndrome type 2 (BS2), and to describe their clinical features as well as treatment results. Methods: Data and blood samples of five BS2 patients and their relatives confirmed by Qingdao Municipal Hospital from June 2012 to January 2019 were collected. Whole-exome-sequencing (WES) based on the second generation high throughput sequencing was performed to detect variants. The 2015 American College of Medical Genetics and Genomics Standards and Guidelines were applied to analyze the pathogenicity of the variants. The clinical features and laboratory results were retrospectively studied. The response to treatment and follow-up data were reviewed. Results: Ten variants including six novel ones of KCNJ1 gene were identified through WES and verified by Sanger dideoxy sequencing. Missense variants accounted for the highest proportion. The common symptoms and signs of five BS2 patients from high to low incidence were polydipsia and polyuria (5/5), one of them (1/5) presented with diabetes insipidus; maternal polyhydramnios and premature delivery (4/5); growth retardation (3/5). Initially, two patients presented with hypochloremic metabolic alkalosis and hypokalemia, whereas the acid-base disturbance was absent in the others. One patient experienced hyperkalemia. In terms of calcium-phosphorus metabolism, one patient had evident parathyroid hormone (PTH) resistance (hypocalcemia, hyperphosphatemia and markedly elevated serum intact PTH levels), three presented with PTH overacting (hypercalcemia, hypophosphatemia and mild elevated serum intact PTH levels), and one showed normal blood calcium and phosphorus concentrations with high-normal serum intact PTH levels. All patients had nephrocalcinosis or hypercalciuria, and one of them complicated with nephrolithiasis. Indomethacin helped to correct the growth retardation, halt polydipsia polyuria, decrease the elevated urinary calcium excretion, and normalize electrolyte disturbance as well as PTH parameters in some patients. Conclusions This investigation identifies ten variants of KCNJ1 gene, including six ones that have not been previously reported, which will enrich the human gene mutation database (HGMD). These patients in our study have atypical BS phenotype, so that careful differentiation from other parathyroid diseases will be required for clinicians.

Objective: To investigate the efficacy of enticavir and lamivudine in preventing rituximab-associated hepatitis B virus(HBV) reactivation in patients with B-cell non-Hodgkin lymphoma complicated with resolved hepatitis B during chemotherapy. Methods: This retrospective study included 216 B-cell non-Hodgkin lymphoma patients with complete data from January 2012 to January 2018 treated in 3 hospitals.Of 78 patients with resolved hepatitis B, they were divided into lamivudine prophylactic group(17 cases), entecavir prophylactic group(11 cases) and control group(50 cases). The changes of HBVM, HBV DNA and liver function before or after rituximab combination chemotherapy were analyzed.The incidence of HBV reactivation, liver function injury and chemotherapy delay were compared. Results: Compared to the other 71 patients, 7 cases experienced HBV reactivation in 78 patients with resolved hepatitis B. There were no statistically significant differences between the two groups in patient demographics, pathological pattern, chemotherapy regimen.Six patients in the control group developed HBV reactivation(12%) and 1 patient in lamivudine prophylactic group(5.88%), none had HBV reactivation in enticavir prophylactic group.There was statistically significant difference among three groups(Fisher P=0.016). A total of 7 cases experienced HBV reactivation in 78 patients with resolved hepatitis B respectively, 2 cases in the first chemotherapy period, 2 cases in the third chemotherapy period, 1 case in the fifth chemotherapy period, the last one occurred after the completion of chemotherapy.Four patients had HBsAg reverse seroconversion, occurred in the 1, 3, 5 cycles during and after the completion of chemotherapy.Twenty patients (40.0%) experienced liver function parameters abnormal in the control group, 4 cases(23.5%) in lamivudine prophylaxis group, 2 cases (18.2%) in enticavir prophylaxis group during chemotherapy. Conclusion Antiviral prophylaxic therapy can potentially prevent rituximab associated HBV reactivation in patients with resolved hepatitis B. Entecavir can more reduce the risk of rituximab associated HBV reactivation than lamivudine.

To study the protective effect of Xingnaojing Injection on early global brain ischemia-induced deep coma in rats.
 Methods: The deep coma model was induced by global brain ischemia by using four-vessel occlusion method in male SD rats. According to the body weight, the rats were randomly divided into 8 groups: a model control group, three different dose of Xingnaojing Injection (1.8, 3.6 and 5.4 mL.kg-1) groups, a Xingnaojing Injection (3.6 mL.kg-1) plus PI3K inhibitor group, a naloxone injection (0.04 mL.kg-1) group and a naloxone injection (0.04 mL.kg-1) plus Xingnaojing Injection (3.6 mL.kg-1) group (n=8 per group). In addition, eight animals served as the sham group were performed same operation with the model group excepting no blockage of the blood vessels. After the operation, three different doses of Xingnaojing Injection and/or naloxone injection were given intravenously once a day for three days. Ten μL PI3K inhibitor (LY294002, 10 mmol/L) was injected via anterior cerebral ventricle at once after global brain ischemia. The awakening time after the first drug treatment, the grasping power and the autonomous activity within 10 min after the last drug treatment were recorded. The levels of both dopamine (DA) and glutamate (Glu) in cerebrospinal fluid were detected by ELISA. The pathological changes were observed in brain tissue slices with HE staining and the protein levels of Akt/p-Akt and cAMP-response element binding protein (CREB)/p-CREB in hippocampus were detected by Western blotting.
 Results: Comparing with the model group, single administration of Xingnaojing Injection could significantly shorten the waking time (P<0.05) and continuous administration of Xingnaojing Injection for 3 d could increase grasping power, distance, frequency and duration of autonomous activities (P<0.05 or P<0.01) in the deep coma rat. Also, Xingnaojing Injection could inhibit these increases in neurotransmitters DA and Glu contents (P<0.05 or P<0.01), and improve pathological changes of hippocampal tissue. Xingnaojing Injection significantly induced protein phosphorylation of both Akt and CREB (P<0.05 or P<0.01); this effect was inhibited by PI3K inhibitor (P<0.05 or P<0.01). Moreover, the protective effects of naloxone on awakening time, grasping power, the autonomous activity and hippocampus damage in global brain ischemia-induced deep coma could be enhanced by joint use of Xingnaojing Injection (P<0.05 or P<0.01).
 Conclusion: Xingnaojing Injection could significantly improve deep coma induced by global brain ischemia in rat, which is related to inducing PI3K/Akt-dependent protein phosphorylation of CREB, and reducing hippocampal damage. The protective effect of Xingnaojing Injection is synergistically enhanced by naloxone.
Animals ; Brain ; Brain Ischemia ; Coma ; Drugs, Chinese Herbal ; Male ; Phosphatidylinositol 3-Kinases ; Rats ; Rats, Sprague-Dawley

Objective To analyze the mutations of SLC12A1 gene in nine Chinese families with Bartter syndrome type I (BS1),and analyze the relationship between genotype and phenotype.Methods The next generation sequencing was used to detect mutations in nine BS1 patients including eight with antenatal BS (aBS) and one with classical BS (cBS).Clinical characteristics and biochemical findings at the first admission as well as follow-up were reviewed.Results 15 different mutations of SLC12A1 gene were identified,including 11 novel ones.Among nine probands,seven were compound heterozygotes,two were homozygotes.All patients presented with polydipsia and polyuria,and eight with growth retardation.All patients had lower than-normal serum chloride concentration,metabolic alkalosis,and elevated basal renin activity and aldosterone,and seven had hypokalemia.Through treatment of indomethacin and/or potassium chloride,biochemical indicators could roughly restored normal.Conclusion These findings will enrich the human gene mutation database (HGMD) and provide valuable references to the genetic counseling and diagnosis for Chinese population.

Objective To analyze and identify the mutations in SGLT2 gene of nine Chinese families with FRG, and determine the renal threshold for glucose excretion (RTG), so as to explore the association of genotype and RTG. Methods All coding regions of SGLT2 gene, including intron exon boundaries, were analyzed using PCR followed by direct sequence analysis. Quantitative test for 24?hour urine glucose and RTG were measured among 9 probands (21 patients) and their family members from 9 pedigrees (total 25 subjects). The differences in renal glucose thresholds between patients with different genotypes (heterozygotes and compound heterozygotes; c.886(-10_-31) del heterozygotes and other heterozygotes) were compared. Results Twelve mutations were identified by SGLT2 gene analysis, including 10 novel ones that were not included in HGMD:c.331T>C, p.W111R;c.374T>C, p.M125T; c.394C>T, p.R132C; c.612G>C, p.Q204H; c.829C>T, p.P277S; c.880G>A, p.D294N;c.1129G>A, p.G377S; c.1194C>A, p.F398L; c.1540C>T, p.P514S; c.1573C>T, p.H525Y. In thisstudy, the mutation c.886(-10_-31)del that is specific to Chinese population accounted for about 28％of the total alleles (5/18). The RTG values of patients with compound heterozygous mutations were much lower than those with simple heterozygous mutations [(1.28 ±0.10) vs (5.14±0.77) mmol/L; P<0.001];and c.886(-10_-31)del heterozygotes had significant lower RTG values than others [(4.43 ± 0.37) vs (5.70 ± 0.51) mmol/L, P<0.001]. Conclusions Ten novel mutations which may be related to FRG are found in this study, and c.886(-10-31)del may be a hot?spot mutation in Chinese patients. Compound heterozygotes had much lower RTG values than simple heterozygotes.

Objective To analyze the clinical features,diagnosis and treatment of testicular torsion.Methods The clinical data of 95 patients with testicular torsion admitted from March 2001 to April 2017 were analyzed retrospectively.Results The age of patients ranged from 13 to 42 years (mean 19.6 years).Among 95 patients,91 cases had essential clinical symptoms of scrotal pain;1 case reported the right testicular enlargement without pain,and 3 cases presented with left inguinal mass.Color Doppler flow imaging (CDFI) was performed in 91 patients,reduction or loss of blood flow signals were detected in 88 cases,and normal blood flow signals were presented in 3 cases.The diagnosis of testicular torsion was confirmed by CDFI in 91 cases before surgery,and 4 cases were diagnosed by medical history and physical examination.The duration from onset to surgery was ＜ 5 h in 10 cases,5-10 h in 25 cases,and ＞ 10 h in 60 cases.All the patients received operation,92 cases were diagnosed as testicular torsion in scrotum and 3 cases as cryptorchidism torsion after surgery.Among 92 cases of testicular torsion in scrotum,twisted testicles were removed in 41 cases and the testicles were retained in 51 cases;while testicles were removed in 1 case of cryptorchidism torsion,and the testicles were fixed in the scrotum in other 2 cases of cryptorchidism.Sixty eight patients were postoperatively followed-up for 1 month to 15 years,the retained necrotic testis atrophy occurred in 1 case,and the testicular size was normal in other 67 cases.In the 68 cases,there was no testicular discomfort and no testicular torsion after operation.Conclusions CDFI is valuable in preoperative diagnosis of testicular tortion.Once the diagnosis is made,it should be operated as soon as possible to retain the testicular function,and the affected testicle should be retained with caution for cryptorchidism torsion.

Objective To analyze the levels of plasma procalcitonin (PCT)and C-reactive protein (CRP) in 65 patients with septic shock.Methods A total of 65 severe sepsis patients (septic shock group) were selected in our hospital in ICU.After treatment,12 patients died (death group),and 53 cases survived (survival group).At the same time,63 patients with non septic shock were selected (control group).The PCT and CRP levels of the septic shock group and control group were compared and analyzed before and at 3,6 and 12 d after treatment.Results Before treatment,PCT,CRP levels in septic shock group were significantly higher than that in the control group (P ＜ 0.05).At 3,6,12 d of treatment,septic shock group was higher than the control group,the death group was higher than the survival group (P ＜ 0.05).Conclusion The levels of PCT and CRP are important in diagnosis of septic shock progress.

Objective To analyze the levels of plasma procalcitonin (PCT)and C-reactive protein (CRP) in 65 patients with septic shock.Methods A total of 65 severe sepsis patients (septic shock group) were selected in our hospital in ICU.After treatment,12 patients died (death group),and 53 cases survived (survival group).At the same time,63 patients with non septic shock were selected (control group).The PCT and CRP levels of the septic shock group and control group were compared and analyzed before and at 3,6 and 12 d after treatment.Results Before treatment,PCT,CRP levels in septic shock group were significantly higher than that in the control group (P ＜ 0.05).At 3,6,12 d of treatment,septic shock group was higher than the control group,the death group was higher than the survival group (P ＜ 0.05).Conclusion The levels of PCT and CRP are important in diagnosis of septic shock progress.

AIM:To construct AFT024-SCF cell line and HPC-Lhx2 cell line for confirming the biological function of AFT024-SCF.METHODS:The HPC-Lhx2 cell line, AFT024-SCF cell line and AFT024-GFP cell line were constructed by retro-viral infection.The expression of stem cell factor(SCF) in AFT024-SCF cells was detected by real-time PCR and Western blot.SCF in the supernatant of AFT024-SCF was detected with ELISA.The supernatant of AFT024-SCF and AFT024-GFP were collected and then diluted (1:10) with basic IMDM medium.So we made 4 culture medium:AFT024-SCF medium was used for experiment group, AFT024-GFP medium was used for endogenous negative control, IM-DM basic medium was used for exogenous negative control, and IMDM basic medium with SCF was used for positive con-trol.SCF-dependent HPC-Lhx2 cell line was cultured in these 4 different medium for 72 h.According to MTT method and colony forming experiment, the biological function of AFT024-SCF was confirmed by the proliferation ability of SCF-depend-ent HPC-Lhx2 cell line.RESULTS:SCF was highly expressed in AFT024-SCF cells.After cultured for 72 h, neither IM-DM basic medium nor GFP-AFT024 medium support HPC-Lhx2 cell line proliferation.However, AFT024-SCF medium supported HPC-Lhx2 cell line expansion as well as the positive control medium.CONCLUSION:AFT024-SCF cells ex-press SCF successfully and recombinant SCF can be replaced by the supernatant of AFT024-SCF culture medium for expan-ding HPC-Lhx2 cell line in vitro.