Infectious meningitis. Why are the leptomeninges preferentially involved? Electron microscopic insights.

In infectious meningitis, pathogens preferentially attack the leptomeninges (pia mater and arachnoid) rather than the pachymeninges (dura mater). This study aims to provide ultra-anatomical insights from our extensive collection of electron microscopy images and propose mechanisms, highlighting structures that favor the introduction, adherence, colonization, and proliferation of microorganisms leading to spinal meningitis. Over several years, we analyzed an extensive collection of transmission and scanning electron microscopy images of human spinal meninges captured in our laboratories. Upon examining 378 of those images, we identified potential sites for the iatrogenic or hematogenic introduction and adherence of microorganisms, as well as sites for their colonization and proliferation. These included the outer surface of the spinal dural sac, structures within the epidural space, and the spinal dural sac itself, which comprises compact dura mater with interwoven collagen fibers and tightly bound arachnoid cells. Also, the subdural (extra-arachnoid) compartment, consisting of fragile neurothelial cells prone to rupture under force, formed an acquired spinal subdural space, a new subarachnoid compartment, limited by arachnoid trabeculae, that surrounded the nerve roots and spinal cord and the pia mater. Macrophages, fibroblasts, mast cells, and plasma cells were also observed within the dura mater, arachnoid layer, arachnoid trabeculae, and pia mater. These images illustrate how the characteristics of the meningeal layers could contribute to bacterial adhesion and proliferation at various locations, inducing selective inflammation during (iatrogenic) spinal meningitis. In addition, the images help to explain why magnetic resonance imaging enhancement appears preferentially at specific sites.

Morphological variability of lateral femoral cutaneous nerve and its potential clinical significance.

BACKGROUND: The lateral femoral cutaneous nerve is derived from the dorsal branches of the L2 and L3 spinal nerves. It travels across the pelvis and heads towards the anterior superior iliac spine. It passes under the lateral part of the inguinal ligament and then divides into two branches, which are responsible for sensory innervation of the anterolateral and lateral skin of the thigh. However, the course of this nerve can vary morphologically. Numerous differences have been observed in its exit from the pelvis and in the number of its main trunks and branches. Additionally, its angle with the inguinal ligament and its placement in relation to other structures (such as the femoral artery, femoral nerve, and the sartorius and iliacus muscles) also vary. All of these variants have potential clinical implications. Therefore, the aim of this review is to present the morphological variability of the lateral cutaneous nerve and to explore how these anatomical differences can introduce clinical concerns. MATERIALS AND METHODS: Presented review of the literature was written based on over 30 studies. Comprehensive literature search was done using PubMed in order to study the morphological variability of lateral femoral cutaneous nerve (LFCN). To be included in this review studies needed to be meet certain criteria: been published before December 2023, present information valuable to this paper (variability of lateral femoral cutaneous nerve/clinical significance). The search included how LFCN vary either among fetuses and adults in the aim of providing more complex information about the variability of this nerve. During the search key words as following were used. No particular references were excluded from the analysis. All relevant studies were included, and citation tracking was used to identify publications. RESULTS: This review presents the description of variability of LFCN and its potential clinical impact. In the review differences in adult and fetuses were considered, morphological variability were divided into 4 groups: the origin of the nerve, the way it leaves the pelvis, the branching pattern, the angle between LFCN and surrounding structures and then, clinical significance were considered basing on available literature.

Possible anastomoses between the long branches of the brachial plexus and their clinical significance.

The brachial plexus consists of nerves that supply the upper limb and some nerves of the back, torso, and neck. It is formed by the ventral rami of C5 to T1 (in some cases, C4 or T2 also contribute). The anterior rami of the spinal nerves unite to the roots, trunks, divisions, cords, and terminal branches that innervate muscles and skin. An example is associated with terminal branches of the long nerves. Knowledge of this variation is necessary for enabling surgeons, orthopedists, and neurologists to avoid injury during surgical exploration in the arm or axilla region, and for achieving correct diagnoses, because such variability can evoke nonspecific responses. Awareness of this anastomosis is also mandatory for anesthetists performing anesthesia in the upper limb region. The aim of this article is to describe anastomoses between long nerves from the brachial plexus and to consider their clinical significance.

The morphological variability of the piriformis muscle.

BACKGROUND: The aim of the study is to create several classifications of the piriformis muscle (PM): proximal and distal attachments, potential fusions, and the relationship with the sciatic nerve. It is the first comprehensive anatomical examination of this subject. MATERIALS AND METHODS: One hundred and twenty-four lower limbs from 62 cadavers, fixed in 10% formalin, were examined. RESULTS: The piriformis muscle was present in 120 limbs (96.8% of cases). Four types of proximal attachment were described (I-IV). The most common type was Type I, in which the proximal attachment was at the anterior surface of the sacrum, between S2 and S4 (52 lower limbs; 43.3%). The rarest type was Type IV, in which the proximal attachment was at the gluteal surface of the ilium near the margin of the greater sciatic notch and from the gluteus medius (12 cases; 10%). Three types of distal attachment were distinguished. The most common was Type 1, a single tendon. This type comprised two subtypes: A and B (105 lower limbs; 87.5%). The other two types accounted for 12.5% of the total. Fusions were noted between the piriformis muscle and adjacent muscles in 31.7%. Four patterns were observed in which the sciatic nerve ran against the piriformis muscle. The most common variation in the relationship was the common fibular nerve exiting superior to the piriformis muscle and the tibial nerve passing inferior to it (10 cases; 8.3%). CONCLUSIONS: The piriformis muscle is highly morphologically variable in both its proximal and distal attachments and its relationship with the sciatic nerve. There are four types of proximal attachment and three types of distal attachment. The piriformis muscle shows numerous fusions with its adjacent muscles: gluteus medius or minimus or superior gemellus. A new (fourth) type of relationship was demonstrated between the piriformis muscle and sciatic nerve. The piriformis muscle was absent in four cases.

Anatomical study with clinical significance of communicating and visceral branching of the cervical and upper thoracic sympathetic trunk.

Current advances in the management of the autonomic nervous system in various cardiovascular diseases, and in treatments for pain or sympathetic disturbances in the head, neck, or upper limbs, necessitate a thorough understanding of the anatomy of the cervicothoracic sympathetic trunk. Our objective was to enhance our understanding of the origin and distribution of communicating branches and visceral cervicothoracic sympathetic nerves in human fetuses. This was achieved through a comprehensive topographic systematization of the branching patterns observed in the cervical and upper thoracic ganglia, along with the distribution of communicating branches to each cervical spinal nerve. We conducted detailed sub-macroscopic dissections of the cervical and thoracic regions in 20 human fetuses (40 sides). The superior and cervicothoracic ganglia were identified as the cervical sympathetic ganglia that provided the most communicating branches on both sides. The middle and accessory cervical ganglia contributed the fewest branches, with no significant differences between the right and left sides. The cervicothoracic ganglion supplied sympathetic branches to the greatest number of spinal nerves, spanning from C5 to T2. The distribution of communicating branches to spinal nerves was non-uniform. Notably, C3, C4, and C5 received the fewest branches, and more than half of the specimens showed no sympathetic connections. C1 and C2 received sympathetic connections exclusively from the superior ganglion. Spinal nerves that received more branches often did so from multiple ganglia. The vertebral nerve provided deep communicating branches primarily to C6, with lesser contributions to C7, C5, and C8. The vagus nerve stood out as the cranial nerve with the most direct sympathetic connections. The autonomic branching pattern and connections of the cervicothoracic sympathetic trunk are significantly variable in the fetus. A comprehensive understanding of the anatomy of the cervical and upper thoracic sympathetic trunk and its branches is valuable during autonomic interventions and neuromodulation. This knowledge is particularly relevant for addressing various autonomic cardiac diseases and for treating pain and vascular dysfunction in the head, neck, and upper limbs.

Are There Any Differences in the Healing Capacity between the Medial Collateral Ligament's (MCL) Proximal and Distal Parts in the Human Knee? Quantitative and Immunohistochemical Analysis of CD34, α-Smooth Muscle Actin (α-SMA), and Vascular Endothelial Growth Factor (VEGF) Expression Regarding the Epiligament (EL) Theory.

The human knee is a complex joint that comprises several ligaments, including the medial collateral ligament (MCL). The MCL provides stability to the knee and helps prevent its excessive inward movement. The MCL also has a thin layer of connective tissue known as the epiligament (EL), which adheres to the ligament. This unique feature has drawn attention in the field of ligament healing research, as it may have implications for the recovery process of MCL injuries. According to the EL theory, ligament regeneration relies heavily on the provision of cells, blood vessels, and molecules. The present study sought to compare the expression of vascular endothelial growth factor (VEGF), CD34, and α-smooth muscle actin (α-SMA) in healthy knees' proximal and distal MCL segments to better understand how these proteins affect ligament healing. By improving the EL theory, the current results could lead to more effective treatments for ligament injury. To conduct the present analysis, monoclonal antibodies were used against CD34, α-SMA, and VEGF to examine samples from 12 fresh knee joints' midsubstance MCLs. We identified a higher cell density in the EL than in the ligament connective tissue, with higher cell counts in the distal than in the proximal EL part. CD34 immunostaining was weak or absent in blood vessels and the EL, while α-SMA immunostaining was strongest in smooth muscle cells and the EL superficial layer. VEGF expression was mainly in the blood vessels' tunica media. The distal part showed more SMA-positive microscopy fields and higher cell density than the proximal part (4735 vs. 2680 cells/mm2). Our study identified CD34, α-SMA, and VEGF expression in the MCL EL, highlighting their critical role in ligament healing. Differences in α-SMA expression and cell numbers between the ligament's proximal and distal parts may explain different healing capacities, supporting the validity of the EL theory in ligament recovery.

Do the Differences in the Epiligament of the Proximal and Distal Parts of the Anterior Cruciate Ligament Explain Their Different Healing Capacities? Quantitative and Immunohistochemical Analysis of CD34 and α-SMA Expression in Relation to the Epiligament Theory.

The aim of this study was to assess the epiligament theory by determining the normal epiligament morphology of the proximal and distal parts of the anterior cruciate ligament in humans and analyzing the differences between them and the midportion of the ligament in terms of cell numbers and expression of CD34 and α-SMA. Samples were obtained from the anterior cruciate ligaments of 12 fresh knee joints. Monoclonal antibodies against CD34 and α-SMA were used for immunohistochemistry. Photomicrographs were analyzed using ImageJ software, version 1.53f. The cell density was higher in the epiligament than in the ligament connective tissue. Cell counts were higher in the proximal and distal thirds than in the midsubstance of the epiligament. CD34 was expressed similarly in the proximal and distal thirds, although it seemed slightly more pronounced in the distal third. α-SMA expression was more robust in the proximal than the distal part. The results revealed that CD34 and α-SMA are expressed in the human epiligament. The differences between the numbers of cells in the proximal and distal parts of the epiligament and the expression of CD34 and α-SMA enhance epiligament theory. Future investigations into improving the quality of ligament healing should not overlook the epiligament theory.

A very rare case report: accessory head of the sartorius muscle.

The sartorius muscle belongs to the anterior compartment of the thigh. Morphological variations of this muscle are very rare, few cases being described in the literature. An 88-year-old female cadaver was dissected routinely for research and teaching purposes. However, an interesting variation was found during anatomical dissection. The proximal part of the sartorius muscle had the normal course, but the distal part bifurcated into two muscle bellies. The additional head passed medially to the standard head; thereafter, there was a muscular connection between them. This connection then passed into the tendinous distal attachment. It created a pes anserinus superficialis, which was located superficially to the distal attachments of the semitendinosus and gracilis muscles. This superficial layer was very wide and attached to the medial part of the tibial tuberosity and to the crural fascia. Importantly, two cutaneous branches of the saphenous nerve passed between the two heads. The two heads were innervated by separate muscular branches of the femoral nerve. Such morphological variability could be clinically important.

Very rare arrangement of the pes anserinus: potential clinical significance.

The pes anserinus superficialis is composed of the semitendinosus, gracilis and sartorius tendons. Normally, they all insert to the medial side of the tibial tuberosity, and the first two are attached superiorly and medially to the tendon of the sartorius muscle. During anatomical dissection, a new pattern of arrangement of tendons creating the pes anserinus was found. The pes anserinus comprised three tendons; the semitendinosus tendon was located superiorly to the gracilis tendon, and they both had distal attachments on the medial side of the tibial tuberosity. This seemed like the normal type, but the tendon of the sartorius muscle created an additional superficial layer, its proximal part lying just below the gracilis tendon and covering the semitendinosus tendon and a small part of the gracilis tendon. After crossing the semitendinosus tendon it is attached to the crural fascia significantly below the tibial tuberosity. Good knowledge of the morphological variations of the pes anserinus superficialis is necessary during surgical procedures in the knee region, especially anterior ligament reconstruction.

Two variant muscles in the gluteal region.

The short lateral rotators of the thigh found in the gluteal region can have morphological variations. During anatomical dissection of a right lower limb, two variant structures were found in this region. The first of these accessory muscles originated from the external surface of the ramus of the ischium. Distally, it was fused with the gemellus inferior muscle. The second structure comprised tendinous and muscular parts. The proximal part originated from the external part of the ischiopubic ramus. It inserted on the trochanteric fossa. Both structures were innervated by small branches of the obturator nerve. The blood supply was via branches of the inferior gluteal artery. There was also a connection between the quadratus femoris and the superior part of the adductor magnus. These morphological variants could be clinically important.

Accessory part of the deltoid muscle.

The shoulder and arm region has numerous morphological variations. The deltoid muscle usually consists of three parts: anterior, middle and posterior. This case report describes a very rare deltoid muscle variant, an addition to the spinal part that is attached proximally at the infraspinatus fascia and the spine of the scapula. The distal attachment transforms directly into the brachialis muscle. Additional parts can affect the biomechanics and function of the joints significantly.

Morphological variability of the leg muscles: potential traps on ultrasound that await clinicians.

BACKGROUND: Although muscles and their tendons are not considered the most morphologically variable structures, they still manifest a substantial diversity of variants. The aim of this study is to increase awareness of some of the many possible variants found during ultrasound imaging of one lower limb compartment, the leg, that could potentially mislead clinicians and lead to misdiagnosis. MATERIALS AND METHODS: PubMed was used for a comprehensive literature search for morphological variations. Relevant papers were included, and citation tracking was used to identify further publications. RESULTS: Several morphological variants of muscles of the leg have been described over many years, but this study shows that the occurrence of further variations in ultrasound imaging requires further investigations. CONCLUSIONS: The incidence of additional structures including muscles and tendons during ultrasound examination can cause confusion and lead to misinterpretation of images, misdiagnosis, and the introduction of unnecessary and inappropriate treatments.

The size, number, and distribution of nerve endings around and within the human epiglottis, focusing on tracheal intubation maneuvers.

The aim of this study was to examine the distribution of nerve endings in the mucosa, submucosa, and cartilage of the epiglottis and the vallecula area and to quantify them. The findings could inform the choice of laryngoscope blades for intubation procedures. Fourteen neck slices from seven unembalmed, cryopreserved human cadavers were analyzed. The slices were stained, and cross and longitudinal sections were obtained from each. The nerve endings and cartilage were identified. The primary metrics recorded were the number, area, and circumference of nerve endings located in the mucosa and submucosa of the pharyngeal and laryngeal sides of the epiglottis, epiglottis cartilage, and epiglottic vallecula zone. The length and thickness of the epiglottis and cartilage were also measured. The elastic cartilage of the epiglottis was primarily continuous; however, it contained several fragments. It was covered with dense collagen fibers and surrounded by adipose cells from the pharyngeal and laryngeal submucosa. Nerve endings were found within the submucosa of pharyngeal and laryngeal epiglottis and epiglottic vallecula. There were significantly more nerve endings on the posterior surface of the epiglottis than on the anterior surface. The epiglottic cartilage was twice the length of the epiglottis. The study demonstrated that the distribution of nerve endings in the epiglottis differed significantly between the posterior and anterior sides; there were considerably more in the former. The findings have implications for tracheal intubation and laryngoscope blade selection and design.

The role of an open artificial intelligence platform in modern neurosurgical education: a preliminary study.

The use of artificial intelligence in neurosurgical education has been growing in recent times. ChatGPT, a free and easily accessible language model, has been gaining popularity as an alternative education method. It is necessary to explore the potential of this program in neurosurgery education and to evaluate its reliability. This study aimed to show the reliability of ChatGPT by asking various questions to the chat engine, how it can contribute to neurosurgery education by preparing case reports or questions, and its contributions when writing academic articles. The results of the study showed that while ChatGPT provided intriguing and interesting responses, it should not be considered a dependable source of information. The absence of citations for scientific queries raises doubts about the credibility of the answers provided. Therefore, it is not advisable to solely rely on ChatGPT as an educational resource. With further updates and more specific prompts, it may be possible to improve its accuracy. In conclusion, while ChatGPT has potential as an educational tool, its reliability needs to be further evaluated and improved before it can be widely adopted in neurosurgical education.

Innervation of the heart: Anatomical study with application to better understanding pathologies of the cardiac autonomics.

Current advances in management of the cardiac neuroaxis in different cardiovascular diseases require a deeper knowledge of cardiac neuroanatomy. The aim of the study was to increase knowledge of the human fetal extrinsic cardiac nervous system. We achieved this by systematizing the origin and formation of the cardiac nerves, branches, and ganglia and their sympathetic/parasympathetic connections. Thirty human fetuses (60 sides) were subjected to detailed sub-macroscopic dissection of the cervical and thoracic regions. Cardiac accessory ganglia lying on a cardiac nerve or in conjunction with two or more (up to four) nerves before entering the mediastinal cardiac plexus were observed in 13 sides. Except for the superior cardiac nerve, the sympathetic cardiac nerves were individually variable and inconstant. In contrast, the cardiac branches of the vagus nerve appeared grossly more constant and invariable, although the individual cardiac branches varied in number and position of origin. Each cervical cardiac nerve or cardiac branch of the vagus nerve could be singular or multiple (up to six) and originated from the sympathetic trunk or the vagus nerve by one, two, or three roots. Sympathetic nerves arose from the cervical-thoracic ganglia or the interganglionic segment of the sympathetic trunk. Connections were found outside the cardiac plexus. Some cardiac nerves were connected to non-cardiac nerves, while others were connected to each other. Common sympathetic/parasympathetic cardiac nerve trunks were more frequent on right (70%) versus left sides (20%). The origin, frequency, and connections of the cardiac nerves and branches are highly variable in the fetus. Detailed knowledge of the normal neuroanatomy of the heart could be useful during cardiac neuromodulation procedures and in better understanding nervous pathologies of the heart.

Human lumbar sympathetic blockade: An anatomical study to address potential block failure.

The lumbar sympathetic block is often used to treat complex regional pain syndrome, but it seems to have a high failure rate. This study seeks anatomical explanations for this apparent failure in order to refine our block procedure. Two simulated sympathetic trunk blocks were carried out on four fresh, cryopreserved unembalmed human cadavers under fluoroscopic control at the L2 vertebral body level, followed by two further simulated blocks at the L4 vertebral body level on the other side. Dye was injected, and the areas were dissected following a specific protocol. We then describe the anatomy and the spread of the dye compared to the spread of the contrast medium on fluoroscopy. The ganglia were differently located at different vertebral levels, and differed among the cadavers. Following this anatomical clarification, we now prefer to perform lumbar sympathetic blocks at the fourth lumbar vertebra level, using an extraforaminal approach at the caudal end of ​​the vertebra, avoiding the anterolateral margin of the vertebral body at the midpoint.

The lateral ulnar collateral ligament: Anatomical and structural study for clinical application in the diagnosis and treatment of elbow lateral ligament injuries.

The lateral ulnar collateral ligament (LUCL) is considered one of the main stabilizers of the elbow. However, its anatomical description is not well established. Imaging techniques do not always have agreed upon parameters for the study of this ligament. Therefore, herein, we studied the macro and microanatomy of the LUCL to establish its morphological and morphometric characteristics more precisely. Fifty-five fresh-frozen human elbows underwent dissection of the lateral collateral ligament. Morphological characteristics were studied in detail. Ultrasound (US) and magnetic resonance (MR) were done before dissection. Two specimens were selected for PGP 9.5 S immunohistochemistry. Ten additional elbows were analyzed by E12 sheet plastination. LUCL was identified in all specimens and clearly defined by E12 semi-thin sections. It fused with the common extensor tendon and the radial ligament. The total length of the LUCL was 48.50 mm at 90°, 46.76 mm at maximum flexion and 44.10 mm at complete extension. Three morphological insertion variants were identified. Both US and MR identified the LUCL in all cases. It was hypoechoic in the middle and distal third in 85%. The LUCL was hypointense on MR in 95%. Free nerve endings were present on histology. The LUCL is closely related to the anular ligament. It is stretched during flexion and supination. US and MR can reliably identify its fibers. Anatomical data are relevant to the surgeon who repairs the ligaments of the elbow. Also, to the radiologist and pain physician who interpret imaging and treat patients with pain syndromes of the elbow.

How to Differentiate Pronator Syndrome from Carpal Tunnel Syndrome: A Comprehensive Clinical Comparison.

The diagnostic process that allows pronator syndrome to be differentiated reliably from carpal tunnel syndrome remains a challenge for clinicians, as evidenced by the most common cause of pronator syndrome misdiagnosis: carpal tunnel syndrome. Pronator syndrome can be caused by compression of the median nerve as it passes through the anatomical structures of the forearm, while carpal tunnel syndrome refers to one particular topographic area within which compression occurs, the carpal tunnel. The present narrative review is a complex clinical comparison of the two syndromes with their anatomical backgrounds involving topographical relationships, morphology, clinical picture, differential diagnosis, and therapeutic options. It discusses the most frequently used diagnostic techniques and their correct interpretations. Its main goal is to provide an up-to-date picture of the current understanding of the disease processes and their etiologies, to establish an appropriate diagnosis, and introduce relevant treatment benefiting the patient.

A comparative study of the epiligament of the medial collateral and anterior cruciate ligaments in the human knee: Immunohistochemical analysis of CD 34, α-smooth muscle actin and vascular endothelial growth factor in relation to epiligament theory.

BACKGROUND: This study evaluated and compared the expression of VEGF, CD34, and α-SMA in the anterior cruciate ligaments and medial collateral ligaments in healthy human knees in order to enrich the epiligament theory regarding ligament healing after injury. METHODS: Samples from the mid-substance of the anterior cruciate ligament and the medial collateral ligament of 12 fresh knee joints were used. Monoclonal antibodies against CD34, α-SMA, and VEGF were used for immunohistochemical analysis. Photomicrographs were analyzed using the ImageJ software. RESULTS: The epiligament of the anterior cruciate ligament showed slightly higher expression of CD34, α-SMA, and VEGF than the epiligament of the medial collateral ligament. Overall, among the tested markers, α-SMA expression was most pronounced in anterior cruciate ligament epiligament images and CD34 dominated in medial collateral ligament epiligament images. The intensity of DAB staining for CD34, α-SMA, and VEGF was higher in vascular areas of the epiligament than in epiligament connective tissue. CONCLUSIONS: The results illustrate that CD34, α-SMA, and VEGF are expressed in the human epiligament. The differences between the epiligament of the investigated ligaments and the fact that CD34, α-SMA, and VEGF, which are known to have a definite role in ligament healing, are predominantly expressed in the main vascular part of the ligament-epiligament complex enlarge the existing epiligament theory. Future investigations regarding better ligament healing should not overlook the epiligament tissue.

Microscopy of structures surrounding typical acupoints used in clinical practice and electron microscopic evaluation of acupuncture needles.

Although the general functionality and structures of acupoints have been studied, there has been little insight into their underlying morphology and physical characteristics. We describe the microanatomical structures surrounding acupoints, the electron microscopic appearance of the needles, and the physical effects of acupuncture needling on the fascia. We injected heparinized blood solution through thin needles at seven known and commonly used "sweat acupoints" in eight fresh, unembalmed, cryopreserved human cadavers to mark the needle positions, and later, during histological examination, to identify them. After the solution was injected, samples were dissected and prepared for histological examination. We examined 350 cross-sections of five different paraffin wax sections from each acupoint microscopically. Acupuncture needles were photographed and superimposed on the cross-sectioned tissues at similar magnifications. Needles were also examined under a scanning electron microscope to judge the roughness or smoothness of their surfaces. A greater conglomeration of nerve endings surrounded the acupoints than in tissues more than 1-3 cm distant from them. Nerve endings and blood vessels were in close contact with a complex network of membranes formed by interlacing collagen fibers, and were always enclosed within those collagen membranes. Nerve endings were found within hypodermis, muscles, or both. Scanning electron microscopy demonstrated the three-dimensional shapes and sizes of the needles, and the degree of roughness or smoothness of their polished external surfaces. We demonstrate a delicate arrangement of nerve endings and blood vessels enclosed within complex collagen membrane networks at acupoints within the hypodermis and muscle. This arrangement could explain why needling is an essential step in the acupuncture process that provides favorable outcomes in clinical practice.