AIM: To investigate the effect of intense pulsed light(IPL)combined with meibomian gland massage in patients with meibomian gland dysfunction(MGD)related dry eye.METHODS: Retrospectively selected 300 cases(300 right eyes)that diagnosed as MGD-related dry eye and treated in our hospital from October 2021 to October 2023, and they were divided into two groups according to the treatment modalities: 150 cases(150 eyes)in the control group were treated with meibomian gland massage, and 150 cases(150 eyes)in the observation group were treated with combined IPL. The efficacy of the two groups was compared, as well as the changes in tear film stability indexes [tear film break-up time(BUT), Schirmer I test(SIT)], ocular surface disease index(OSDI)scores, tear film lipid layer thickness(LLT), mean objective scattering index(OSI), and the levels of inflammatory factors in the tear fluid [interleukin-10(IL-10)and IL-6].RESULTS: There was a difference in efficacy between the two groups of patients after treatment(88% vs 74%, P=0.002). At 8 wk after treatment, OSDI score, OSI, and tear IL-6 levels decreased, while BUT, SIt, LLT, and tear IL-10 levels increased in the two groups, and the observation group was better than the control group(all P<0.05).CONCLUSION: IPL combined with meibomian gland massage for the treatment of dry eye patients with MGD is therapeutically effective, improving corneal stability and LLT, and reducing levels of OSI and inflammatory factors.

AIM To identify the in vivo metabolites of Cynanchum auriculatum Royle ex Wight extract in functional dyspepsia rats by UHPLC Q-Exactive Plus Orbitrap HRMS.METHODS The rat models for functional dyspepsia were established.The analysis was performed on a 40℃ thermostatic Hypersil GOLD C18 column(2.1 mm×100 mm,1.9 μm),with the mobile phase comprising of water(containing 0.1％formic acid)-acetonitrile(containing 0.1％formic acid)flowing at 0.3 mL/min in a gradient elution manner,and electrospray ionization source was adopted in positive and negative ion scanning.RESULTS Total 4 prototypes(baishouwubenzophenone,deacylmetaplexigenin,qingyangshengenin,syringic)and 110 metabolites were identified,12 of which were common metabolites in feces and urine,56 of which were unique metabolites in urine,42 of which were unique metabolites in feces.The metabolic pathway of prototypes contained phase Ⅰ metabolism(reduction,oxidization,etc.),phase Ⅱ metabolism(sulfonation,glucuronidation,etc.)and phase Ⅰ,Ⅱ composite reactions.CONCLUSION This effective and comprehensive method can lay the theoretical foundation for further discovery of potential active metabolites in C.auriculatum.

Objective:To investigate effect of Mycobacterium tuberculosis BCG-infected macrophages on damage and repair of renal tubular epithelial cells during development of renal tuberculosis.Methods:A co-culture model of BCG-infected M0 macrophages(upper chamber)and HK-2 cells(lower chamber)was established by Transwell,and THP-1 human monocyte macrophages were induced by 100 ng/ml phorbol ester(PMA)24 h to become M0 macrophages,and BCG infection cell model was established.Total cell protein was collected at 12 h and 24 h of infection,respectively.Western blot was used to detect expressions of M1 macrophage marker CD86 and M2 macrophage marker CD206 protein.M1 macrophage polarization marker cytokines IL-6 and TNF-α and M2 macrophage polarization marker cytokine TGF-β expressions in cell culture supernatant were detected by ELISA;experiment was divided into HK-2 group,BCG+HK-2 group,BCG+M0+HK-2 group and M0+HK-2 group,CCK-8 was used to detect viability of HK-2 cells in each group,and Hoechst test was used to detect HK-2 cells apoptosis in each group.Epithelial cell marker E-cadhren and fibroblast markerα-SMA expressions in HK-2 cells of each group were detected by Western blot.Results:After BCG infection of M0 macrophages,M1 macrophage viability was higher than 24 h at 12 h(P<0.05),and M2 macrophage was higher than 12 h at 24 h(P<0.05).After two cells co-culture,HK-2 cell viability was higher than 12 h at 24 h(P<0.001),apoptosis level was higher than 24 h at 12 h,epithelial cell marker protein E-cadherin protein level was higher than 12 h at 24 h(P<0.001),fibroblast level of cell marker protein α-SMA protein at 12 h was higher than that at 24 h(P<0.01).Conclusion:During development of renal tuberculosis,early BCG-infected macrophages may promote inflammatory injury of renal tubular epithelial cells through M1-type polarization;with prolongation of infec-tion time,they may repair renal tubular epithelial cells through M2-type polarization and plays an important protective role.

OBJECTIVE: To summarize and discuss the clinical efficacy and application value of intravenous drip of linezolid combined with local targeted sustained-release of vancomycin in the treatment of traumatic osteomyelitis of extremities infected with MRSA. METHODS: Thirty patients with traumatic osteomyelitis of extremities infected by MRSA from March 2015 to March 2017 were analyzed retrospectively, including 21 males and 9 females; aged 25 to 64 years old, with an average age of(47.94± 6.23) years old;the course of disease ranged from 9 to 23 months, with an average of (15.68±6.23) months. The lesions were located in tibia in 18 cases and calcaneus in 12 cases. The causes of injury were fall injury in 12 cases, trafficaccident injury in 9 cases and fall injury in 9 cases. There were 22 patients with closed fractures and 8 patients with open fractures. There were 13 cases of internal fixation. Twenty-two patients had sinustract, 8 patients had soft tissue defect with bone and internal fixation exposure, soft tissue defect area ranged from 2.0 cm × 3.0 cm to 8.2 cm × 12.3 cm;10 patients had bone defect, defect area ranged from 0.5 to 3.4 cm;bacterial culture of sinus tract or wound secretion in all patients was MRSA. On the basis of thorough debridement, calcium sulfate artificial bone loaded with vancomycin was implanted in the lesion, and linezolid and glucose injection was given intravenously during the perioperative period. The patients were followed up regularly according to the time of antibiotic use, blood routine, erythrocyte sedimentation rate, high-sensitivity C-reactive protein, liver and kidney function and other related laboratory indexes, X-ray, CT and other imaging examinations, bone healing, flap survival, joint function and McKee's osteomyelitis cure criteria. RESULTS: All the patients were followed up, and the duration ranged from 3 to 6 years, with a mean of (4.23±0.76) years. No recurrence of osteomyelitis occurred. Fracture healing, infection control, wound healing and functional recovery were achieved. CONCLUSION Intravenous drip of linezolid combined with local targeted sustained-release of vancomycin for the treatment of MRSA infected traumatic osteomyelitis in limbs have significant effects and low recurrence rates.
Adult ; Extremities ; Female ; Humans ; Infant ; Male ; Methicillin-Resistant Staphylococcus aureus ; Middle Aged ; Osteomyelitis ; Retrospective Studies ; Surgical Flaps ; Treatment Outcome

Objective:To investigate the changes of connexin 43 (Cx43) in human umbilical vein endothelial cells (HUVEC) after X-ray irradiation and its influence on the stiffness of irradiated cells.Methods:Western blot was used to detect the expression of Cx43 in HUVEC cells at different time points (0, 6, 12, 24 and 48 h) after different doses of X-ray irradiation (0, 2.5, 5, 10 and 20 Gy), and the phosphorylation levels of three phosphorylation sites (Ser279/282, Ser368 and Tyr265) of Cx43 at different time points (3, 6, 24 and 48 h) after 0, 5 and 10 Gy irradiation. The distribution of Cx43 protein in the irradiated HUVEC cells was detected by immunofluorescence. The stiffness changes of cells were detected by atomic force microscopy (AFM) at the depths of 50, 100 and 200 nm.Results:The expression of Cx43 in HUVEC cells was reduced at 6, 12, 24 and 48 h after 10 Gy X-ray irradiation( t=3.262, 3.708, 3.686, 6.825, P<0.05)and this decrease had a dose dependent manner at 24 h after 2.5, 5, 10 and 20 Gy irradiation ( t=3.034, 10.720, 13.130, 13.650, P<0.05). At 24 h after 5, 10 and 20 Gy X-ray irradiation, the distribution of Cx43 in HUVEC cells was transported from intercellular gap junctions to nucleus and perinuclear region. At 24-48 h after irradiation, the phosphorylation level of Ser368 at Cx43 increased and in a dose dependent manner. At 24 h after irradiation, the stiffness of the irradiated cells decreased significantly under the conditions of 100 and 200 nm ( t=3.362, 5.122, P<0.05), and recovered with overexpression of Cx43 ( t=2.674, 4.398, P<0.05). Conclusions:X-ray irradiation leads to the phosphorylation of Ser368 at Cx43, which promotes the degradation and nucleus/perinuclear translocation of Cx43 and reduces the stiffness of HUVEC. Increasing the expression level of Cx43 is helpful to the stiffness recovery of irradiated vascular endothelial cells, suggesting that Cx43 may be a potential target for regulating radiation injury of vascular endothelial cells.

OBJECTIVE: Pyroptosis is an inflammatory form of programmed cell death. This phenomenon has been recently reported to play an important role in radiation-induced normal tissue injury. Connexin43 (Cx43) is a gap junction protein that regulates cell growth and apoptosis. In this study, we investigated the effect of Cx43 on X-ray-induced pyroptosis in the human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs, Cx43 overexpression, and Cx43 knockdown strains were irradiated with 10 Gy. Proteins were detected using western blot analysis. Cell pyroptosis was evaluated using the fluorescence-labeled inhibitor of caspase assay (FLICA) and propidium iodide staining through flow cytometry and confocal microscopy. Cell morphology and cytotoxicity were detected by scanning electron microscopy and lactate dehydrogenase release assay, respectively. RESULTS: Irradiation with 10 Gy X-ray induced pyroptosis in the HUVECs and reduced Cx43 expression. The pyroptosis in the HUVECs was significantly attenuated by overexpression of Cx43 as it decreased the level of active caspase-1. However, interference of Cx43 expression with siRNA significantly promoted pyroptosis by increasing the active caspase-1 level. Pannexin1 (Panx1), a gap junction protein regulates pyroptosis, and its cleaved form is used to evaluate channel opening and active state. The level of cleaved Panx1 in the HUVECs and Cx43 knockdown strains increased in the presence of X-ray, but decreased in the Cx43 overexpression strains. Furthermore, interference of Panx1 with siRNA alleviated the upregulation of pyroptosis caused by Cx43 knockdown. CONCLUSION Results suggest that single high-dose X-ray irradiation induces pyroptosis in the HUVECs. In addition, Cx43 regulates pyroptosis directly by activating caspase-1 or indirectly by cleaving Panx1.
Caspase 1 ; genetics ; metabolism ; Connexin 43 ; genetics ; metabolism ; Connexins ; genetics ; metabolism ; Gene Expression Regulation ; radiation effects ; Human Umbilical Vein Endothelial Cells ; physiology ; radiation effects ; Humans ; Nerve Tissue Proteins ; genetics ; metabolism ; Pyroptosis ; X-Rays ; adverse effects

The present study cloned wpGuamerin gene from a non-bloodsucking leech (Whitmania pigra), and the bioinformatics analysis of the sequence was performed. Additionally, the effects of feeding duration on the expression profile of the wp Guamerin gene were explored. The results showed that its sequence consisted of 295 nucleotides encoding a peptide of 83 amino acids(Genbank: KX768545), and its relative molecular weight is 9 145.95 Da. wp Guamerin does not encode proteins with a signal peptide, belonging to the hydrophilic protein. Its secondary structure is mainly composed of α-helix, extending chain, folding and random curl. Its similarity with other blood-sucking leeches ranges from 29% to 65%. The results revealed that wpGuamerin mRNA was detected higher expression in muscle than in salivary glands of Wh. pigra, and did not expressed in ingluvies and intestine. Its expression in muscle and salivary glands showed a single peak curve after feeding and the peak was observed in the 1st and 3rd after feeding, respectively. In summary, wp Guamerin in Wh. pigra is a small molecule polypeptide protein and is different from the Guamerin in blood-sucking leeches. wpGuamerin does not express in the digestive tract of Wh. pigra, and mainly express in muscle. Feeding behavior would stimulate the expression of wpGuamerin gene in muscle and salivary glands, but not in digestive tract.

OBJECTIVETo explore the methods and results of modified one-stage revision procedure for treating proximal femoral infected nonunion after intramedullary nail fixation.

METHODSFrom June 2010 to June 2015, 10 patients of proximal femoral infected nonunion after intramedullary nail fixation were treated with modified one-stage revision procedure, including 9 males and 1 female, ranging in age from 35 to 77 years old. There were 3 cases of intertrochanteric fractures, 2 cases of intertrochanteric fractures accompanied with proximal femoral fractures and 5 cases of subtrochaneric fracures. The fractures ware fixed by LISS plate after radically debridement. The bone defects were repaired by free vascularized fibular graft and autogenous cancellous bone graft mixed artificial bone containing antibiotics. Postoperatively, ambulation without weight bearing was encouraged as early as possible.

RESULTSTen patients were followed up from 9 to 30 months and all nonunions healed smoothly without wound infection recurrence or internal fixation failure at the final follow-up. The time for full weight bearing was from 12 to 28 weeks. The hip joint function was evaluated by Sanders Traumatic Hip Rating Scale, the result was excellent in 7 cases, good in 2, and fair in 1 at the final follow-up.

CONCLUSIONSModified one-stage revision procedure is an effective treatment with a good functional result for proximal femoral infected nonunion after intramedullary nail fixation. On the basis of radical debridement, the combination of infection control and bone healing therapeutic techniques is key for success.

Objective To study the role of Cx43 in X-ray induced apoptosis of HUVEC cells and its mechanism.Methods Flow cytometry was used to detect the apoptosis of HUVEC cells at 48-96 h after 10 Gy X-ray irradiation and at 72 h after irradiation of different doses.Western blot was used to detect the protein expressions of Cx43 and cleaved caspase-3 in HUVEC cells at 72 h after 0,5,10 and 20 Gy irradiation.Small interfering RNA was transfected into HUVEC cells to silence Cx43 expression,the Cx43 bearing plasmid was transfected into cells to overexpress Cx43.The effect of Cx43 knockdown or overexpression on apoptosis induction and cleaved caspase-3 protein expression were detected by flow cytometry and Western blot,respectively.Results The apoptosis of HUVEC increased significantly from 48 h to 96 h after X-ray irradiation and in a dose-dependent manner at 72 h after irradiation.The expression of Cx43 protein was negatively correlated with the dose but the expression of cleaved caspase-3 was positively correlated with the dose in the range of 0-20 Gy.After Cx43 silencing,the proportion of early apoptosis and apoptosis combined with dead cells were significantly higher than that of the siRNA control group(t =3.674,6.375,P ＜ 0.05).After Cx43 overexpression,the proportion of early apoptosis and apoptosis combined with dead cells were significantly lower than that of vector control group(t =9.399,11.190,P ＜ 0.05).The expression of cleaved caspase-3 in the Cx43 silencing group was higher than that in the siRNA control group,but this protein in the Cx43 overexpressed group was lower than that in the vector control group.Conclusions Cx43 may protect X-ray irradiated HUVEC cells from apoptosis by down-regulating the activation of caspase-3.

OBJECTIVETo evaluate the middle-long-term clinical effects of unilateral and bilateral percutaneous kyphoplasty(PKP) for vertebral fragility fracture in the elderly.

METHODSThe clinical data in elderly patients with vertebral fragility fracture treated by unilateral and bilateral PKP between January 2008 and January 2010 was retrospective analyzed. According to a unified criteria to divided into two groups for 104 patients(44 males and 60 females), of them, 50 cases were divided in unilateral group using unilateral pedicle surgical approach for PKP and 54 cases were in bilateral group using bilateral pedicle approach. VAS score, Cobb angle, and the height of anterior and posterior vertebral body were respectively analyzed peroperatively and at 3 d, 3 months, 1 year, 3 years postoperatively and final follow-up. Clinical effects and safety were assessed in two groups.

RESULTSAll the operations were successful. Operative time and bone cement injection volume in unilateral group were less than those of bilateral group(<0.05). Postoperative VAS scores, Cobb angle, and the height of anterior and posterior vertebral body were obviously improved in two groups(<0.05), and there was no significant difference between two groups(>0.05). Bone cement leakage occurred in 12 cases(11.5%), recurrent fracture of vertebral body occurred in 5 cases(4.8%), cerebrospinal leak occurred in 2 cases(1.9%), and nerve root irritation occurred in 3 cases(2.9%). The above complications were transient and released after symptomatic treament.

CONCLUSIONSMiddle-long-term clinical effects of unilateral and bilateral percutaneous kyphoplasty for vertebral fragility fracture are safe and satisfactory, and may be extended as a minimally invasive method. Unilateral approach for PKP has advantages of short operation time, relative less trauma, thus is a more ideal method.