Understanding the Relationship Between Vascular Smooth Muscle Cell Function and the Efficacy of Acupuncture in Treating Cerebral Ischemic Stroke: A Preclinical Meta-Analysis and Systematic Review.

Background: Cerebral blood flow and vascular structures serve as the fundamental components of brain metabolism and circulation. Acupuncture, an alternative and complementary medical approach, has demonstrated efficacy in treating cerebral ischemic stroke (CIS). Nevertheless, the mechanisms underlying the impact of acupuncture on vascular smooth muscle cell (VSMC) function remain uncertain. The objective of this systematic review and meta-analysis is to assess the alterations in VSMC function following acupuncture stimulation in CIS models. Methods: The databases PubMed, Web of Science, SCOPUS, and EMBASE were queried until November 2022 using a predetermined search strategy. The FORMAT BY SYRCLE guidelines were adhered to, and the risk of bias of the included studies was evaluated using the Risk of Bias tool developed by the Systematic Review Centre for Laboratory Animal Experimentation. The random-effects model was employed to estimate the standardized mean difference (SMD). Results: Eighteen articles are included in this review. Acupuncture showed significant positive effects on the region cerebral blood flow (SMD=8.15 [95% CI, 4.52 to 11.78]) and neurological deficiency (SMD=-3.75 [95% CI, -5.54 to -1.97]). Descriptive analysis showed a probable mechanism of acupuncture stimulation in CIS rats related to VSMC function. Limitations and publication bias were presented in the studies. Conclusion: In this systematic review and meta-analysis, our findings indicate that acupuncture stimulation has the potential to improve regional cerebral blood flow and alleviate neurological deficits, possibly by regulating VSMC function. However, it is important to exercise caution when interpreting these results due to the limitations of animal experimental design and methodological quality.

[Effect of electroacupuncture on neurological function and Wnt signaling pathway in ischemic brain tissue of cerebral infarction rats].

OBJECTIVE: To explore the mechanism of electroacupuncture (EA) underlying improvement of cerebral infarction (CI) by investigating its influence on expression of cerebral Wnt7a, lymphoid enhancer factor-1 (LEF1), glycogen synthase kinase 3ß(GSK-3ß) and Dickkopf-1(DKK1) mRNA and proteins in CI rats. METHODS: A total of 280 male Wistar rats were randomly divided into blank control (n＝10), sham-operation, model and EA groups，and 90 rats of the last 3 groups were further divided into 1, 3, 6, 9, 12 and 24 h, and 3, 7 and 12 d subgroups with 10 rats in each subgroup. The CI model was established by occlusion of the middle cerebral artery (MCAO). The sham-operation group received the same surgical operation but without thread embolus insertion. EA (2 Hz/15 Hz, 2 mA) was applied to "Shuigou" (GV26) for 20 min, once a day for 1, 3, 7 and 12 d, respectively. The neurological deficit was evaluated by using Neurological Severity Scores (NSS). The expression levels of Wnt7a，LEF1, GSK-3ß and DKK1 mRNAs and proteins in the right ischemic brain tissues were detected by Quantative real-time PCR and Western blot, respectively. RESULTS: After MCAO, the NSS score was significantly increased in the model and EA groups relevant to the blank control and sham-operation groups (P<0.01) and gradually decreased with the prolongation of ischemia time. After EA, the NSS scores were notably decreased on day 3, 7 and 12 in the EA group compared with the model group (P<0.05, P<0.01). After modeling, the expression levels of Wnt7a and LEF1 mRNAs from 3 h to 12 d, Wnt7a and LEF1 proteins from 6 h to 12 d were considerably increased (P<0.01, P<0.05), while those of GSK-3ß mRNA at 9, 12 and 24 h, GSK-3ß protein at 24 h and 3 d, and DKK1 mRNA at 24 h and 3 d and DKK1 protein at 3 d were obviously decreased in the model group relevant to the sham-operation group (P<0.05, P<0.01). After the intervention, the expression levels of Wnt7a mRNA at 12 h to 3 d, Wnt7α protein from 24 h to 12 d, LEF1 mRNA from 24 h to 12 d, and LEF1 protein from 3 d to 12 d were further apparently up-regulated (P<0.01, P<0.05), while those of GSK-3ß mRNA at 9 h, 3，7 and 12 d, and GSK-3ß protein at 12 h, 7 d and 12 d, and DKK1 mRNA at 12 h, 24 h and 3 d, and DKK1 protein at 24 h to 12 d were obviously down-regulated in the EA group relevant to the model group (P<0.01, P<0.05). No significant difference was found between the blank and sham-operation groups in the NSS scores and expression levels of Wnt7a, LEF1, GSK-3ß and DKK1 mRNAs and proteins at all the time points (P>0.05). CONCLUSION: EA of GV26 can significantly improve the neurological deficit symptoms in MCAO rats, which may be associated with its effects in up-regulating the expression of Wnt7a and LEF1 mRNAs and proteins, and in down-regulating the expression of GSK-3ß and DKK1 mRNAs and proteins.

[Effect of manual acupuncture of "Shuigou" (GV26)"Neiguan" (PC6) on neurological function and expression of apoptosis-related factors in brain tissues surrounding hematoma in intracerebral hemorrhage rats].

OBJECTIVE: To observe the effect of manual acupuncture stimulation of "Shuigou" (GV26) and "Neiguan" (PC6) on neurological function and expression of Caspase-3 and Caspase-9 in brain tissues around the intracerebral hematoma in rats with acute intracerebral hemorrhage (ICH), so as to explore its possible mechanisms underlying improvement of ICH. METHODS: Ninety-six male SD rats were randomly divided into 4 groups: control, model, acupoint and non-acupoint (24 rats in each group). The ICH model was established by injection of the rat's autologous blood into the caudate nucleus. According to the time-points of 6, 24, 48 and 72 h after ICH, each of the 4 groups was further divided into 4 subgroups. For rats of the acupoint group, the PC6 on both sides was manually stimulated by manipulating the needle with lifting-thrusting-twisting reducing techniques, while the GV26 was stimulated with strong "sparrow-pecking" method for 10 times, then, left the needles in the acupoints for 30 min. For rats of the non-acupoint group, two non-acupoints: mid-spot below the bilateral axilla and the spot 3 mm above the left side of the coccyx tip were stimulated with the same methods to PC6 and GV26, respectively. For rats of the 6 h and 24 h subgroups, the intervention was given once after waking up from modeling, and for those of the 48 and 72 h subgroups, the intervention was conducted once a day for 2 or 3 times, respectively. The neurological severity score (NSS) was used to evaluate the degree of neurological function. The immunoactivity (expression) of Caspase-3 and Caspase-9 proteins of the hematoma focus of the brain was detected by immunohistochemistry. RESULTS: Following modeling, the NSS and the expression levels of Caspase-3 and Caspase-9 proteins in the brain tissues surrounding the hematoma at each time-points (6, 24, 48 and 72h) after modeling were significantly increased in the model group relevant to the control group (P<0.01, P<0.05). Compared with the model group, the NSS at 72h and the expression levels of Caspase-3 and Caspase-9 proteins at 6, 24, 48 and 72h were significantly down-regulated in the acupoint group (P<0.05) rather than in the non-acupoint group (P>0.05). CONCLUSION: Acupuncture of GV26 and PC6 can improve the neurological function in rats with ICH, which may be related to its function in reducing the expression of Caspase-3 and Caspase-9 proteins (apoptosis-related proteins) in the brain.

[Treatment of growing Class II division 1 patients with varying degrees of anteroposterior and vertical dysplasias].

PURPOSE: To evaluate the difference of treatment on growing patients with Class II division 1 using various anteroposterior and vertical skeletal dysplasias. METHODS: Twenty-eight moderate to severe Class II division 1 malocclusion patients with Class I molar and canineocclusion after treatment were selected. The subjects were divided into 3 groups based on the initial ANB and FMA angles. Pre-treatment and post-treatment cephalograms were evaluated for soft and hard tissues changes after treatment. The data was analyzed using SPSS 12.0 software package. RESULTS: ANB angle decreased in all three groups. Changes of ANB angle in group 2 and group 3 were greater than group 1(P<0.01). SNA angle decreased significantly in group 3(P<0.01). SNB angle increased significantly in group 1(P<0.01) and group 2(P<0.05). All groups had great maxillary incisors retraction and similar changes in mandibular incisor positions.Soft tissue index showed no significant difference among three groups before and after treatment(P>0.05). But nasolabial angle(P<0.05), soft tissue convexity(P<0.05) and Z angle(P<0.01) changed significantly in group 3. CONCLUSIONS: Conventional orthodontic therapy successfully corrects Class II division 1 malocclusions in growing patients with the improvement of soft and hard tissues.The patients who initially have the most severe skeletal dysplasias can have the greatest changes in profile.

[Non-extraction treatment in borderline class III malocclusion].

PURPOSE: To evaluate the effect of non-extraction treatment protocol combined with rapid palatal expansion and lower molar distalization in borderline Class III malocclusion in adolescents. METHODS: Nine borderline Class III patients (average age: 13.4 ± 0.7 years) with average (6.7 ± 2.8)mm crowding in maxilla and (2.6 ± 0.4) mm crowding in mandible received the treatment described above. The lateral cephalograms of these patients were analyzed. All patients had a pretreatment ANB angle greater than -3°. The difference before and after treatment was analyzed with paired t test using SPSS11.0 software package. RESULTS: Evaluation of cephalometric measurements revealed forward movement of the maxilla, indicated by 1.69 mm forward positioning of A-point (P < 0.05), and an increase in SNA by 1° (P = 0.054). The mandible was inhibited from growing forward with slight retrusion evidenced by a decreased B-point (-1.88 mm, P < 0.05), and a decrease in SNB by 0.69° (P < 0.05). The combined maxillary and mandibular changes resulted in an increase in ANB angle (1.69°, P < 0.05). A clockwise rotation of the mandible was found, with an increase in Y-axis (0.88°,P < 0.05) and SN-MP (1.50°,P<0.05). Significant change was also found in the forward movement of maxillary incisors (+3.44 mm, P < 0.05), and the slight retrusive positioning of mandibular incisors (-0.5 mm, P > 0.05). An increase in U1-SnPg' (1 mm, P < 0.05) and a decrease in L1-SnPg' (0.63 mm, P > 0.05) resulted in the improvement of upper and lower lips relationship (1.88 mm, P < 0.05). CONCLUSION: This treatment protocol can improve the sagittal jaws and upper-lower lip relationship.

[A comparison of rapid palatal expansion and Damon appliance on non-extraction correction of dental crowding].

PURPOSE: To compare the effect of rapid palatal expansion (RPE) and Damon technique on the correction of dental crowding with non-extraction approach. METHODS: 19 adolescent patients with non-extraction orthodontic treatment were reviewed. They had Class I skeletal pattern and moderate crowding. Nine subjects were treated with RPE followed by straight wire appliance, and 10 subjects were treated with Damon appliance only. The dental casts and cephalometric radiophotographs were measured before and after the treatment. The difference between two groups was analyzed for Student's t test with SPSS15.0 software package. RESULTS: Upper and lower inter-canine width increased 2.9mm and 1.3mm respectively in RPE group (P<0.05), while there was no significant change in Damon group. A significant posterior expansion of upper and lower arch width was noted in both RPE and Damon groups (P<0.05). The increase in upper inter-bicuspid width and lower inter-molar width was comparable in two groups. The upper inter-molar width in RPE group expanded 5.3mm, which was significantly larger than that of 3.1mm in Damon group(P<0.05). The increase in lower inter-bicuspid width in Damon group, on the other hand, was larger than that of RPE group (4.5mm vs. 1.5mm). (P<0.05) Both groups showed buccal tipping of bicuspids and molars, with more extend of bicuspid tipping in Damon group (P<0.05). The maxillary base width increased 2.1mm in RPE group, which was significantly larger than that of 0.6mm in Damon group (P<0.05). The upper and lower incisors in Damon group were tipped forward by 9.2 degrees and 7.5 degrees respectively, while the change in RPE group was not significant (P<0.05). CONCLUSION: Both RPE and Damon technique can successfully increase the arch width and correct moderate dental crowding with non-extraction approach. Damon appliance protrudes the upper and lower incisors and expands the dental arch by buccal tipping of bicuspids and molars. RPE expands the maxillary base along with the whole upper dental arch and could maintain the incisors in upright position.

[The effect of maxillary protraction in early permanent dentition].

PURPOSE: To evaluate the effect of maxillary protration in skeletal anterior crossbite patients in early permanent dentition and determine the optimal timing for orthopedic treatment. METHODS: 18 patients (11 males and 7 females with a mean age of 11.5 years) with concave profile were treated with protraction headgear. For each patient lateral cephalogram was taken before and after treatment. Cephalometric analysis was carried out by using linear and angular measurements. Statistical analysis was performed using SAS6.12 software package. RESULTS: All subjects were treated to normal overjet. SNA was increased by 2.06 degrees (P<0.001) ,A point was protruded by 3.94 mm(P<0.001), U1-SN was increased by 6.97 degrees (P<0.001).SNB was decreased by 1.39 degrees (P<0.01),SN-MP was increased by 2.64 degrees (P<0.001),Y axis was increased by 2.42 degrees (P<0.001), Pg point was decreased by 7.42 mm(P<0.001). Maxillary incisor was promoted by 6.86 mm,which was a result of 57.43% skeletal change and 42.57% dental change. CONCLUSIONS: Maxillary protraction can significantly promote maxillary growth, change mandibular growth direction and inhibit mandibular protrusion in early permanent dentition.

[Preliminary clinical study on intrusion of overerupted molars with a removable appliance].

PURPOSE: To evaluate the clinical effect of molar intrusion with a removable appliance and determine its clinical value. METHODS: Seven patients were selected in the study.The clinical effect of intrusion was evaluated,the periodontal attachments and root resorption post-treatment were accessed by models,clinical examination, periapical radiograph analysis via CygnusRay2TM digital X-ray system pre and post-treatment.The data was analyzed using SAS6.12 software package. RESULTS: All the overerupted molars had been intruded successfully within an average time of 3.7 months.Pre and post-treatment model analysis demonstrated the maximum intrusion amount was 3.05mm,minimum 1.03mm and mean 0.58mm per month.Analysis of pre and post-treatment periapical radiographs and periodontal probing depth showed no significant difference in probing depth,distance from cemento-enamel junction to alveolar bone crest,mesial and distal tooth length between pre- and post- treatment (P>0.05). CONCLUSIONS: Molar intrusion with the removable appliance has a significant effect without apparent destruction of the periodontal tissue and root resorption.

Most spontaneous tumors in a mouse model of Li-Fraumeni syndrome do not have a mutator phenotype.

Mutations are the substrate of cancer. Yet, little is known about the degree and nature of mutations in tumors because measurement of mutation load in tumors and normal tissues was generally not possible until the advent of transgenic mouse mutation detection systems. Herein, we present the first analysis of mutation frequency and pattern in thymic tumors from a mouse model of Li-Fraumeni syndrome (p53+/- murine model) using the Big Blue assay with sequencing of all mutants. We also make the first characterization of mutation frequency and pattern in p53-deficient extra-thymic cancers. The data more than triple the literature on all non-mismatch repair deficient tumors for which mutations are identified by sequence analysis, allowing mutation frequency and pattern to be determined. Most tumors had a normal mutation frequency and a normal mutation pattern. Five tumors showed modest increases in mutation frequency (2.3-fold or less). Alterations in mutation patterns were uncommon, tumor-specific and not necessarily associated with increases in mutation frequency. Given the data from two spontaneous tumors (normal mutation frequency with an abnormal pattern in a p53-/- mouse and low mutation frequency in a p53+/+ control mouse), we hypothesize that tumors sometimes can carry a low mutation load. The study was not without certain caveats: mutation load could not be compared between tumor and normal tissue from the same animal; sample sizes for extra-thymic tumor types were small, and only point mutations and deletions, insertions and indels up to 2 kb were detected. However, the data clearly show key differences in tumors from p53+/- mice compared with mismatch repair deficient tumors; a lack of dramatic increase in mutation frequency and absence of a signature of mutation.

[Novel mechanism of 3' exonuclease of polymerase in maintenance of DNA replication fidelity and its application in SNP assay].

Polymerase with 3' to 5'exonulcease plays an important role in the maintenance of in vivo DNA replication fidelity. In order to develop more reliable SNP assays, we revisit the underlying molecular mechanisms by which DNA polymerases with 3' exonucleases maintain high fidelity of DNA replication. In addition to mismatch removal by proofreading, we recently discovered a premature termination of polymerization by a new mechanism of OFF-switch. This novel ON/OFF switch turns off DNA polymerization from mismatched primers and turns on DNA polymerization from matched primers. Two SNP assays were developed based on the proofreading and the newly identified OFF-switch respectively: terminal labeled primer extension and the ON/OFF switch operated SNP assay. These two new methods are well adapted to conventional techniques such as electrophoresis, real time PCR, microplates, and microarray. Application of these reliable SNP assays will greatly facilitate genetic and biomedical studies in the post-genome era.

Exo+ proofreading polymerases mediate genetic analysis and its application in biomedical studies.

Polymerases with a proofreading function in their internal 3' to 5' exonuclease possess high fidelity for DNA replication both in vivo and in vitro. The obstacle facing Exo+ polymerases for single nucleotide polymorphism (SNP) detection could be bypassed by using primer-3'-termini modification. This hypothesis has been well tested using three types of modified allele specific primers with: 3' labeling, 3' to 5' exonuclease resistance, and 3' dehydroxylation. Accordingly, three new SNP assaying methods have been developed to carry out genome-wide genotyping, taking advantage of the enzymatic properties of Exo+ polymerases. These new mutation detection assays are widely adaptable to a variety of platforms, including multi-well plate and microarray technologies. Application of Exo+ polymerases to genetic analysis, including genotyping that is mostly relevant to pharma-cogenetics, high-fidelity gene expression profiling, rare mutation detection and mutation load assay, will help to accelerate the pace of personalized medicine. In this review paper, we will first introduce three new assays that we have recently developed, and then describe a number of their applications in pharmacogenetics and in other biomedical studies.

[Morphologic characteristics of anterior crossbite in early permanent dentition reviewed by superimposition on the template].

PURPOSE: To establish a computer-processing module to first, eliminate the factors which mislead the diagnosis of anterior crossbite and second, superimpose the individual tracing on the template to review the patient's morphologic characteristics. METHODS: First, the module would process the following steps:1.to create three templates; 2.to move the functional-shift-mandible posteriorly; 3.to standardize the potential of vertical dimension;4.to select a suitable template for superimposition; 5.to standardize the sizes of individual graph and template. Second, the module would process the five-step superimpositions: 1. X axis was superimposed and S were registered to measure the distances between Ptm points and Ar points on X axis; 2. X axis was paralleled and Ptm were registered to measure the distance between A points on X axis; 3. X axis was paralleled and A points were registered to measure the difference of anterior area between the mandibular graphs; 4.mandibular planes were superimposed and Po was registered to show the mandibular form variation; 5. Ar-Gn lines were superimposed and Ar was registered to measure the difference between the mandibular graphs. RESULTS: The module which could complete these steps was produced. With its help, the individual graph could be diagnosed with the position of maxilla and mandible, the amounts of maxillary discrepancy and jaws' dysplasia, the mandibular form and size variation. CONCLUSIONS: The module could eliminate the factors which mislead the diagnosis and review the morphologic characteristics of anterior crossbite in early permanent dentition.

On/off switch mediated by Exo+ polymerases: experimental analysis for its physiological and technological implications.

The potential physiological role and technological application of the premature termination of DNA polymerization through the off-switch of exo+ polymerases were studied using 3' phosphorothioate-modified or unmodified primers with single base mismatch distal to the 3' terminus. With exonuclease-digestible unmodified primers, a gradient premature termination of DNA polymerization was observed when amplified with exo+ polymerases. With 3' allele specific phosphorothioate-modified primers, an efficient off-switch effect occurred in the discrimination of a single nucleotide polymorphism when directly using genomic DNA. Clearly, the off-switch of exo+ polymerases is useful in biomedical research.

[Effect of 3' exonuclease activity of polymerase on extension of phosphorothioate-modified primers].

OBJECTIVE: To determine whether 3'phosphorothioate-modified-2 terminal mismatched primers can turn off DNA polymerization mediated by Exo(+) polymerase. METHODS: Two-directional primer extension was performed using polymerase with and without 3' exonuclease activity. The effects of unmodified primers and 3' phosphorothioate-modified primers on primer extension were evaluated. RESULTS: Exo(-) polymerase yielded products from matched and mismatched primers regardless of their modification. However, 3' phosphorothioate-modified primers with a single base mismatch at -2 position worked similarly to the terminal (-1) mismatched primers in triggering the novelly reported "off-switch" of Exo(+) polymerase. CONCLUSION: These data suggested that the "off-switch" can be of enormous application in the diagnosis of single gene diseases and in the association studies by single nucleotide polymorphism screening.