Pyruvate maintains and enhances the pro-inflammatory response of microglia caused by glucose deficiency in early stroke.

Pro-inflammatory microglia mainly rely on glycolysis to maintain cytokine production during ischemia, accompanied by an increase in inducible nitric oxide synthase (iNOS) and monocarboxylate transporter 1 (MCT1). The role of energy metabolism in the pro-inflammatory response of microglia is currently unclear. In this study, we tested the response of microglia in mice after cerebral ischemia and simulated an energy environment in vitro using low glucose culture medium. The research results indicate that the expression levels of iNOS and arginase 1 (ARG1) increase in the ischemic mouse brain, but the upregulation of MCT1 expression is mainly present in iNOS positive microglia. In microglia exposed to low glucose conditions, iNOS and MCT1 levels increased, while ARG1 levels decreased. Under the same conditions, knocking down MCT1 in microglia leads to a decrease in iNOS levels, while overexpression of MCT1 leads to the opposite result. The use of NF-κB inhibitors reduced the expression levels of iNOS and MCT1 in microglia. In summary, our data indicate that pyruvate maintains and enhances the NF-κB regulated pro-inflammatory response of microglia induced by low glucose.

RBP-J deficiency promoted the proliferation and differentiation of CD133-positive cells in both in vitro and in vivo studies.

Although Notch signalling pathway could control the proliferation and differentiation of neural stem cells (NSCs), it is largely unknown about the effect of Notch signalling pathway on the neurogenesis of CD133-positive cells. By using the primary cultured ependymal cells and the transgenic mouse, we found that CD133 immunoreactivity was exclusively localized in the ependymal layer of ventricles; moreover, most CD133-positive cells were co-labelled with Nestin. In addition, recombination signal binding protein J (RBP-J), a key nuclear effector of Notch signalling pathway, was highly active in CD133-positive cells. CD133-positive cells can differentiate into the immature and mature neurons; in particular, the number of CD133-positive cells differentiating into the immature and mature neurons was significantly increased following the deficiency or interference of RBP-J in vivo or in vitro. By using real-time qPCR and Western blot, we found that RBP-J and Hes1 were downregulated, whereas Notch1 was upregulated in the expression levels of mRNAs and proteins following the deficiency or interference of RBP-J. These results demonstrated RBP-J deficiency promoted the proliferation and differentiation of CD133-positive cells. Therefore, we speculated that RBP-J could maintain CD133-positive cells in the characteristics of NSCs possibly by regulating Notch1/RBP-J/Hes1 pathway. It will provide a novel molecular insight into the function of RBP-J as well as facilitate a future investigation of CD133-positive cells with respect to their potential application in neurodegenerative disorder.

Three-Dimensional Visualization for Extended Deep Inferior Epigastric Perforator Flaps.

PURPOSE: The aim of the study was to provide an applied and digital anatomical basis of acquiring extended deep inferior epigastric perforator (DIEP) flaps for clinical use. METHODS: Five formalin-soaked specimens were received red latex injection and dissected by layers. The arteriography using the modified mixture of lead oxide-gelatin was performed on 10 adult cadavers that were serially scanned by a spiral computed tomography. The DIEPs were 3 dimensionally reconstructed by Mimics. RESULTS: The medial row perforators of DIEP arteries are located in the medial 1/third of rectus abdominis muscle, and lateral row perforators in the lateral 1/third of the muscle. The perforators distribute mainly from the upper tendinous intersection of umbilicus to below umbilicus within 8.0 cm, especially 4.0 cm. There are constant diameter 0.8-mm perforators or greater accompanied with nerveswithin this region. The main perforators are shown by fast direct volume rendering (VR) reconstruction method, and 3-dimensional images of DIEPs are acquired by dynamic reconstruction (DR) method. Consecutively, the adjacent perforators can be combined freely and the position and anastomosis of extended branches can be easily observed. The extended DIEP flaps were designed by VR and DR methods. CONCLUSIONS: The DIEPs can obtain large extended perforator flaps accompanied with nerves. The perforator close to the umbilicus should be selected while designing the DIEP flap. The 3-dimensional model of extended DIEP flaps can be established conveniently and intuitively by VR and DR methods of Mimics.

Strategies for selecting perforator vessels for transverse and oblique DIEP flap in male pediatric patients: Anatomical study and clinical applications.

Background: Transverse and oblique deep inferior epigastric artery perforator (DIEP) flaps are widely used in breast, lower extremity, urogenital, head and neck reconstruction. In this report, we present our experience with selecting perforator vessels for transverse and oblique DIEP flaps based on an anatomical study and clinical cases. Materials and methods: A detailed anatomical study of the DIEP flap was carried out using a standardized injection of lead oxide in 10 fresh cadavers. Additionally, 35 male pediatric patients (age 5-12 years) underwent lower extremity reconstruction with a DIEP flap. A transverse DIEP flap was used when the defect template did not exceed zone IV, while an oblique DIEP flap was used when the defect template exceeded zone IV. Results: Perforators located below the umbilicus in zones I and II were rich in transverse anastomoses across the midline of the abdominal wall, which is the basis for the transverse DIEP flap. Perforators lateral to the umbilicus in zone I had true anastomoses with the musculophrenic artery, the morphological basis for the oblique DIEP flap. The DIEP flap design was transverse in 20 patients and oblique in 15. Flap sizes ranged from 8 × 4.5 cm2 to 24 × 9 cm2. One oblique DIEP flap was necrosed totally, and it was repaired by a latissimus dorsi musculocutaneous flap. Conclusion: The transverse DIEP flap design based on the perforator located below the umbilicus in zone I is recommended for small skin and soft tissue defects. We recommend the use of the oblique DIEP flap design based on the perforator lateral to the umbilicus in zone I as an extended flap to reconstruct large tissue defects.

Role of circRNAs in neurodevelopment and neurodegenerative diseases.

With the rapid development of sequencing technology, scientists have been able to study and acquire a better understanding of non-coding RNAs (ncRNAs). Circular RNAs (circRNAs), a unique class of ncRNAs with a special loop structure, have been found to possess modulatory properties with respect to various biological processes, such as interacting with nucleic acids or proteins. In addition to their tissue-specific expression and high conservation across species, circRNAs are abundant and dynamically expressed in the nervous system, especially in nerve synapse, indicating their potential regulation in synaptic plasticity or neuronal disorders. In this review, we discuss the characteristics of circRNAs and their common biological functions, as well as their significant role in neurodevelopment, drug addiction and neurodegenerative diseases, aiming to guide further disease diagnoses and efficient therapy.

The Anatomical and Clinical Significance of the Superior Laryngeal Nerve.

OBJECTIVE: This study summarizes the anatomical features of the superior laryngeal nerve in Chinese to enable the rapid location of the superior laryngeal nerve during an operation. STUDY DESIGN: Retrospective analysis of anatomical data. SETTING: Hangzhou First People's Hospital Affiliated to Nanjing Medical University. METHODS: A total of 71 embalmed human cadavers (132 heminecks) were examined over 3 months. The length and diameter of the internal and external branches of the superior laryngeal nerve and their relationships with different landmarks were recorded. RESULTS: The total length of the internal branch of the superior laryngeal nerve was 23.4 ± 6.9 mm. The length of the external branch of the superior laryngeal nerve was 47.7 ± 11.0 mm. Considering the midpoint of the lower edge of the thyroid cartilage as the starting point and using that edge as a horizontal line, when the entry point is above that line, the external branch of the superior laryngeal nerve can be found within 41.1 mm and at an angle of 57.2°. When the entry point is below the lower edge of the thyroid cartilage, the external branch of the superior laryngeal nerve can be found within 34.0 mm and at an angle of 36.5°. CONCLUSION: The superior laryngeal nerve in Chinese people has distinct anatomical characteristics. This article provides a new method of quickly locating the external branch of the superior laryngeal nerve during the operation, which can reduce the probability of damaging the external branch of the superior laryngeal nerve.

Retracted: Allopregnanolone restores the tyrosine hydroxylase-positive neurons and motor performance in a 6-OHDA-injected mouse model.

AIMS: It has been reported that allopregnanolone (APα) promotes the neurogenesis of the neural progenitor cells (NPCs) in the subventricular zone (SVZ) and prevents the decrease of dopaminergic neurons in 6-hydroxydopamine (6-OHDA)-treated mice by binding to γ-aminobutyric acid A receptor (GABAAR) and then opening voltage-gated L-type Ca2+ channel, but the underlying mechanisms remain elusive. The aim of this study was to explore the possible involvement of GABAAR and calcium/calmodulin-dependent protein kinase II delta 3 (CaMKIIδ3) in this process. METHODS: 6-OHDA-treated mice and primary cultured midbrain cells were administrated with APα and GABAAR antagonist bicuculline (Bic), and the proliferation and differentiation of NPCs, the tyrosine hydroxylase (TH)-positive neurons and their fibers, the expression levels of CaMKIIδ3 and brain-derived neurotrophic factor (BDNF), and motor functions were measured using ELISA, immunohistochemical staining, real-time RT-PCR, Western blot, and behavioral test. RESULTS: Allopregnanolone significantly promoted the phosphorylation of cytoplasmic CaMKIIδ3 and its nuclear translocation by binding to GABAAR, which, in turn, increased the expression levels of BDNF. This may account for the findings that the exogenous APα enhanced the proliferation and differentiation of NPCs, and ameliorated the nigrostriatal system and behavioral performance in 6-OHDA-treated mice. CONCLUSIONS: Allopregnanolone may directly activate GABAAR, which, in turn, enhance the proliferation and differentiation of NPCs via upregulating the expression levels of CaMKIIδ3, and finally contribute to the restoration of dopaminergic neurons in 6-OHDA-treated mice.

Allopregnanolone Modulates GABAAR-Dependent CaMKIIδ3 and BDNF to Protect SH-SY5Y Cells Against 6-OHDA-Induced Damage.

Allopregnanolone (APα), as a functional neurosteroid, exhibits the neuroprotective effect on neurodegenerative diseases such as Parkinson's disease (PD) through γ-aminobutyric acid A receptor (GABAAR), but it has not been completely understood about its molecular mechanisms. In order to investigate the neuroprotective effect of APα, as well as to clarify its possible molecular mechanisms, SH-SY5Y neuronal cell lines were incubated with 6-hydroxydopamine (6-OHDA), which has been widely used as an in vitro model for PD, along with APα alone or in combination with GABAAR antagonist (bicuculline, Bic), intracellular Ca2+ chelator (EGTA) and voltage-gated L-type Ca2+ channel blocker (Nifedipine). The viability, proliferation, and differentiation of SH-SY5Y cells, the expression levels of calmodulin (CaM), Ca2+/calmodulin-dependent protein kinase II δ3 (CaMKIIδ3), cyclin-dependent kinase-1 (CDK1) and brain-derived neurotrophic factor (BDNF), as well as the interaction between CaMKIIδ3 and CDK1 or BDNF, were detected by morphological and molecular biological methodology. Our results found that the cell viability and the number of tyrosine hydroxylase (TH), bromodeoxyuridine (BrdU) and TH/BrdU-positive cells in 6-OHDA-treated SH-SY5Y cells were significantly decreased with the concomitant reduction in the expression levels of aforementioned proteins, which were ameliorated following APα administration. In addition, Bic could further increase the number of TH or BrdU-positive cells as well as the expression levels of aforementioned proteins except for TH/BrdU-double positive cells, while EGTA and Nifedipine could attenuate the expression levels of CaM, CaMKIIδ3 and BDNF. Moreover, there existed a direct interaction between CaMKIIδ3 and CDK1 or BDNF. As a result, APα-induced an increase in the number of TH-positive SH-SY5Y cells might be mediated through GABAAR via Ca2+/CaM/CaMKIIδ3/BDNF (CDK1) signaling pathway, which would ultimately facilitate to elucidate PD pathogenesis and hold a promise as an alternative therapeutic target for PD.

[The anatomy basis and clinical application of the horn fasciocutaneous perforator flaps for the reconstruction of the defects on finger soft tissue].

Objective: To discuss the clinical effect of horn perforator flaps for the treatment of finger soft tissue defect. Methods: ① Lead oxide-gelatin mixture injection and CT scanning were performed in 4 fresh adult cadavers and the data was put into mimics 17.0 to achieve the three-dimensional reconstruction. The regulation and origins of the perforator vessel in proper palmar digital artery was observed. ② From July 2012 to July 2015,9 cases with finger soft tissue defects were treated with horn perforator flaps. The defect areas ranged from 0.6 cm × 0.8 cm to 1.2 cm × 1.5 cm and the flap size ranged from0.8 cm×2.5 cm to 1.6 cm×4.2 cm. We put the head end of flaps at defect side. The horn perforator flap was designed with 1-2 perforator vessel and rotated to cover the defect. The defect at donor site was closed directly. Results: We got 4 symmetric arteries in palmar and back surfaces of fingers,2 for proper palmar digital arteries and 2 for dorsal digital arteries. The proper palmar digital arteries have a larger diameter which is the main source of blood supply for fingers. All 9 flaps survived after the operation. Both the recipient area and donor areas received sound healing. With 2 months to 1.5 years of follow-up visit, the shape, color and luster, texture and function all had good healing. Conclusions: The horn perforator flaps for repair finger soft tissue defect have advantages of flexible design, reliable perforator vessels. The defect at donor site can be closed directly with no skin graft. This method considers both shape and function and is a good way for finger soft tissue defect.

[Clinical application of the adjacent horn shaped perforator fasciocutaneous flap in the trunk area].

OBJECTIVE: To study the anatomy basis for the clinical application of the adjacent horn shaped perforator fasciocutaneous flap for the reconstruction of small and medium-sized defects in the trunk area. METHODS: (1) Ten adult antiseptic cadavers (20 sides) were perfused with red latex. The skin blood supply, line of the blood vessels, branches in accordance with the distribution and crossing were observed. (2) Fifteen cases with defects in the trunk were treated with the adjacent horn shaped perforator fasciocutaneous flaps. The defects size ranged from 5 cm x 5 cm to 13 cm x 13 cm with the size of the flaps ranging from 10 cm x 6 cm to 35 cm x 15 cm. RESULTS: The trunk skin is supplied by mainly 17 groups arteries such as thyrocervical trunk, internal thoracic artery, posterior intercostal arteries, superior epigastric artery, arteria epigastrica inferior, lumbar arteries, and so on. The perforators (diameter > 0.5 mm) numbers are about 20, 40, 24, 6, on the chest, abdomen and perineum, upper back, waist, respectively. All the flaps survived completely with primary healing both on donor and recipient sites. The flaps color, texture, function and appearance were satisfactory during the follow-up period of 1-24 months. CONCLUSIONS: The adjacent horn shaped perforator fasciocutaneous flap should be designed flexibly. The defects in the donor sites could be closed directly without skin graft. It is an effective, easy and ideal method for the reconstruction of large defects in the trunk.

[Vascular supply of intrinsic muscles of foot and anatomic basis for muscular flaps design].

OBJECTIVE: To investigate the vascular supply of intrinsic muscles of foot and anatomic basis for muscular flap design. METHODS: A radiopaque injectate (lead oxide-gelatin mixtures, 26 ml/kg) was injected into 10 fresh cadavers. The dissected regions were photographed and each intrinsic muscles on the foot was removed and radiographed. The number, type, diameter of vascular branches of muscles and their distributions were observed. The area of the vascular territory supplied by each source vessel was calculated using Scion Image for Windows software. RESULTS: There were significant architectural differences among the intrinsic muscles. The muscles length varied from 22.5mm to 116.2mm [average, (66.1 +/- 23.2)mm]. The measured fiber length were relatively consistent, ranging from 14.2 mm to 27.5 mm [average, (20.2 +/- 4.5)mm]. There are 63 vascular branches into the 23 foot muscles, each muscle having average branches of 3.2 +/- 0.8. The average diameter of branches, the length and width of each vascular territorial area is (0.8 +/- 0.3) mm, (2.2 +/- 0.8) cm, and (0.9 +/- 0.4) cm, respectively. Other findings included that some muscles were not present in some cadavers. CONCLUSIONS: The blood supply of intrinsic muscles of foot is abundant with different diameter and distributions of branches. There is an anatomic basis for muscular or musculoosseous flap design. There are 7 intrinsic muscles with large and reliable vascular supply which can be chosen as muscular flaps.

[Anatomic study of peroneal tendofascial flap combined with adipofascial flap for the repair of heel tissue defects].

OBJECTIVE: To study the anatomy of peroneal tendofascial flap combined with adipofascial flap for the repair of heel tissue defects. METHODS: The lower extremities of five cadavers (10 sides) were perfused with red latex, the blood supply of peroneal tendofascial flap and vicinity adipofascial flap were observed. The diameter, course, branches and location of the blood vessels were measured. Eight fresh cadavers (16 sides) were perfused with lead oxide-gelatine mixture. The covering fascia tissues of the lower extremities was obtained and photographed by X-ray. The vascular anastomosis and association of nutrient vessel of peroneal tendofascial flap and vicinity adipofascial flap were observed. Two adult lower extremities specimens (4 sides) were used to construct vessel diagrams for observation of the course, distribution and anastomosis of the vessels. Eight cases were treated successfully with theses flaps. RESULTS: The blood supply of the combined fascial flap is multi-originated. For the area within 4 cm below and above the lateral malleolus cusp, the blood supply includes 2-5 branches from heel lateral artery with an average diameter of (0.5 +/- 0.2) mm, 1-2 branches from posterior lateral malleolus artery with an average diameter of (0.6 +/- 0.2) mm and 2-3 branches from the descending part of perforating branches of peroneal artery with an average diameter of (0.5 +/- 0.2) mm. The blood supply of area 4 cm above lateral malleolus cusp is 1-3 branches from intermuscular septum perforating branches of peroneal artery with an average diameter of (1.0 +/- 0.2) mm. These above branches are anastomosed each other and also send off many smaller branches to form vascular net around tendon. The fascial flaps and free skin grafts in eight patients were completely survived. All patients were followed up for 3-24 months, the donor and recipient sites were healed very well. The functional and cosmetic results were satisfactory. CONCLUSIONS: Peroneal tendofascial flap combined with adipofascial flap, with proximal pedicle or reverse distal pedicle, can be used to repair the defect at the lower leg and refractory small- and medium-sized defects at the heel.

[Study on laws of clinical and basic integration acupuncture].

OBJECTIVE: To explore effective ways of integrated clinical and basic studies. METHODS: Try to sum up clinically basic laws of acupuncture from the study thinking of clinical and basic integration. RESULTS: Some laws of clinical acupuncture were raised based on classical theories of the channel and a great of clinical experiences, in combination with basic studies of modern neuroanatomy and physiology. CONCLUSION: The clinical symmetry-corresponding law, the segmental innervation law, the central-axial law, the distal limb dominance law, the law of sallow-deep correlation and the law of local feedback confirm with classical theories of the channels and clinical experiences.