PURPOSE: To investigate the regulatory role of nerve injury-induced protein 1 (NINJ1) in the anti-inflammatory function of human umbilical cord mesenchymal stem cells (hUC-MSCs) co-stimulated by interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α). METHODS: hUC-MSCs were expanded in vitro using standard protocols, with stem cell characteristics confirmed by flow cytometry and multilineage differentiation assays. The immunomodulatory properties and cellular activity of cytokine-co-pretreated hUC-MSCs were systematically evaluated via quantitative reverse transcription RT-qPCR, lymphocyte proliferation suppression assays, and Cell Counting Kit-8 viability tests. Transcriptome sequencing, Western blotting and small interfering RNA interference were integrated to analyze the regulatory mechanisms of NINJ1 expression. Functional roles of NINJ1 in pretreated hUC-MSCs were elucidated through gene silencing combined with lactate dehydrogenase release assays, Annexin V/Propidium Iodide apoptosis analysis, macrophage co-culture models, and cytokine Enzyme-Linked Immunosorbent Assay. Therapeutic efficacy was validated in a cecal ligation and puncture-induced septic mouse model: 80 mice were randomly allocated into 4 experimental groups (n=20/group): sham group (laparotomy without cecal ligation); phosphate-buffered saline-treated group (cecal ligation and puncture (CLP) + 0.1 mL phosphate-buffered saline); hUC-MSCs (small interfering RNA (siRNA)-interferon-gamma and tumor necrosis factor-alpha co-stimulation (IT))-treated group (CLP + hUC-MSCs transfected with scrambled siRNA); and hUC-MSCs (siNINJ1-IT)-treated group (CLP + hUC-MSCs with NINJ1-targeting siRNA). RESULTS: hUC-MSCs demonstrated compliance with International Society for Cellular Therapy criteria, confirming their stem cell identity. IFN-γ/TNF-α co-pretreatment enhanced the immunosuppressive capacity of hUC-MSCs, accompanied by the reduction of cellular viability, while concurrently upregulating pro-inflammatory cytokines such as interleukin-6 and interleukin-1β. This co-stimulation significantly elevated NINJ1 expression in hUC-MSCs, whereas genetic silencing of NINJ1 effectively suppressed pro-inflammatory cytokine production and attenuated damage-associated molecular patterns release through inhibition of programmed plasma membrane rupture. Furthermore, the NINJ1 interference potentiated the ability of cytokine-pretreated hUC-MSCs to suppress LPS-induced pro-inflammatory responses in RAW264.7 macrophages. In cecal ligation and puncture-induced sepsis model, NINJ1-silenced hUC-MSCs exhibited enhanced therapeutic efficacy, manifested by reduced systemic inflammation and multi-organ damage. CONCLUSION Our findings shed new light on the immunomodulatory functions of cytokine-primed MSCs, offering groundbreaking insights for developing MSC-based therapies against inflammatory diseases via interfering the expression of NINJ1.
Mesenchymal Stem Cells/drug effects* ; Animals ; Interferon-gamma/pharmacology* ; Tumor Necrosis Factor-alpha/pharmacology* ; Humans ; Mice ; Umbilical Cord/cytology* ; Cells, Cultured ; Apoptosis ; Male

Anxiety/epidemiology* ; Depression/epidemiology* ; Humans ; Sleep ; Sleep Quality ; Students ; Surveys and Questionnaires ; Universities

The present study evaluates different processing and drying methods and investigates their effects on the chemical components in Paeoniae Radix Alba via content determination. The fresh medicinal materials of Paeoniae Radix Alba collected from Bozhou of Anhui province were processed(boiled and peeled) and dried(hot air-dried, infrared-dried, and microwave-dried) at different temperatures(40, 50, 60 and 70 ℃), and the 11 components(monoterpene glycosides, polyphenols, tannin, and benzoic acid) in Paeoniae Radix Alba were determined by ultra-performance liquid chromatography coupled to triple quadrupole with electrospray tandem mass spectrometry(UPLC-TQ-MS). Then the compounds in processed and dried samples were analyzed by partial least squares discriminant analysis(PLS-DA) and orthogonal partial least squares discriminant analysis(OPLS-DA), and the contribution rates of differential components were evaluated by variable important in projection(VIP). The results indicated that the samples obtained by different processing and drying methods could be distinguished. Albiflorin, gallic acid, 1,2,3,4,6-pentakis-O-galloyl-β-D-glucose, and benzoic acid were the common differential components in boiled Paeoniae Radix Alba. Benzoic acid was the common differential component in peeled Paeoniae Radix Alba. Gallic acid was the common differential component in Paeoniae Radix Alba dried by different methods. The samples could not be distinguished after drying at different temperatures due to the lack of common differential components. This study is expected to provide a reference for the selection of processing and drying methods and the optimization of processing parameters.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Paeonia ; Plant Extracts ; Tandem Mass Spectrometry

OBJECTIVE: To observe the changes of functional connectivity of brain pain-emotion regulation region in patients with cervical spondylosis of cervical type by functional magnetic resonance imaging (fMRI). METHODS: Thirty-two subjects were selected. Of them, 16 patients with cervical spondylosis of cervical type were divided into an observation group and 16 healthy subjects into a control group. The patients in the observation group were treated with acupuncture at Tianzhu (BL 10), Jingbailao (EX-HN 15), Jianzhongshu (SI 15) and RESULTS: In the observation group, the VAS score was (1.94±1.12) after the treatment, which was lower than (5.62±1.20) before treatment ( CONCLUSION Pain involves the formation and expression of "pain-emotion-cognition". Acupuncture can systematically regulate the brain functional connections between cognitive regions such as dorsal prefrontal lobe and anterior cingulate gyrus and emotional regions such as insula and VTA in patients with cervical spondylosis of cervical type, suggesting that acupuncture has a multi-dimensional and comprehensive regulation effect on pain.
Acupuncture Therapy ; Brain/diagnostic imaging* ; Emotions ; Humans ; Magnetic Resonance Imaging ; Pain ; Spondylosis/therapy*

This study was designed to determine the metabolites of Zhali Nusi Prescription(ZLNSP) in rats. The ultra-high performance liquid chromatography-LTQ Orbitrap mass spectrometric(UHPLC-LTQ-Orbitrap-MS) and mass defect filter techniques were applied to analyze the metabolites of ZLNSP in rat plasma, bile, urine and feces. The biological samples were analyzed by ACQUITY UPLC BEH T_3 column(2.1 mm×100 mm,1.7 μm), with 0.1% formic acid water(A)-acetonitrile(B) as mobile phase, and the biological samples were analyzed in negative ion mode by electrospray ionization mass spectrometry(ESI-MS). An analytical method for biological samples of rats was established, and 8 prototype components and 36 metabolites were identified. The results showed that the metabolic pathways of the main components of ZLNSP in rats included methylation, glucuronidation, sulfation and so on. It provi-ded information for the therapeutic effect of ZLNSP in vivo.
Administration, Oral ; Animals ; Bile ; Chromatography, High Pressure Liquid ; Feces ; Plasma ; Prescriptions ; Rats

With an objective to provide an experimental basis for scientific officinal of Schisandrae Sphenantherae Fructus, this research uses UPLC-TQ/MS method to analyze 7 different kinds of lignan in 70 batches of Schisandra sphenantherae Fructus samples from 9 regions. The results showed that in the area south of Qinling mountains, Schisandrae sphenantherae Fructus from Zhashui county and Shanyang county of Shangluo mainly contained schisantherin A and deoxyschizandrin. However, Schisandrae sphenantherae Fructus from Mei county of Baoji, Shiquan county and Ningshan county of Ankang, and Lueyang county of Hanzhong, mainly contained anwuligan. Samples from Ningshan county also consists relatively high level of deoxyschizandrin. In the central area of Qinling mountains and the Daba mountains, Schisandrae Sphenantherae Fructus from Nanzheng county of Hanzhong mainly contained schisanhenol and deoxyschizandrin. In conclusion, the kinds and level of lignan differ significantly in Schisandrae sphenantherae Fructus produced in different regions. In practical application, Schisandrae Sphenantherae Fructus produced in different regions should be distinguished and differently applied based on their main effective components corresponding to different diseases, which can lead to the best clinical use.
China ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Lignans ; chemistry ; Quality Control ; Schisandra ; chemistry

BACKGROUNDProprioception plays an important role in knee movements. Since there are controversies surrounding the overall recovery time of proprioception following surgery, it is necessary to define the factors affecting proprioceptive recovery after anterior cruciate ligament (ACL) reconstruction and to investigate the relationship between proprioception and muscle strength.

METHODSA total of 36 patients who had their ACL reconstructed with a semitendinosus/gracilis graft (reconstructed group: 6 months post-surgery) and 13 healthy adults without any knee injury (control group) were included in the study. Knee proprioception was evaluated with a passive reproduction test. Isokinetic strength was measured using the Biodex System. Statistical analysis was used to compare proprioception of the reconstructed group versus the control group, and to define causal factors, including sex, hamstring/quadriceps ratio, and the course of injury before reconstruction. We also investigated the correlation between the passive reproduction error and quadriceps index.

RESULTSThere was a significant difference in proprioception between the reconstructed and control groups (P < 0.05). When the course of injury before reconstruction was less than 4 months, there was a linear correlation with proprioception 6 months after the operation (r = 0.713, P < 0.05). There was a positive correlation between post-surgery proprioception and the quadriceps index at 6 months post-surgery.

CONCLUSIONSImpaired knee proprioception is observed 6 months after ACL reconstruction. Within 4 months of injury, early undertaking of reconstruction is associated with better proprioception outcome. Patients with enhanced proprioception have a better quadriceps index.

Adolescent ; Adult ; Anterior Cruciate Ligament ; pathology ; physiopathology ; surgery ; Female ; Humans ; Knee Injuries ; pathology ; physiopathology ; surgery ; Male ; Proprioception ; physiology ; Reconstructive Surgical Procedures ; methods ; Recovery of Function ; Young Adult

BACKGROUNDAfter anterior cruciate ligament (ACL) reconstruction, some patients suffered from sensory disturbance around the surgical incision of the leg. This research was aimed to investigate the relationship between the different skin incisions and the injury of the infrapatellar branch of the saphenous nerve (IPBSN) post ACL reconstruction.

METHODSACL reconstructions were performed with quadruple hamstring tendon for 60 patients. Sensory disturbance around the skin incision was followed up at an average of 14.5 +/- 4.7 months post operation. Among the 60 patients, vertical incision for 35 patients and oblique incision for 25 patients were used for graft taking during ACL reconstruction. The lengths of the incisions were measured. The patients were asked to mark the sensory disturbance zone at follow up time, and then the marked area was measured. The IPBSN of 15 cadaver knees were anatomized. The distance between the IPBSN and the upper edge of the pes anserinus tendon at the middle point of the incision was measured. Independent-samples t-test, chi-square and Mann-Whitney tests were used for statistical analyses.

RESULTSThe patients' age (P = 0.329), the follow-up time (P = 0.681), and the incision length (P = 0.732) between the two groups had no significant difference. Twenty-three patients (65.7%) in the vertical incision group had IPBSN injury compared with 6 patients (24.0%) in oblique incision group (P = 0.002). The average sensory disturbance area in vertical incision group ((48.0 +/- 75.3) cm(2)) was significantly larger (P = 0.004) than that in the oblique group ((8.4 +/- 19.4) cm(2)). The anatomy measurement showed the average distance between IPBSN and the upper edge of the pes anserinus tendon was 0.6 cm at the incision.

CONCLUSIONSOblique incision with less risk of damage for IPBSN may be better for graft harvesting in ACL reconstruction. As the IPBSN is so near and parallel to the hamstring tendons, damage to the IPBSN is one of the potential complications for graft harvesting, regardless of the incision used. That's why even in the oblique incision group, 24% patients also had sensory disturbance complication.

Adolescent ; Adult ; Anterior Cruciate Ligament ; surgery ; Female ; Humans ; Knee Joint ; innervation ; Male ; Middle Aged ; Peripheral Nerve Injuries ; Reconstructive Surgical Procedures ; adverse effects