Objective To explore the effect of scalp acupuncture combined with repetitive transcranial magnetic stimulation(rTMS)on global cognitive function in patients with post-stroke cognitive impairment(PSCI).To explore the resting-state neuroimaging mechanism of scalp acupuncture combined with rTMS in improving cognitive function in patients with PSCI using fraction amplitude of low frequency fluctuation(fALFF).Methods PSCI patients were randomly assigned to scalp acupuncture group and scalp acupuncture combined with rTMS group(the combined group).Both groups were given conventional rehabilitative training.The scalp acupuncture group selected the affected-side anterior oblique parietotemporal line(MS6).Low frequency(1Hz)rTMS over primary motor cortex(M1)of the unaffected-side in the combined group.The duration was once a day for 20 min,6 sessions one week,in total of 12 sessions for consecutive two weeks.Comparing the total and subtests of the Montreal Cognitive Assessment Scale(MoCA)before and after treatment.At the same time,the statistical method of Difference-in-Difference(DID)was used to analyze the trend of each observational index.The fALFF was used to detect the changes of spontaneous neurological activity in local brain areas and partial correlation analysis was performed with statistical differences cognitive behavioral indicators and significant fALFF values.Results A total of 47 subjects entered the final statistical analyses,including 25 subjects in the scalp acupuncture group and 22 subjects in the combined group.Compared with scalp acupuncture group,there were no significant differences in the global cognitive function in the combined group(P=0.914＞0.05).However,the MoCA subscore-delayed memory task score was significantly improved in the combined group(P=0.035＜0.05).At the same time,the trend analysis also showed that the improvement effect in the combined group was significantly better than those in the scalp acupuncture group(DID=0.80,t=2.20,P=0.033＜0.05).After two weeks of intervention,compared with the baseline fALFF value,the increase of left precuneus,right cerebelum,left fusiform gyrus,left superior parietal gyrus in the combined group were significantly decreased than those of scalp acupuncture group.(GRF corrected,voxel P＜0.01,cluster P＜0.05).The results of partial correlation analysis showed that the fALFF difference of left fusiform gyrus was positively correlated with the difference of subscore-delayed memory in combined group(r=0.719,P＜0.05).Other correlation analyses were not statistically significant(P＞0.05).Conclusion Scalp acupuncture combined with rTMS intervention have definite efficacy in delayed memory.The underpinning neuroimaging mechanism may be related to local neurological spontaneous activity in brain regions involving in cognitive processing such as left fusiform gyrus.

Objective To explore the effect of scalp acupuncture combined with repetitive transcranial magnetic stimulation(rTMS)on global cognitive function in patients with post-stroke cognitive impairment(PSCI).To explore the resting-state neuroimaging mechanism of scalp acupuncture combined with rTMS in improving cognitive function in patients with PSCI using fraction amplitude of low frequency fluctuation(fALFF).Methods PSCI patients were randomly assigned to scalp acupuncture group and scalp acupuncture combined with rTMS group(the combined group).Both groups were given conventional rehabilitative training.The scalp acupuncture group selected the affected-side anterior oblique parietotemporal line(MS6).Low frequency(1Hz)rTMS over primary motor cortex(M1)of the unaffected-side in the combined group.The duration was once a day for 20 min,6 sessions one week,in total of 12 sessions for consecutive two weeks.Comparing the total and subtests of the Montreal Cognitive Assessment Scale(MoCA)before and after treatment.At the same time,the statistical method of Difference-in-Difference(DID)was used to analyze the trend of each observational index.The fALFF was used to detect the changes of spontaneous neurological activity in local brain areas and partial correlation analysis was performed with statistical differences cognitive behavioral indicators and significant fALFF values.Results A total of 47 subjects entered the final statistical analyses,including 25 subjects in the scalp acupuncture group and 22 subjects in the combined group.Compared with scalp acupuncture group,there were no significant differences in the global cognitive function in the combined group(P=0.914＞0.05).However,the MoCA subscore-delayed memory task score was significantly improved in the combined group(P=0.035＜0.05).At the same time,the trend analysis also showed that the improvement effect in the combined group was significantly better than those in the scalp acupuncture group(DID=0.80,t=2.20,P=0.033＜0.05).After two weeks of intervention,compared with the baseline fALFF value,the increase of left precuneus,right cerebelum,left fusiform gyrus,left superior parietal gyrus in the combined group were significantly decreased than those of scalp acupuncture group.(GRF corrected,voxel P＜0.01,cluster P＜0.05).The results of partial correlation analysis showed that the fALFF difference of left fusiform gyrus was positively correlated with the difference of subscore-delayed memory in combined group(r=0.719,P＜0.05).Other correlation analyses were not statistically significant(P＞0.05).Conclusion Scalp acupuncture combined with rTMS intervention have definite efficacy in delayed memory.The underpinning neuroimaging mechanism may be related to local neurological spontaneous activity in brain regions involving in cognitive processing such as left fusiform gyrus.

BackgroundThe incidence of delirium in critically ill psychiatric patients is high， and there are many factors affecting delirium occurrence. At present， epidemiological studies on delirium among critically ill patients in psychiatric hospitals are limited. ObjectiveTo explore the risk factors for delirium in critically ill patients in a psychiatric hospital， so as to guide the clinical management of delirium in psychiatric hospitals. MethodsThis retrospective study included 427 critically ill patients who were admitted to Shenzhen Kangning Hospital from January 1， 2019 to May 31， 2021. The delirium situation， gender， age， pre-admission course of illness （duration from the onset of acute mental state changes to in-patient registration at a psychiatric hospital）， history of mental illness， history of cognitive dysfunction， history of using psychoactive substances， history of using sedative and hypnotic drugs， number of combined chronic diseases， number of combined drugs and type of disease were examined as potential risk factors for delirium. Single Logistic regression was used to analyze the potential risk factors for delirium， and the potential risk factors were incorporated into the multi-factor Logistic regression analysis model so as to gradually screen out the risk factors for delirium in critically ill psychiatric patients. ResultsDelirium was present in 33.49% （143/427） of critically ill patients. Multi-factor Logistic regression analysis demonstrated that the presence of delirium was associated with mental and behavioral disorders caused by psychoactive substances （OR=8.949， P<0.01）， absent history of mental illness （OR=4.202， P<0.01）， number of combined chronic diseases （OR=1.249， P<0.01）， age （OR=1.031， P<0.01） and pre-admission course of illness （OR=0.942， P<0.01） . ConclusionDelirium was present in nearly 1/3 critically ill patients in the psychiatric hospital. The risk factors for delirium included short course of illness before admission， age， more combined chronic diseases， absent history of mental illness， mental and behavioral disorders caused by psychoactive substances. ［Funded by Shenzhen Fund for Guangdong Provincial High-level Clinical Key Specialties （number， SZGSP013）］

Objective Apply modified bilateral carotid artery ligation to establish a VD rat model to observe changes in cerebral blood flow and expression of angiogenic proteins.Methods Thirty-six SD male rats were randomly divided into a sham group(n = 18)and model group(n = 18).In the sham group,only the bilateral carotid artery was isolated without ligation,whereas in the model group,the bilateral carotid artery was ligated to establish the VD model.The Morris water maze behavior test was applied before and 14 days after modeling.Variation in cerebral blood flow was detected by laser speckle contrast imaging.Protein expression of HIF-1α,VEGF,and HO-1 was detected by Western Blot.IL-4 and IL-10 contents were measured by ELISA.Results At 14 days after modeling,escape latency was significantly prolonged and the frequency of crossing the platform had significantly decreased in the model group compared with the sham group(P<0.05).At 2 hours,3 days,and 7 days after modeling,cerebral blood flow in the model group was significantly lower than that in the sham group(P<0.05).At 14 and 21 days after modeling,no significant difference was found in cerebral blood flow between sham and model groups(P>0.05).In the model group,cerebral blood flow was decreased to a minimum at 2 hours after modeling(P<0.05)and then began to recover.The peak of recovery occurred at 3～7 days after modeling and returned to the level before modeling on day 14 after modeling.At postoperative day 21,expression of HIF-1α,VEGF,and HO-1 proteins in the hippocampus of the model group was increased remarkably(P<0.05)and the serum contents of IL-4 and IL-10 in the model group were significantly increased compared with those in the sham group(P<0.05).Conclusions The variation in cerebral blood flow in the VD rat model established by the modified bilateral carotid artery ligation was dependent on time.At postoperative day 21,HIF-1α,VEGF,and HO-1 in the hippocampus were increased significantly,which was accompanied by increased levels of IL-4 and IL-10.

OBJECTIVE: To explore the serological feature and molecular mechanism for a case with A307 subgroup of the ABO blood group system. METHODS: Serological assay was carried out to determine the ABO blood group of the proband and his family members. Genotypes for exons 1 to 7 of the ABO gene were determined with sequence-specific primer polymerase chain reaction (SSP-PCR) and direct sequencing. The impact of the variant on the stability of alpha-1,3-N-acetylgalactosaminyltransferase (GTA) was predicted through construction of a 3D molecular model. RESULTS: The proband, his brother and daughter were diagnosed with Aend phenotype by serological analysis. Their ABO genotype was determined as A307/O02, with heterozygous c.467C>T (p.P156L) and c.745C>T (p.R249W) variants identified in exon 7 of the ABO gene. Molecular modeling suggested that the p.R249W variant may alter the number of hydrogen bonds between the amino acids. The protein was predicted to have a decreased Δ Δ G value of thermodynamic stability. CONCLUSION The p.R249W variant may give rise to the A307 subgroup by reducing the stability of the GTA enzyme, leading to serological features of Aend phenotype.

Clustered regular interspaced short palindromic repeats (CRISPR) system has been widely used in recent years. Compared with traditional genome editing technology, CRISPR/Cas system has notable advantages, including high editing efficiency, high specificity, low cost and the convenience for manipulation. Type Ⅱ and Ⅴ CRISPR/Cas system only requires a single Cas9 protein or a single Cpf1 protein as effector nucleases for cutting double-stranded DNA, developed as genome editing tools. At present, CRISPR/Cas9 technology has been successfully applied to the genome editing of eukaryotes such as zebrafish, mice and human cells, whereas limited progress has been made in the genome editing of bacteria. In our review, we describe CRISPR/Cas system, its mechanism and summarize the optimization and progress of genome editing in bacteria.
Animals ; Bacteria ; CRISPR-Cas Systems ; Endonucleases ; Gene Editing ; Humans ; Mice

OBJECTIVETo explore the molecular mechanism of a case of ABO discrepancies based on the results of blood group serology.

METHODSFive cases of the two-generation pedigrees were analyzed. ABO genotypes were determined using serological tests. DNA sequence analysis was performed on exon 6, exon 7 and intron 3 of the 5 cases to confirm the genotypes of a proband with B subgroup and 4 family members.

RESULTSThere were 3 cases of subgroup AB3 and 1 case of subgroup B3 among the 5 family members. The genotypes were identified as A102/B303 and O02/B303, respectively. B303 differed from B101 by intron 3 point mutation (intron3 + 5G>A).

CONCLUSIONThe point mutation of intron 3 (intron 3+5G>A) is specific in B303.

Objective To discuss the clinical value of protein induced by vitaminK absence antagonist-Ⅱ (PIVKA-Ⅱ)and al-pha-fetoprotein (AFP)in diagnosing primary hepatocellular carcinoma (PHC).Methods There were 178 samples from in-patients in Fuzhou Infectious Disease Hospital,including 54 patients with PHC,39 patients with liver cirrhosis,55 patients with hepatitis and 30 cases of healthy.Serum levels of PIVKA-II and AFP levels were detected by LUMI-PULSEG1200 au-tomatic immunity analyzer and Abbott automatic immunity analyzer respectively,and the difference between the levels was compared.Analyzed the areas under the receiver operating characteristic curves (ROC-AUC)and compared the sensitivity and specificity of single PIVKA-II or AFP assay,and the combined detection of PHC.Results The serum level of PIVKA-Ⅱ in hepatocellular carcinoma group was 274 mAU/ml,which was higher than that in liver cirrhosis group (23 mAU/ml), chronic hepatitis group (26 mAU/ml)and healthy group (21 mAU/ml)(P<0.001),and the levels of AFP in PHC group was 84.0 ng/ml,which was higher than that in liver cirrhosis (21.78 ng/ml)and healthy groups (2.8 ng/ml).But it was not statistically significant (P=0.585)compared with those in the chronic hepatitis group (66.8 ng/ml),the results of re-ceiver operating characteristic (ROC)curve showed that the area under the curve of PIVKA-Ⅱ was 0.776,higher than the AFP (0.649),(Z=2.262,P=0.023 7).Serum PIVKA-Ⅱ (≥40 mAU/ml)had a sensitivity of 78.52% and a specificity of 76.23% in the diagnosis of PHC,While serum AFP (≥10 mg/ml)had a sensitivity of 77.78% and a specificity of 34.64%in the diagnosis of PHC.A combination of serum levels of PIVKA-Ⅱ and AFP could increase the sensitivity in the diagnosis of PHC (vs PIVKA-Ⅱ,P=0.031;vs AFP,P=0.016)and specificity (vs PIVKA-Ⅱ,P=0.004;vs AFP,P=0.001).Con-clusion Serum PIVKA-Ⅱ have high clinical application values in diagnosing PHC.A combination of serum levels of PIV-KA-Ⅱ and AFP could increase the sensitivity and specificity in diagnosis of PHC.

Objective To explore the clinical application of DKK-1,TFF3 and CA72-4 detection in the diagnosis and treatment of gastric cancer.Methods Seventy-five cases(gastric cancer group) of gastric cancer admitted to our hospital from January 2013 to May 2015 were selected.Seventy cases of benign gastric disease(benign gastric disease group) and 70 persons undergoing the physical examination(healthy control group) were selected as the research subjects.The concentration of CA72-4 in each group was detected by the electrochemiluminescence analyzer.The serum DKK-1 and TFF3 levels in each group were detected by enzyme linked immunosorbent assay(ELISA).The receiver operating characteristic(ROC) curve was drawn for evaluating the diagnostic efficiency of each index in each group.The sensitivity,specificity,positive predictive rate and negative predictive rate of 3 indicators for diagnosing gastric cancer before operation were compared.The concentration change of various indexes after gastric cancer radical resection were compared..Results The concentrations of DKK-1,TFF3 and CA72-4 in the gastric cancer group were significantly higher than those in the benign gastric disease group and healthy control group,the difference was statistically significant(P<0.05).The area under ROC curve of DKK-1,TFF3 and CA72-4 were 0.876(95%CI 0.803-0.950),0.944 4(95%CI 0.894-0.975) and 0.818(95%CI 0.726-0.884) respectively.The sensitivity of CA72-4 was 78.6% and the specificity was 84.3%,the sensitivity of DKK-1 was 93.6% and the specificity was 87.1%,the sensitivity and specificity of TFF3 was 92.4% and 90.1% respectively.The concentrations of DKK-1,TFF3 and CA72-4 after radical resection in the gastric cancer group were significantly reduced,the difference was statistically significant compared with before treatment(P<0.05).Conclusion The detection of DKK-1,TFF3 and CA72-4 has a certain clinical value for the diagnosis of gastric cancer.The combined detection of these 3 indicators is conducive to improve the specificity and sensitivity of gastric cancer diagnosis.

Objective To evaluate the clinical effect and efficacy of endoscopy treatment for isolated gastric varices 1 with tissue glue and metal clips. Metheds The clinical date of 21 patients who treated tissue glue and metal clips were retrospectively analyzed from Jan 2015 to Dec 2016. Results The treatments were completed successfully and reviewed by endoscopy after 1 week, 1 month, 3 months, 6 months. The gastric varices were reduced, and the serious complications of bleeding, embolism were little. Conclusion The endoscopy treatment for isolated gastric varices 1 with tissue glue and metal clips is contributed to clinical effect, and the treatment provides a reference for clinical treatment.