BackgroundDry eye is a multi-factorial-induced tear film and ocular surface disorder.Immunoinflammation plays a key role in the pathogenesis of dry eye.As inhibitor of the cyclo-oxygenase pathway,nonsteroidal anti-inflammatory drugs play an anti-inflammatory and anti-hypersensitivity role,and it can be a potential treatment for dry eyes.ObjectiveThis study was to investigate the effectiveness of nonsteroidal anti-inflammatory drugs (0.1％topical pranoprofen) on moderate to severe dry eyes and its mechanism.MethodsThis was a small sample of randomized controlled clinical trial.Thirty right eyes of 30 patients with moderate to severe dry eyes were included in the study according to the diagnosis criteria and randomized into two groups.The patients of the trial group received topical administration of 0.1％ pranoprofen plus 0.1％ sodium hyaluronate,and those of the control group received the topical 0.1％ sodium hyaluronate only.Ocular surface inflammation index scores (OSDI) and ocular surface fluorescine staining (OSS) scores were measured under the slit lamp,and tear film break-up time (BUT),Schirmer Ⅰ test values were evaluated.The expression of human leucocyte antigen-DR (HLA-DR) and CD11b in conjunctiva epithelial cells were detected by impression cytology and flow cytometry (FCM).All the indexes were compared between the two groups before and after treatment.Informed consent was obtained from all patients.ResultsThere were no significant differences in terms of age and gender and their baseline values between the trial group and control group (t=0.412,P=0.684;x2=0.240,P=0.624),and so were all the indexes (P＞0.05).Compared with the control group,the OSDI,OSS scores and cells positive for HLA-DR were lowered but the BUT was delayed in the trial group on day 15 ( t=2.43,P=0.03;t=2.83,P=0.01;t=3.29,P=0.00;t=3.23,P=0.00 ).No significant differences were found in the Schirmer Ⅰ test value and CD11b expression between these two groups (t=0.17,P=0.87;t=0.28,P=0.79).The OSDI,OSS scores and BUT were significantly improved,and the number of cells positive for HLA-DR were reduced 15 days after administration of drugs in comparison with before treatment in the trial group ( t =12.30,10.70,6.10,7.92,P =0.00 ).However,there were no comparable alteration seen in these indexes before and after the usage of drugs in the control group ( P＞0.05).Positive correlations were found in HLADR expression with OSDI and OSS ( r =0.601,P =0.018 ; r =0.586,P =0.022 ) and a negative correlation in HLADR expression with BUT (r=-0.697,P=0.004) on day 15 in the trial group.ConclusionsTopical usage of 0.1％ pranoprofen is beneficial for remitting the ocular signs and symptoms in moderate to severe dry eyes.This study illustrates that topical usage of 0.1％ pranoprofen can down-regulate the expression of inflammatory markers in conjunctival epithelial cells.

OBJECTIVETo explore the mechanism of electroacupuncture scalp point penetration therapy in treatment of the Parkinson's disease (PD).

METHODSForty Wistar rats were randomly divided into a normal group, a sham-operation group, a model group and an electroacupuncture (EA) group. 6-OHDA was injected into the left striatum to make lateralization PD rat model. Acupuncture at "Baihui" (GV 20)-through-"Taiyang" (EX-HN 5), once each day, 6 days constituting one course. Immunohistochemical method was used to observe the facio-density and the integral optical density of brain-derived neurotrophic factor (BDNF) in the left substantia nigra, and TUNEL method was used to observe the apoptotic amount, and high performance liquid chromatography was used to observe DA contents of the left striatum in each group.

RESULTSAs compared with the model group, in the acupuncture group, the facio-density and the integral optical density in the left substantia nigra increased significantly (P < 0.05), the amount of apoptosis decreased significantly (P < 0.05), and the content of striatum DA increased significantly (P < 0.05).

CONCLUSIONEA scalp point-through-point therapy may enhance BDNF protein expression level in the substantia nigra to decrease the amount of apoptosis in the PD model rat.

Acupuncture Points ; Animals ; Apoptosis ; Brain-Derived Neurotrophic Factor ; metabolism ; Electroacupuncture ; methods ; Female ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Oxidopamine ; administration & dosage ; toxicity ; Parkinson Disease, Secondary ; chemically induced ; therapy ; Random Allocation ; Rats ; Rats, Wistar ; Scalp ; pathology ; Substantia Nigra ; metabolism ; pathology ; Treatment Outcome

BACKGROUNDIsoflurane, a commonly used inhaled anesthetic, induces apoptosis in primary rat cortical neurons of rat in a concentration- and time-dependent manner by an unknown mechanism. We hypothesized that isoflurane induced apoptosis by causing abnormal calcium release from the endoplasmic reticulum (ER) via activation of inositol 1, 4, 5-trisphosphate (IP(3)) receptors. Sevoflurane has a reduced ability to disrupt intracellular calcium homeostasis and is a less potent cytotoxic agent. This study examined and compared the cytotoxic effects of isoflurane and sevoflurane on rat primary cortical neurons and their relationship with disruption of intracellular calcium homeostasis and production of reactive oxygen species (ROS).

METHODSPrimary rat cortical neurons were treated with the equivalent of 1 minimal alveolar concentration (MAC) of isoflurane and sevoflurane for 12 hours. MTT reduction and LDH release assays were performed to evaluate cell viability. Changes of calcium concentration in the cytosolic space, [Ca(2+)](c), and production of ROS were determined after exposing primary rat cortical neurons to isoflurane and sevoflurane. We also determined the effects of IP(3) receptor antagonist xestospongin C on isoflurane-induced cytotoxicity and calcium release from the ER in primary rat cortical neurons.

RESULTSIsoflurane at 1 MAC for 12 hours induced cytotoxicity in primary rat cortical neurons, which was also associated with a high and fast elevation of peak [Ca(2+)](c). Xestospongin C significantly ameliorated isoflurane cytotoxicity in primary cortical neurons, as well as inhibited the calcium release from the ER in primary cortical neurons. Isoflurane did not induce significant changes of ROS production in primary rat cortical neurons. Sevoflurane, at equivalent exposure to isoflurane, did not induce similar cytotoxicity or elevation of peak [Ca(2+)](c) in primary rat cortical neurons.

CONCLUSIONThese results suggested that isoflurane induced elevation in [Ca(2+)](c), partially via elevated activity of IP(3) receptors, which rendered cells vulnerable to isoflurane neurotoxicity. ROS production was not involved in isoflurane-induced neurotoxicity. Sevoflurane, at an equivalent exposure to isoflurane, did not induce similar elevations of [Ca(2+)](c) or neurotoxicity in primary cortical neurons of rat.

Anesthetics, Inhalation ; toxicity ; Animals ; Calcium ; metabolism ; Cell Survival ; drug effects ; Cells, Cultured ; Inositol 1,4,5-Trisphosphate Receptors ; drug effects ; physiology ; Isoflurane ; toxicity ; Methyl Ethers ; toxicity ; Rats ; Reactive Oxygen Species ; metabolism

OBJECTIVESTo identify and clone human genes transactivated by HCV F protein by constructing a cDNA subtractive library using the suppression subtractive hybridization technique.

METHODSSuppression subtractive hybridization (SSH) and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV F protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1 (-)-F or with pcDNA3.1(-) empty vector as a control, and SSH method was employed to analyze the differentially expressed DNA sequence between the two groups. After restriction enzyme Rsa I digestion, small sized cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 or adaptor 2. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, it was then subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5 alpha. The cDNA was sequenced and analyzed in GenBank with blast search after PCR.

RESULTSThe subtractive library of genes transactivated by HCV F protein was constructed successfully. The amplified library contains 71 positive clones. Colony PCR shows that 56 clones contain 200-1000 bp inserts. Sequence analysis was performed on 28 clones randomly, and the full length sequences were obtained with using the bioinformatics method. Altogether 19 coding sequences were obtained, consisting of 17 known and 2 unknown.

CONCLUSIONSThe obtained sequences may be target genes transactivated by HCV F protein, and some gene coding proteins are those involved in cell cycle regulation, metabolism, and cell apoptosis.

Cloning, Molecular ; Hepacivirus ; genetics ; Humans ; Nucleic Acid Hybridization ; methods ; RNA, Messenger ; biosynthesis ; genetics ; Transcriptional Activation ; Viral Core Proteins ; biosynthesis ; genetics

BACKGROUND:Autologous platelet rich plasma and bone marrow mesenchymal stem cells have certain effects on bone repair,but there are rare reports on the clinical treatment of long shaft fracture bone nonunion using autologous platelet rich plasma combined with bone marrow mesenchymal stem cell transplantation.OBJECTIVE:To investigate the therapeutic effect of autologous platelet rich plasma combined with bone marrow mesenchymal stem cell transplantation on long shaft fracture bone nonunion.METHODS:Forty-seven patients with long shaft fracture bone nonunion were randomly divided into two groups:monotherapy group (n=22) and combination group (n=25).In the monotherapy group,autologous bone marrow mesenchymal stem cell transplantation was performed in the bone nonunion site.In the combination group,autologous platelet rich plasma combined with bone marrow mesenchymal stem cell transplantation was implemented in the bone nonunion site.Callus score,clinical healing time,local complications and limb function grade were recorded and compared between the two groups.RESULTS AND CONCLUSION:The healing properties and limb function in the combination group were significantly superior to those in the monotherapy group [healing time:(4.2±1.5) vs.(5.6±1.1) months,P ＜ 0.05;healing rate:92％ vs.86％,P ＜ 0.05;callus score:2.74±0.36 vs.2.32±0.53,P ＜ 0.05;limb function recovery rate:77％ vs.84％,P ＜ 0.05].Complications like local skin redness or infection were not found in the two groups.In conclusion,both of the two methods can promote bone healing,but autologous platelet rich plasma combined with bone marrow mesenchymal stem cell transplantation has a better clinical effect on bone healing.

Objective To investigate the clinical outcomes of patients undergoing microsurgery for cerebellar medulloblastoma. Methods This retrospective analysis of the clinical and follow-up data involves 57 patients who received microsurgery for pathologically confirmed cerebellar medulloblastorna, and the microsurgical techniques for medulloblastoma were discussed. Results Among the 57 patients, 42 had total tumor resection, 13 had subtotal and 2 had partial resection of the tumors. Patency of the midbrain aqueduct was achieved in all the cases after the surgery. Hydrocephalus was found in 43 patients before the operation and only in 16 patients after the operation. Tumor relapse occurred in 19 patients 2 years after the operation, including 8 with implantation metastasis compromising the central nervous system and 1 patient with frontal lobe metastasis who received reoperations. The earliest tumor relapse occurred 20 days after the surgery. The 2- and 5-year postoperative survival rates in these patients were 68.4% and 49.1%, respectively. Conclusion Good therapeutic effects can be achieved with total resection of the tumors and postoperative whole brain and spinal cord radiotherapy in patients with medulloblastomas.

Objective To investigate the clinical data, risk factors for bleeding following endoscopic treatment of colorectal polyps (diameter ≥ 1.0 cm) and feature analysis for carcinogenesis of adenomatous polyps. Method It was analyzed retrospectively that the clinical data, endoscopic characteristics, risk factors for bleeding and features for carcinogenesis of adenomatous polyps in 741 patients with a total of 884 colorectal polyps (diameter ≥ 1.0 cm) of our hospital from January 1, 2014 to January 1, 2016, which were resected under endoscope. Result Univariate analysis of colorectal polyps resected under endoscope on intraoperative and delayed bleeding, we found that gender (P = 0.017), location (P = 0.011), size (P = 0.004), lobulated or not (P = 0.010), resection methods under endoscope (P = 0.029) were statistically significant deviation between the two groups; taking the polyp as observation unit,multivariate Logistic regression analysis of their clinical data and endoscopic characteristics, we found gender (P = 0.012, OR^ = 2.671, 95% CI = 1.246 ~ 5.728) was an independent risk factor, men are more prone to bleeding than women; it was a protective factor for the location of polyps is sigmoid colon compared to rectum (P = 0.011, OR^ = 0.348, 95% CI = 0.154 ~ 0.786), and it was a risk factor for the polyps whose diameters ≥ 3.0 cm compared to among 1.0 ~ 1.9 cm. Univariate analysis of the features for carcinogenesis of colorectal adenomatous polyps, we found that surface lobulated or not (P = 0.001), surface smooth or not (P = 0.017), Yamada classification (P = 0.008) were statistically significant deviation, and the logistic regression analysis identified surface lobulation (P = 0.001, OR^ = 6.556, 95% CI = 2.326 ~ 18.475) was an independent risk factor for carcinogenesis of adenomatous polyps. Conclusion Resection of colorectal polyps under endoscope is a safe treatment method. Taking the polyp as observation unit, gender is an independent risk factor of colorectal polyps resected under endoscope on intraoperative and delayed bleeding, men are more prone to bleeding than women; the larger the diameters of polyps are, the higher the bleeding rates could be. Surface lobulation of colorectal adenomatous polyps are more likely lead to carcinogenesis.

OBJECTIVETo explore the secure resection margin (RM) of hepatectomy for primary liver cancer (PLC) with the coexistence of cirrhosis or hepatitis by studying the correlations of the resected liver parenchyma volume with postoperative liver function, complication and RM clinically.

METHODSThe volume of tumor and the surrounding liver in resected liver specimen was measured and calculated in continuous 76 PLC patients prospectively, and the total liver parenchyma volume was measured and calculated using computed tomography (CT) images in former 40 patients. Under ideal circumstances, the surrounding liver volume, which would be resected theoretically, was calculated according to various sizes of tumors and RMs. The correlations of the resected liver volume or hepatic parenchyma-resected rate (HPRR) with postoperative liver function, complication and RM were analysed.

RESULTSThe RM was (5 +/- 7) mm in 76 patients. The volume of the tumors and the surrounding liver in the specimens were (107 +/- 203) cm(3) and (153 +/- 120) cm(3), respectively. In 40 patients, the total nontumorous liver volume using CT images was (1079 +/- 179) cm(3), and HPRR was (14 +/- 9)%. There were statistically significant differences in HPRR (P < 0.05) between three groups with complication score 0, 1-2 and 3-6 points, the value of the first group were lower than that of the third group at the level P < 0.05. The significant factors affecting liver function and complication are HPRR, the size of operation, the time of hepatic portal occlusion and the resected liver volume (P < 0.05) apart from preoperative liver function.

CONCLUSIONSWhen hepatectomy was performed in PLC patients with preoperative liver function of Child A grade and the coexistence of cirrhosis or hepatitis, 30% HPRR was a lower limit for greatly increasing the chance of developing serious postoperative complications, while 20% HPRR was a safe upper limit for achieves quick postoperative recovery or developing only a few mild complications. When PLC patients without macroscopic tumor thrombi or macrosatellites undergo hepatectomy, 10 mm RM is enough to ensure sufficient liver function residue and achieve complete micrometastasis clearance in liver parenchyma surrounding the lesion if the diameter of a tumor is less than 10 cm and 6 mm RM is enough to ensure sufficient liver function residue and obtain 99% micrometastasis clearance if the diameter of a tumor is greater than 10 cm, while with macroscopic tumor thrombi or macrosatellites, 20 mm RM is enough to ensure sufficient liver function residue and achieve 99% micrometastasis clearance if the diameter of a tumor is less than 6 cm.

Adult ; Aged ; Female ; Hepatectomy ; methods ; Hepatitis ; complications ; Humans ; Liver ; pathology ; Liver Cirrhosis ; complications ; Liver Neoplasms ; complications ; diagnostic imaging ; pathology ; surgery ; Male ; Middle Aged ; Prospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

Lignans are important defensive compounds in plants and have good biological activities protecting human health. In order to study the medicinal secondary metabolism of Fagopyrum cymosum (Trev.) Meisn, a traditional Chinese medicine with anti-tumor effect, a novel isoflavone reductase-like gene, FcIRL, was cloned using RACE strategy from a cDNA library of high flavonoids-producing callus. The full-length cDNA of the FcIRL was 1 217 bp (accession no. EU116032), which contained a 942 bp open reading frame (ORF) encoding a 313 amino acid protein. Two stop codons (TAG) and a putative polyadenylation signal ATAAA at 24 bp upstream from the polyadenylation site was found in 5' and 3' UTR, separately. And no intron was found in the genomic sequence yet. FcIRL contained a predicted N-terminal acetylation site (M1-K5) and a NADPH-binding motif (G10-G-T-G13-Y-I-G16) in the N-terminal region, a conserved NmrA (nitrogen metabolite repression regulator) domain (V6-N244), multi-phosphorylation sites and one conserved N-glycosylation site (N214). Sequence homology comparison, phylogenetic analysis and advanced structures prediction all suggested that FcIRL belonged to the class of pinoresinol-lariciresinol reductase (PLR), which is a key enzyme in synthetic pathway of 8-8'-linked lignans, with function in catalyzing reduction of pinoresinol and lariciresinol into secoisolariciresinol, and medicinal secondary metabolism and resistance in F. cymosum.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Fagopyrum ; enzymology ; genetics ; Flavonoids ; genetics ; Lignans ; metabolism ; Molecular Sequence Data ; Oxidoreductases ; genetics ; Oxidoreductases Acting on CH-CH Group Donors ; genetics ; Protein Structure, Tertiary ; Sequence Homology, Amino Acid

Chondral injuries are short of self-healing ability and need to surgical repair after articular cartilage injury. Conventional treatment includes debridement and drainage under arthroscope, micro-fracture, osteochondral autograft transplantation (OATS), mosaiplasty and osteochondral allografts (OCA), autologous chondrocyte implantation (ACI). Debridement and drainage could remove pain factor, and has advantages of simple operation, wide clinical application and early clinical effect. Micro-fracture and osteochondral autograft transplantation is suitable for small area of cartilage repair, while the further effect showed that fibrous cartilage permeated by drill could decrease postoperative clinical effect. Osteochondral autograft transplantation has better advantages for reconstruction complete of wear-bearing joint. Autologous chondrocyte implantation and allogeneic cartilage transplantation are suitable for large area of cartilage defect, postoperative survival of allogeneic cartilage transplantation is effected by local rejection reaction and decrease further clinical effect. Cartilage tissue engineering technology could improve repair quality of autologous chondrocyte implantation, and make repair tissue close to transparent cartilage, but has limit to combined subchondral bone plate, reactive bone edema, bone loss and bad axis of lower limb. New technology is applied to cartilage injury, and has advantages of less trauma, simple operation, rapid recover, good clinical effect and less cost;and could be main method for treat cartilage injury with surgical repair technology. How to improve repair quality with compression resistance and abrasive resistance are expected to be solved.
Cartilage, Articular ; injuries ; surgery ; Chondrocytes ; transplantation ; Humans ; Knee Injuries ; surgery ; Knee Joint ; surgery ; Transplantation, Autologous