Objective To investigate the potential therapeutic efficacy of lentivirus-mediated artemin (ARTN) gene modified bone marrow mesenchymal stem cells (MSCs) transplantation on the rat model of Parkinson' s disease (PD) and the effects on expression of brain-related proteins.Methods MSCs were isolated and cultured in vitro,transfected by recombinant lentiviral vectors carrying ARTN gene.The PD rat model established by 6-hydroxydopamine (6-OHDA) was randomly divided into 5 groups:Sham group,PD group,MSCs group,MSCs transfected with empty lentiviral vectors transplanted (LV-MSCs)group and MSCs transfected with recombinant lentiviral vectors carrying ARTN gene transplanted (LVARTN-MSCs) group.The MSCs,LV-MSCs and LV-ARTN-MSCs groups were transplanted into the left striatum of each rat model of PD and ethology tests in every group were made with intraperitoneal injection of apomorphine (APO) 2,4,6,8 weeks after transplantation.The expression of tyrosine hydroxylase (TH) protein in substantia nigra (SN) was measured by Western blotting and immunohistochemistry,and immunofluorescence showed ARTN gene modified MSCs expression in rat brain tissue.The levels of dopamine (DA),dihydroxy-phenylacetic acid and homovanillic acid in striatum of each group were detected by high performance liquid chromatography.Results After injection of APO,rotation frequency decreased in LV-ARTN-MSCs group,i.e.(179.33 ± 10.74) circles/30 min vs (235.83 ± 18.95),(203.67 ±11.50) and (206.33 ± 11.86) circles/30 min in PD,MSCs and LV-MSCs groups (q =8.828,P ＜ 0.01;q =3.802,P ＜ 0.05;q =4.219,P ＜ 0.05).The percentage of TH-positive cells in SN after cell transplantation was increased significantly in LV-ARTN-MSCs group (64.05％ ± 5.49％) when compared with PD group (34.18％ ±3.35％),MSCs group (52.59％ ±4.48％) and LV-MSCs group (50.57％ ± 4.41％),respectively (q =13.280,5.135,6.028,all P ＜0.01).At the same time,TH protein in SN after cell transplantation was also increased obviously in LV-ARTN-MSCs group.ARTN gene modified MSCs can survive for at least 6 weeks in the rat brain of PD,mainly concentrated in the transplantation side of striatum.Eight weeks later,the levels of DA in striatum after cell transplantation were elevated significantly in MSCs group (2.34 ± 0.54),LV-MSCs group (2.28 ± 0.45) and LV-ARTN-MSCs group (2.28 ± 0.45)when compared with PD group (0.87 ± 0.07) (q =5.233,P ＜ 0.05;q =5.020,P ＜ 0.01;q =20.190,P ＜ 0.01),and LV-ARTN-MSCs group showed the most significant improvement.Conclusion ARTN gene modified bone marrow MSCs transplanted into the striatum of brain may have therapeutic effects on rat models of PD.

Objective:To analyze the magnetic resonance imaging (MRI) features of diffuse midline gliomas with H3K27M mutation, and to quantitatively analyze the changes of apparent diffusion coefficient (ADC).Methods:The MRI images of 14 cases of diffuse midline gliomas with H3K27M mutation were retrospectively analyzed in the Affiliated Hospital of Qingdao University from April 2017 to November 2019. The location, edge, signal, peritumoral edema and enhancement characteristics of the lesions were observed, and the changes of ADC values were analyzed.Results:The tumors were located in thalamus in four cases, pons in six cases, medulla oblongata in two cases and spinal cord in two cases. In seven cases, the tumor was confined to the midline region, of which six cases had clear boundary, seven cases were located in the midline area, but infiltrated into the non midline area at the same time, and six cases had unclear boundary. Basilar artery entrapment was found in all six patients located in pons. Multiple large cystic changes were found in five cases, multiple small cysts in four cases and no cystic changes in five cases. Cystic changes were found in all seven cases of tumors involving the non midline region, of which six cases were located only in the non midline region, and only two of the seven tumors localized in the midline region had small cysts. Hemorrhage was found in four cases. Five cases showed mild heterogeneous enhancement, six cases showed moderate heterogeneous enhancement, two cases showed obvious enhancement, and one case showed no enhancement. There was no edema around the tumor in nine cases and mild edema in five cases. The average edema index was 1.13. The average ADC value of tumor parenchyma in 12 patients was (7.83±0.88)×10 -4 mm 2/s, which was 15.6% lower than that of the contralateral side [(9.28±0.69)×10 -4 mm 2/s, t=-6.336, P<0.05]. Conclusions:Diffuse midline gliomas with H3K27M mutation have a younger onset age and are more likely to occur in thalamus, brainstem and spinal cord. Most of the tumors have no peritumoral edema or mild peritumoral edema. The tumors confined to the midline region are regular in shape and clear in boundary. The masses involving the non midline area are prone to cystic necrosis. Diffuse midline gliomas with H3K27M mutation in pons are prone to basilar artery entrapment. ADC value can provide a quantitative basis for preoperative tumor grading.

Objective:To compare the short-term and long-term efficacies and safety of short-term spinal cord stimulation (st-SCS) and pulsed radiofrequency (PRF) in treating zoster-associated pain (ZAP).Methods:A total of 208 ZAP patients who received treatment in Department of Pain Management, Provincial Hospital Affiliated to Shandong University from January 2019 to September 2021 were selected. They were divided into st-SCS group ( n=107) and PRF group ( n=101) according to different treatments. After differences of baseline data being balanced by 1∶1 propensity score matching between the two groups, scores of numerical rating scale (NRS), NRS remission rate, drug dosage, Pittsburgh Sleep Quality Index (PSQI) and complications of 86 patients from each 2 groups were compared and analyzed before surgery, 3 and 7 d after surgery, at discharge, and 1, 3, 6, 12 and 24 months after surgery. Results:NRS scores in st-SCS group and PRF group 3 and 7 d after surgery, at discharge, and 1, 3, 6, 12 and 24 months after surgery were significantly lower than those before surgery ( P<0.05). NRS scores in st-SCS group were significantly lower than those in PRF group at discharge and 1, 3, 6 and 12 months after surgery ( P<0.05). NRS remission rate in st-SCS group was significantly higher than that in PRF group at discharge and 1, 3, 6, 12 and 24 months after surgery, and total effective rate in st-SCS group was significantly higher than that in PRF group 1, 3, 6, and 12 months after surgery ( P<0.05). The pregabalin and gabapentin dosages in st-SCS group and PRF group 1, 3, 6, 12, and 24 months after surgery were significantly lower than those before surgery, and the gabapentin dosage in st-SCS group was significantly lower than that in PRF group 3 months after surgery ( P<0.05). PSQI in st-SCS group and PRF group at discharge and 6, 12 and 24 months after surgery was significantly decreased compared with that before surgery, and PSQI in st-SCS group at discharge and 6, 12 and 24 months after surgery was significantly decreased compared with that in PRF group ( P<0.05). No serious complications as spinal epidural hematoma or spinal nerve root and spinal cord injuries occurred in st-SCS group and PRF group. Conclusion:Both st-SCS and PRF can treat ZAP safely and effectively, but st-SCS has more advantages than PRF.

Objective To investigate the value of intravoxel incoherent motion DWI (IVIM-DWI) in differential diagnosis of high-grade gliomas and brain metastases.Methods Conventional MRI,contrast-enhanced MRI and IVIM-DWI were performed before surgery or chemoradiotherapy in 24 patients with high-grade gliomas and 28 patients with brain metastases.The diffusion constant (D),pseudodiffusion coefficient of perfusion (D*) and the perfusion fraction (f) in the parenchyma and peritumoral edema region within 1 cm and the normal centrum semiovale in the opposite side were measured,then the relative values of all parameters in each region (rD*,rD,rf) were calculated.Independent sample t test was used to analyze the parameters.ROC curve analysis of the parameters statistically different between high grade gliomas and brain metastases were performed,and the diagnostic efficacies were evaluated.Results The D* and rD* values of tumor parenchyma and in peritumoral edema within 1 cm of high-grade gliomas were higher than those of brain metastases (all P＜0.05).The f and rf values of tumor parenchyma and in peritumoral edema within 1 cm of highgrade gliomas were lower than those of brain metastases (all P＜0.01).The AUC of D* value in peritumoral edema within 1 cm was the highest,but there was no statistically different between any two AUC except the rD* value of peritumoral edema within 1 cm (P =0.033).Conclusion IVIM-DWI can distinguish the differences of diffusion and perfusion information in parenchyma and edema area between high-grade gliomas and brain metastases,therefore providing the basis for differential diagnosis of them.

Objective To discuss the protective effect of Artemin (ARTN) on 6-hydroxydopamine (6-OHDA)-mediated neurotoxic injury,and the alternations of related proteins in Parkinson's disease (PD) model rats.Methods Bone marrow mesenchymal stem cells (MSCs) were isolated and cultured in vitro.After being transfected with recombinant lentiviral vectors carrying A RTN gene,MSCs stably expressed ARTN were chosen (Lv-ARTN-MSCs cells).The SH-SY5Y cells were treated with supematant of Lv-ARTN-MSCs prior to 6-OHDA treatment,and then,cell survival rate was measured by MTT assay and morphologic changes in cultured SH-SY5Y cells were observed by fluorescence microscope (Hochest33258 staining).PD rat models were established and randomly divided into four groups (n=6):PD group,MSCs group,Lv-MSCs group and Lv-ARTN-MSCs group; and sham-operated group (n=6) was also chosen.The PD,MSCs,Lv-MSCs and Lv-ARTN-MSCs groups were transplanted with 5 μL of saline,MSCs (1.0×l05 cell/5 μL),empty virus modified MSCs (1.0×105 cell/5 μL) and ARTN gene modified MSCs (1.0×105 cell/5 μL),respectively,into the left striatum; rats in the sham-operated group were injected with saline when rats in the other groups were received 6-OHDA injection (the same surgical procedures and coordinates).The expressions of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the striatum were measured by Western blotting.Results Western blotting indicated obvious ARTN protein expressions in the Lv-ARTN-MSCs groups.The supematant of Lv-ARTN-MSCs could effectively reduce the apoptosis rate induced by 6-OHDA; as compared with that of SH-SY5Y cells in the 6-OHDA group,the cell survival rate in the Lv-A R TN-MSCs group increased by 13.67％,with significant difference (P＜0.05).Eight weeks after transplantation,the levels of TH and DAT protein in the striatum were elevated significantly in MSCs group,Lv-MSCs group and Lv-ARTN-MSCs group as compared with those in the PD group (P＜0.05),and the Lv-ARTN-MSCs group showed the most significant improvement.Conclusion ARTN which is a secreted protein can protect dopaminergic neuron against 6-OHDA-induced toxicities in Parkinson's disease,and the mechanism might be related to the increased expressions of TH and DAT.

Objective:To investigate the effect of vector sum concept in fine-tuning posterior column screw channel via ilioinguinal approach for the treatment of bi-column acetabular fracture.Methods:A retrospective cohort study was conducted to analyze the clinical data of 42 patients with acetabular double column fracture admitted to Weifang People′s Hospital from July 2015 to May 2021, including 22 males and 20 females, aged 23-77 years [(49.3±16.3)years]. The ilioinguinal approach was used in all patients with the anterior column fixed with a plate and posterior column fixed with a lag screw. The vector sum concept was used intraoperatively to fine-tune the posterior column screw channel in 19 patients (channel fine-tuning group): namely, a 2.5 mm Kirschner wire was inserted into the bony channel of the posterior column screw under fluoroscopy of iliac oblique and obturator oblique positions; when the Kirschner wire was not located in the middle of the ischial ramus under single fluoroscopy, the vector only needed to be adjusted in one direction, with zero in the other direction; when the Kirschner wire was not located in the middle of the ischial ramus under fluoroscopy of both the iliac oblique and obturator obturator oblique positions, the sum of the deviation vectors in the two directions was calculated before fine-tuning. The vector sum concept was not used to fine-tune the posterior column channel screw in 23 patients (channel non-fine-tuning group). The time of posterior column screw placement, intraoperative blood loss, frequency of guide wire adjustment and fracture healing time were recorded and compared between the two groups. At 6 months after operation, the quality of fracture reduction and hip function were assessed by Matta score and Merle D′Aubigne-Postel score, respectively. The complications were observed.Results:All patients were followed up for 7-71 months [(35.7±8.5)months]. In channel fine-tuning group, the time of posterior column screw placement was (5.1±1.5)minutes, with intraoperative blood loss of (798.8±83.9)ml, frequency of guide wire adjustment of (1.8±0.5)times and fracture healing time of (12.4±3.2)weeks; while these parameters [(39.8±12.0)minutes, (1 119.3±172.0)ml, (5.6±1.6)times and (15.6±4.2)weeks] were significantly shorter or less in channel non-fine-tuning group ( P<0.05 or 0.01). There were no significant difference in the quality of fracture reduction and hip function between the two groups at 6 months postoperatively (all P>0.05). After operation, symptoms of lateral femoral cutaneous nerve was found in seven patients, superficial incision infection in two who was healed after debridement and dressing change, deep venous thrombosis of lower limbs in three. There was no significant difference in the incidence of postoperative complications between the two groups [channel fine-tuning group: 26%(5/19), channel non-fine-tuning group: 30%(7/23)] ( P>0.05). Conclusion:For bi-column acetabular fractures via ilioinguinal approach, application of vector and concept to fine-tune the posterior column screw channel is beneficial for rapid screw placement into the osseous channel, significant reduction of intraoperative blood loss and early fracture healing.

Tolerogenic dendritic cells (tolDCs) facilitate the suppression of autoimmune responses by differentiating regulatory T cells (Treg). The dysfunction of immunotolerance results in the development of autoimmune diseases, such as rheumatoid arthritis (RA). As multipotent progenitor cells, mesenchymal stem cells (MSCs), can regulate dendritic cells (DCs) to restore their immunosuppressive function and prevent disease development. However, the underlying mechanisms of MSCs in regulating DCs still need to be better defined. Simultaneously, the delivery system for MSCs also influences their function. Herein, MSCs are encapsulated in alginate hydrogel to improve cell survival and retention in situ, maximizing efficacy in vivo. The three-dimensional co-culture of encapsulated MSCs with DCs demonstrates that MSCs can inhibit the maturation of DCs and the secretion of pro-inflammatory cytokines. In the collagen-induced arthritis (CIA) mice model, alginate hydrogel encapsulated MSCs induce a significantly higher expression of CD39⁺CD73⁺ on MSCs. These enzymes hydrolyze ATP to adenosine and activate A_2A/2B receptors on immature DCs, further promoting the phenotypic transformation of DCs to tolDCs and regulating naïve T cells to Tregs. Therefore, encapsulated MSCs obviously alleviate the inflammatory response and prevent CIA progression. This finding clarifies the mechanism of MSCs-DCs crosstalk in eliciting the immunosuppression effect and provides insights into hydrogel-promoted stem cell therapy for autoimmune diseases.

Cystic fibrosis (CF) is a rare autosomal recessive disease with only one pathogenic gene cystic fibrosis transmembrane conductance regulator (CFTR). To identify the potential pathogenic mutations in a Chinese patient with CF, we conducted Sanger sequencing on the genomic DNA of the patient and his parents and detected all 27 coding exons of CFTR and their flanking intronic regions. The patient is a compound heterozygote of c.2909G > A, p.Gly970Asp in exon 18 and c.1210-3C > G in cis with a poly-T of 5T (T5) sequence, 3 bp upstream in intron 9. The splicing effect of c.1210-3C > G was verified via minigene assay in vitro, indicating that wild-type plasmid containing c.1210-3C together with T7 sequence produced a normal transcript and partial exon 10-skipping-transcript, whereas mutant plasmid containing c.1210-3G in cis with T5 sequence caused almost all mRNA to skip exon 10. Overall, c.1210-3C > G, the newly identified pathogenic mutation in our patient, in combination with T5 sequence in cis, affects the CFTR gene splicing and produces nearly no normal transcript in vitro. Moreover, this patient carries a p.Gly970Asp mutation, thus confirming the high-frequency of this mutation in Chinese patients with CF.
China ; Cystic Fibrosis/genetics* ; Cystic Fibrosis Transmembrane Conductance Regulator/genetics* ; Humans ; Mutation ; Poly T ; RNA, Messenger/genetics*

Primary ciliary dyskinesia (PCD) is a highly heterogeneous recessive inherited disorder. FAP54, the homolog of CFAP54 in Chlamydomonas reinhardtii, was previously demonstrated as the C1d projection of the central microtubule apparatus of flagella. A Cfap54 knockout mouse model was then reported to have PCD-relevant phenotypes. Through whole-exome sequencing, compound heterozygous variants c.2649_2657delinC (p. E883Dfs*47) and c.7312_7313insCGCAGGCTGAATTCTTGG (p. T2438delinsTQAEFLA) in a new suspected PCD-relevant gene, CFAP54, were identified in an individual with PCD. Two missense variants, c.4112A>C (p. E1371A) and c.6559C>T (p. P2187S), in CFAP54 were detected in another unrelated patient. In this study, a minigene assay was conducted on the frameshift mutation showing a reduction in mRNA expression. In addition, a CFAP54 in-frame variant knock-in mouse model was established, which recapitulated the typical symptoms of PCD, including hydrocephalus, infertility, and mucus accumulation in nasal sinuses. Correspondingly, two missense variants were deleterious, with a dramatic reduction in mRNA abundance from bronchial tissue and sperm. The identification of PCD-causing variants of CFAP54 in two unrelated patients with PCD for the first time provides strong supportive evidence that CFAP54 is a new PCD-causing gene. This study further helps expand the disease-associated gene spectrum and improve genetic testing for PCD diagnosis in the future.
Mice ; Animals ; Humans ; Male ; Kartagener Syndrome/metabolism* ; Cilia/metabolism* ; Semen ; Genetic Testing ; RNA, Messenger ; Mutation