Objective:To evaluate the effects of intrathecal infusion chemotherapy on intracranial pressure (ICP) in non-small cell lung cancer (NSCLC) patients with leptomeningeal metastases (LM) by ultrasound measurement of the optic nerve beside the bed of optic nerve sheath diameter (ONSD) .Methods:A total of 31 NSCLC-LM patients who underwent intrathecal infusion chemotherapy at Nanjing Drum Tower Hospital, Affiliated Hospital of Nanjing University Medical School from June 10, 2021 to December 25, 2022 were collected. The ONSD values were measured before and after the first lumbar puncture by bedside optic nerve ultrasound, and measured dynamically 30 min before intrathecal infusion chemotherapy (T0) , 30 min (T1) , 1 h (T2) , 2 h (T3) , 4 h (T4) , 6 h (T5) , and 24 h (T6) after intrathecal infusion chemotherapy. ICP ONSD was calculated, with differences between ICP LP and ICP ONSD, and differences between ONSD and ICP ONSD series at different time being compared separately. Mean arterial pressure (MAP) , heart rate, and headache score were assessed and compared respectively at T0, T1, T2, T3, T4, T5 and T6. Spearman analysis was used to evaluate the correlation between the response assessment in neuro-oncology (RANO) score and ICP. Results:Before the first lumbar puncture for cerebrospinal fluid drainage, ICP LP was (218.55±63.83) mmH 2O, left eye, right eye, and binocular eyes ICP ONSD were (217.28±57.17) mmH 2O, (223.64±51.13) mmH 2O, and (220.46±52.50) mmH 2O respectively, in NSCLC-LM patients, with no statistically significant difference ( F=0.77, P=0.463) . After first lumbar puncture for cerebrospinal fluid drainage, ICP LP was (214.68±58.01) mmH 2O, left eye, right eye, and binocular eyes ICP ONSD were (216.71±48.96) mmH 2O, (216.62±47.18) mmH 2O, and (216.67±47.86) mmH 2O respectively, with no statistically significant difference ( F=0.12, P=0.757) . At T0, T1, T2, T3, T4, T5, and T6, the MAP during intrathecal infusion chemotherapy was 89.80 (83.40, 93.67) mmHg, 95.00 (80.83, 99.37) mmHg, 91.86 (79.88, 100.14) mmHg, 90.15 (79.04, 100.55) mmHg, 105.14 (88.55, 114.74) mmHg, 98.96 (81.72, 111.81) mmHg, and 89.29 (85.45, 100.38) mmHg, with a statistically significant difference ( χ2=16.11, P=0.013) ; heart rates were 80.00 (75.00, 84.50) times/min, 80.00 (72.50, 87.50) times/min, 74.00 (66.00, 87.50) times/min, 82.00 (72.00, 90.00) times/min, 80.00 (70.50, 90.00) times/min, 77.00 (68.00, 91.00) times/min, 77.00 (71.50, 88.50) times/min, with no statistically significant difference ( χ2=2.18, P=0.902) ; headache scores were 2.00 (0.50, 3.00) score, 2.00 (1.00, 3.00) score, 2.00 (2.00, 3.00) score, 2.00 (1.00, 3.00) score, 2.00 (1.00, 2.00) score, 2.00 (1.00, 2.00) score, and 2.00 (0.00, 2.00) score, with no statistically significant difference ( χ2=11.64, P=0.071) . At T0, T1, T2, T3, T4, T5, and T6, left eye, right eye, and binocular ONSD were (5.85±0.64) mm, (5.72±0.68) mm, (7.11±1.11) mm, (6.42±0.78) mm, (5.69±0.63) mm, (5.61±0.64) mm, (5.65±0.88) mm, (5.85±0.12) mm, (5.89±0.12) mm, (6.93±0.20) mm, (6.40±0.14) mm, (5.71±0.12) mm, (5.66±0.12) mm, (5.33±0.14) mm, (5.85±0.64) mm, (5.81±0.64) mm, (7.02±1.03) mm, (6.41±0.75) mm, (5.70±0.63) mm, (5.64±0.63) mm, (5.49±0.76) mm, with statistically significant differences ( F=58.48, P<0.001; F=49.34, P<0.001; F=78.05, P<0.001) ; ICP ONSD were (222.81±56.81) mmH 2O, (211.89±60.29) mmH 2O, (335.12±98.32) mmH 2O, (274.17±68.87) mmH 2O, (208.77±56.12) mmH 2O, (201.75±56.79) mmH 2O, (205.59±78.36) mmH 2O, (223.26±58.33) mmH 2O, (227.08±61.68) mmH 2O, (319.36±101.10) mmH 2O, (272.33±69.61) mmH 2O, (211.21±57.73) mmH 2O, (206.51±57.22) mmH 2O, (177.22±68.98) mmH 2O, (223.03±57.24) mmH 2O, (219.49±57.24) mmH 2O, (327.24±91.56) mmH 2O, (273.25±67.04) mmH 2O, (209.99±56.26) mmH 2O, (204.13±56.29) mmH 2O, (191.40±67.95) mmH 2O, with statistically significant differences ( F=58.48, P<0.001; F=49.34, P<0.001; F=78.13, P<0.001) . The ONSD of the left eye, right eye, and binocular eyes and the corresponding ICP ONSD increased significantly at T2 compared with T0, T1, T3, T4, T5, and T6, with statistically significant differences (all P<0.05) . Pre- and post-treatment RANO scores were 4.00 (3.00, 7.00) score and 3.00 (2.00, 6.00) score respectively. Pre- and post-treatment RANO scores were positively correlated with ICP ONSD in the left eye ( r=0.55, P=0.001; r=0.60, P<0.001) , right eye ( r=0.54, P=0.001; r=0.46, P=0.009) and binocular eyes ICP ONSD ( r=0.45, P=0.010; r=0.37, P=0.043) . Conclusion:Intrathecal infusion chemotherapy for NSCLC-LM patients can cause a transient increase in ONSD and ICP, with the greatest effect at 1 hour after intrathecal infusion chemotherapy. RANO score is positively correlated with ICP ONSD before and after treatment, which can provide an important reference for evaluating the efficacy of intrathecal infusion chemotherapy.

Objective:To investigate the pharmacokinetics of cerebrospinal fluid pemetrexed following intrathecal injection chemotherapy in patients with leptomeningeal metastasis (LM) from lung adenocarcinoma and provide a basis for clinical intrathecal injection chemotherapy.Methods:A total of 21 patients with lung adenocarcinoma LM who underwent pemetrexed intrathecal injection chemotherapy via Ommaya capsule at Nanjing Drum Tower Hospital, Aiffilitated Hospital of Nanjing University Medical School from November 2019 to November 2022 were collected, and divided into 30, 40 and 50 mg groups ( n=10, n=4, n=7) according to pemetrexed dose. Cerebrospinal fluid was collected at 0, 0.5, 1, 2, 4, 6, 12, 24 and 48 h after the first intrathecal injection chemotherapy, and day 8 of each cycle for three groups. Reversed phase high performance liquid chromatography was used to determine the drug concentration in cerebrospinal fluid, to clarify the drug-related pharmacokinetic parameters, and to compare the differences in pemetrexed concentration among groups. Finally, cerebrospinal fluid pemetrexed concentration changes were observed and compared after different intrathecal injection chemotherapy cycles. Results:There were statistically significant differences in cerebrospinal fluid drug concentrations of patients in three groups at 0, 0.5, 1, 2, 4, 6, 12, 24 and 48 h after the first intrathecal injection chemotherapy (30 mg group: F=20.56, P<0.001; 40 mg group: F=27.06, P<0.001; 50 mg group: F=28.63, P<0.001), and there were statistically significant differences in the concentration of cerebrospinal fluid drugs in each dose group at 0.5, 1, 2, 4, 6 and 12 h compared to 0 h after intrathecal injection chemotherapy (all P<0.05). Compared to the 30 mg group, cerebrospinal fluid drug concentrations in the 50 mg group increased at 1, 2, 4, 6, 12 and 24 h after intrathecal injection chemotherapy, with statistically significant differences (all P<0.05). Pharmacokinetic analysis of cerebrospinal fluid pemetrexed showed that area under the concentration-time curve (AUC) 0-∞ of the 30, 40 and 50 mg groups were (5 696.12±283.32), (7 886.29±396.57), and (14 202.70±440.19) h·mg/L, respectively, with a statistically significant difference ( F=1 159.00, P<0.001) ; AUC 0-∞ increased in the 50 mg group compared to the 30 and 40 mg groups (both P<0.05) ; AUC 0-∞ increased in the 40 mg group compared to the 30 mg group ( P<0.05). The half-lives of three groups were (8.75±0.23), (11.29±0.59) and (16.42±1.23) h, respectively, with a statistically significant difference ( F=206.80, P<0.001) ; half-life was longer in the 50 mg group compared to the 30 and 40 mg groups (both P<0.05) ; half-life was longer in the 40 mg group compared to the 30 mg group ( P<0.05). The peak time of three groups were (1.55±0.10), (1.00±0.01), (1.43±0.11) h, respectively, with a statistically significant difference ( F=48.11, P<0.001) ; the peak time was shorter in the 40 and 50 mg groups compared to the 30 mg group (both P<0.05). Clearance of three groups were (7.02±2.46), (5.80±1.25) and (3.66±1.32) L/h, respectively, with a statistically significant difference ( F=6.02, P=0.009) ; clearance was decreased in the 50 mg group compared to the 30 mg group ( P<0.05). The peak concentration of three groups were (540.45±32.25), (820.75±46.47) and (1 014.78±64.96) mg/L, respectively, with a statistically significant difference ( F=207.70, P<0.001) ; peak concentration increased in the 50 mg group compared to the 30 and 40 mg groups (both P<0.05) ; peak concentration increased in the 40 mg group compared to the 30 mg group ( P<0.05). Cerebrospinal fluid drug concentrations were dynamically monitored after 4 cycles of intrathecal injection chemotherapy, in which cerebrospinal fluid pemetrexed concentrations in 30 mg group were (13.76±4.79), (11.41±7.08), (9.41±2.59) and (7.86±4.02) mg/L, respectively; 40 mg group were (14.45±6.59), (12.87±15.73), (11.24±2.48) and (9.09±3.38) mg/L, respectively; 50 mg group were (12.94±10.34), (9.72±7.62), (8.15±8.17) and (4.34±4.21) mg/L, respectively. There was a statistically significant difference in cerebrospinal fluid drug concentrations among different intrathecal injection chemotherapy cycles in 30 mg group ( F=4.04, P=0.016), and the cerebrospinal fluid drug concentration decreased in cycles 3 and 4 compared to cycle 1 (both P<0.05). There were no statistically significant differences in cerebrospinal fluid drug concentrations among different treatment cycles in 40 and 50 mg groups ( F=0.28, P=0.837; F=3.57, P=0.066) . Conclusion:Reversed phase high performance liquid chromatography method can effectively detect the pemetrexed concentration in cerebrospinal fluid; dynamic monitoring of cerebrospinal fluid pemetrexed concentration can provide a basis for the dosage and the treatment cycle of intrathecal injection chemotherapy in LM patients with lung adenocarcinoma.

Objective To investigate the effect of transcription factor specific protein5(Sp5)silencing on Wnt signaling pathway correlated factors and cell proliferation ability in mouse embryo palatal mesenchymal cells(mEPMCs).Methods mEPMCs of 14.5 d pregnant C57BL/6J mice were isolated and cultured in vitro.Cell source was identified by immunofluorescence staining.Lentivirus transfection technique was used to silence the expression of Sp5 gene in mEPMCs,and the transfection efficiency was verified by Western blot assay.Follow-up experiments were set up with the blank control group,the no-load virus group and the slience group(the Sp5-shRNA group).The protein and mRNA expression levels of β-catenin,GSK-3β,Wnt3a and CyclinD1 were detected by Western blot assay and RT-qPCR after transfection for 72 h in each group.Cell proliferation capacity was detected by CCK-8.The proliferation rate of 5-Ethynyl-2'-deoxyuridine(EdU)positive cells was detected by immunofluorescence assay.Cell cycle was detected by flow cytometry.Results mEPMCs were successfully isolated,and Sp5 expression was silenced.Western blot and RT-qPCR results showed that the protein and mRNA expressions of β-catenin,GSK-3β,Wnt3a and CyclinD1 were significantly higher in the Sp5-shRNA group than those in the blank control group and the no-loaded virus group(P<0.05).The proliferative ability and the proliferative rate of EdU positive cells were higher in the Sp5-shRNA group than those in the blank control group and the no-loaded virus group(P<0.05).The proportion of mEPMCs in S phase was higher in the Sp5-shRNA group than that in the blank control group and the no-loaded virus group(P<0.05).Conclusion Sp5 in silenced mEPMCs can participate in palate development and promote the proliferation of mEPMCs by regulating Wnt signaling pathway.

Objective　To investigate the effect of Salvianolic acid on the expression of glial fibrillary acidic protein (GFAP),protein gene product (PGP9.5),IL-5,IL-6,IL-8,IL-10 and prognosis in patients with acute ischemic stroke.Methods　84 patients with acute ischemic stroke were divided into salvia miltiorrhiza group (n=42) and control group (n=42).Barthel index and NIHSS score were performed on the day of admission and 14 days.The post-infarction volume was calculated and compared after head MRI.The expression levels of serum GFAP,PGP9.5,IL-5,IL-6,IL-8 and IL-10 were detected and compared by magnetic particle chemiluminescence method.The prognosis was determined by modified Rankin score (mRS) 90 days after infarction.Results　After treatment,NIHSS score,Barthel index and infarct volume in salvia miltiorrhiza group were significantly decreased (P<0.05),significantly increased (P<0.05) and significantly decreased (P<0.05).GFAP,IL-6 and IL-8 decreased significantly in salvia miltiorrhiza group after treatment compared with the control group (all P<0.05).After treatment,IL-10 expression level in salvia miltiorrhiza group increased more significantly than that in control group (P<0.05).The prognosis of Salvia miltiorrhiza group was better than that of control group (P<0.05).Conclusion　Salvianolic acid in the treatment of acute ischemic stroke can effectively inhibit inflammatory reaction,reduce brain injury,improve neurological impairment,improve daily behavior and improve prognosis.

Objective To evaluate the feasibility and clinical efficacy of our self-designed simple skin stretching device combined with collagen sponge for management of severe soft tissue wounds.Methods From September 2015 to October 2017,a consecutive series of 43 patients whose soft tissue wounds could not be closed primarily were enrolled for a therapy using a simple skin stretching device made of round osseous pins and wire combined with collagen sponge.They were 27 males and 16 females,with a mean age of 31.5 years (from 5 to 56 years).There were 18 fresh wounds and 25 old ones.Their skin defects ranged from 5.5 cm × 3.0 cm to 18.0 cm × 7.5 cm.After debridement and vacuum sealing drainage,2 round osseous pins with a diameter of 2.0 mm or 2.5 mm were driven through the dermis about 1 to 2 cm from both edges of the wound,in parallel with the longitudinal axis of the wound.After the parts of 2 pins exposed outside the skin were bent,they were fixed respectively with a fine wire with 2 twisted strands.The wounds were continuously stitched with eversion suture.The wires and sutures were gradually tightened to contract the wounds until the skin color changed and capillary filling reaction started.Then medical collagen sponge was used to cover the wounds.Next,the wires and sutures were tightened continuously until the wound edges were pulled together.Details of this therapy and its complications were recorded.Follow-up visits were paid until wound healing.Results Of the 43 cases,the wounds were directly closed immediately after primary stretching procedure in 8,closed after skin stretching for 4 to 12 days (average,7.5 days) in 30,and significantly reduced in 5 which were cured following skin graft.Eventually,40 cases were followed up for an average of 6.8 months (from 3 to 18 months) and 3 were lost.Aesthetic reoperation was performed in 3 patients who were inflicted with postoperative scar formation after skin graft.Linear healing of the wound edges was achieved in 37 patients without complications like skin necrosis,pathological hyperplasia scar,skin sensation deletion or wound infection,leading to fine appearance and functional recovery.Conclusion Our self-designed simple skin stretching device combined with collagen sponge provides a cost-effective and practical technique for clinical treatment of soft tissue defects,with an advantage of reducing or even avoiding secondary repair with skin graft or skin flap.

OBJECTIVETo identify potential mutation in a Chinese family affected with Charcot-Marie-Tooth disease(CMT).

METHODSClinical data of the family was collected, and genomic DNA was extracted from peripheral blood samples of the family members. Seventy-two candidate genes of the proband were captured and sequenced by targeted next-generation sequencing, and the results were confirmed by Sanger sequencing. The protein structure was predicted with PyMOL-1 software.

RESULTSA homozygous missense mutation c.1894G>A(p.E632K) was identified in the exon 11 of the SH3TC2 gene in the proband. Heterozygous c.1894G>A mutation was also detected in the proband's father, mother and daughter, but not in the healthy family members and 300 normal controls. Retrieval of the NCBI, HGMD and 1000 genome databases has verified the c.1894G>A to be as a novel mutation. Computer simulation has suggested that the mutation has altered the 3D structure of the SH3TC2 protein.

CONCLUSIONThe proband was diagnosed as CMT4C, for which the underlying gene was SH3TC2. This finding has expanded the spectrum of SH3TC2 mutation in association with CMT4C.

Adult ; Aged ; Base Sequence ; Charcot-Marie-Tooth Disease ; genetics ; DNA Mutational Analysis ; Exons ; Female ; Heterozygote ; Humans ; Male ; Middle Aged ; Models, Molecular ; Molecular Sequence Data ; Mutation, Missense ; Pedigree ; Proteins ; genetics ; Young Adult

Druggability is crucial in pharmaceutical drug development as the source of drug research. Druggability research will face greater challenges because Chinese materia medica (CMM) is the multicomponent drug. In this paper, ideas and methods of study on CMM druggability were mainly proposed in combination with the chemical material basis of muticomponents of CMM.

Objective To explore the variation trends of interleukin-1β( IL-1β) and intercellular adhesion molecule-1 (ICAM-1) in both normal and affected sides of brain tissues in rats with ischemia-reperfusion injury and the therapeutic action of electroacupuncture.Methods The cerebral ischemia-reperfusion model was established with suture embolization in the right middle cerebral artery.The rats were randomly divided into control group, model group and electroacupunture group.Each group was then divided into six subgroups by the time after operation (12 h,24 h,48 h,72 h,96 h,144 h), ten rats in each subgroup. Frozen sections of brain tissues were prepared and the expression of IL-1βand ICAM-1 in brain tissues of both sides were detec-ted by immunohistochemistry.Results The expressions of IL-1βand ICAM-1 showed typical bimodal pattern in both affected is-chemic region and contralateral normal region.In the model group, the peaks of IL-1βin the cerebral ischemic region were at 12 h and 48 h, while in the contralateral normal region the peaks were at 12 h and 144 h, the expression of IL-1βin the ischemic region was significantly higher than that in the contralateral normal region at 48 h (P<0.05), and lower at 96 h and 144 h (P <0.05).In the electroacupuncture group, the expressions of IL-1βin the ipsilateral region were significantly lower than that in the contralateral region at 24 h, 48 h and 144 h (P<0.05).In the model group, the peaks of ICAM-1 in the cerebral ischemic regions were at 24 h and 72 h, while in the contralateral normal regions the peaks were at 24 h and 144 h.In the electroacupunc-ture group, the expressions of ICAM-1 in the ischemic regions were significantly lower than that in the contralateral normal re-gions at all the 12 h, 24 h, 48 h, 72 h and 144 h (P<0.05).Conclusions Our findings suggest that electroacupuncture may inhibit the inflammation of ischemia/reperfusion brain tissue through reducing the expression of IL-1βand ICAM-1 to relieve the cerebral ischemia-reperfusion injury.

Objective To investigate the effects of Rhizoma Alismatis extracts on oxidative stress induced by cerebral ischemia-reperfusion injury in rats,and to explore its protective mechanism in cerebral ischemia-reperfusion injury.Methods A total of 60 male SD rats were randomly divided into sham operation group,model group,Alisma orientalis group and Nimodipine positive control group (n=15,each).Cerebral ischemia-reperfusion injury model was prepared by suture method after 14 days of intragastric administration.After 24 hours,scores of neurological dysfunction,the infarct size,the water content of the brain,the malondialdehyde (MDA),superoxide dismutase (SOD),nitric oxide (NO) levels in serum and brain tissues,and the activity of inducible NO synthase (iNOS)were detected.Results As compared with the model group,Alisma orientalis group showed that the scores of neurological dysfunction,cerebral water content,cerebral infarction size,contents of MDA and NO,and the activity of iNOS were significantly reduced,and the activity of SOD was significantly increased in respectively [(2.21 ± 0.38) vs.(2.78 ± 0.43),(81.18 ± 2.09)％ vs.(88.33±4.15)％,(0.26±0.07) ％ vs.(0.35±0.04)％,(5.92±1.64) μmol/L vs.(8.21±1.47)μmol/L,(115.48±18.65) mU/L vs.(75.52±20.78) mU/L,(28.23±4.32) μmol/L vs.(41.73±3.85) μmol/L,(15.31±1.68) mU/L vs.(23.49±3.53) mU/L,(5.41±0.68) μmol/L vs.(7.58±1.49) μmol/L,(168.57±10.65) mU/L vs.(150.11±13.62) mU/L,(14.37±0.77) μmol/L vs.(22.08±1.57) μmol/L,(9.83±0.75) mU/L vs.(13.28±1.84) mU/L,respectively,all P＜0.05]Conclusions Alisma orientalis extract has the protective effect on focal cerebral ischemia reperfusion injury,and the mechanism may be related to antioxidant and scavenging free radicals.

Objective To investigate and analysis the situation of angiotensin-converting enzyme (ACE) gene polymorphism in military service officers of Jinan theater .Methods Roche Cycler480II fluorescence quantitative PCR analyzer was used to detected ACE genotype .Meanwhile ,some samples were randomly analyzed by agarose gel electrophoresis .Results ACE DD genotype ac-counted for 23 .3% in the military service officers of Jinan theater cadres ,ID type accounted for 43 .2% ,II type accounted for 33 .5% ,D and I allele frequencies were 0 .44 and 0 .56 ,respectively ;type II ACE gene frequency was highest in female cadres ,while frequency of ID type was highest in male cadres ,the difference was statistically significant between two groups (P< 0 .05);the difference of allele frequency between men and women was not statistically significant (P>0 .05) .Conclusion ACE gene polymor-phism with type II and Iallele were dominant in the military service officers of Jinan theater cadres ;ACE gene polymorphism survey is important for early detection and timely prevention of certain related diseases .