Objective To understand pathogen spectrum of bacterial and fungal infection of central nervous system (CNS),and evaluate the etiological diagnostic value of universal primer polymerase chain reaction (PCR).Methods Data about patients with suspected or confirmed bacterial and fungal infection of CNS from January 2009 to March 2015 were collected,species of pathogens from cerebrospinal fluid (CSF)were analyzed,DNA from patients’CSF were performed PCR amplification and sequencing with universal primers of bacterial 16S rRNA and fungal 28S rRNA, PCR detection results were compared with CSF culture during the same period.Results A total of 400 patients were with confirmed or suspected bacterial or fungal infection of CNS,132 of whom were with positive CSF culture.150 pathogenic isolates were detected,including 48 isolates of gram-positive bacteria,90 gram-negative bacteria,and 12 fungi;the top three isolated bacteria were Acinetobacter baumannii (n =32 ),coagulase negative staphylococcus (n=16)and Klebsiella pneumoniae (n=13);the most common fungus was Cryptococcus neoformans (n=8).CSF from 88 infected patients and 20 non-infected patients were selected for PCR amplification,the sensitive of PCR am-plification assay was higher than the culture method (35.23% [31/88]vs 28.41 %[25/88],χ2 =4.17,P <0.05).

Objective To analyze the response rate and prognostic factors for patients with locally recurrent rectal cancer treated with hypofractionated chemoradiotherapy without reresection.Methods Totally 52 patients with locally recurrent rectal cancer received hypofractionated irradiation and concurrent chemotherapy from January 2006 to January 2013 were enrolled.All patients received intensity-modulated radiotherapy (IMRT).The median dose was 63.4 Gy (61.6-64.4 Gy) at 2.2-2.3 Gy/f,5 f/week.Thirteen patients underwent prophylactic irradiation at lymph nodes region,the total dose of 45-50.4 Gy with conventional fraction and a simultaneous integrated boost was used.All patients received concurrent chemotherapy,capecitabine at 1 650 mg·m-2 ·d-1,divided into 2 times,5 d/week.The variables were compared by the chi-square test or Fisher's exact test.Local control (LC) and overall survival (OS) were calculated with using the Kaplan-Meier method.Results For all patients,the clinical complete response (CR),partial response (PR),stable disease (SD) and progressive disease (PD) was 23.1％,38.5％,32.7％ and 5.8％,respectively.The response rate (CR + PR) for patients with previous irradiation to pelvis and without were 37.1％ and 71.1％,respectively (x2 =5.40,P ＜ 0.05);for patients with 1 and 2 or more recurrent subsites were 81.8％ and 46.7％,respectively (x2 =6.63,P ＜ 0.05).Acute grade 3 skin and hematologic toxicities occurred in 19 patients (36.5％) and 1 patient (1.9％),respectively.None occurred grade 4 toxicity and none occurred grade 3 or more gastrointestinal and urologic toxicities.Four patients showed severe late toxicity of anastomotic stricture and performed a stoma at transverse colon.No other severe late toxicities were observed.The LC at 5 years was 49.1％ and the OS was 23.1％.Conclusions For patients with locally recurrent rectal cancer,hypofractionated chemoradiotherapy without resection is an acceptable and effective regimen,the response rate and long-term outcomes are promising.

Objective To explore the clinical efficacy and safety of using simple arthroscopic operation combined with post-operative radiotherapy in the treatment of knee joint diffuse pigmented villonodular synovitis (DPVNS).Methods A total of 30 cases with knee DPVNS in Wuhan Central Hospital and General Hospital of Beijing Military Region from January 2009 to January 2014 were enrolled,including 18 males and 12 females with average age of 42.5 years (28-64 years).The average duration was 35.6 months(4-121 months).According to the decision of the patients,16 cases treated with operation combined with postoperative radiotherapy and the other 14 cases with operation alone.The treatment group accepted arthroscopic synovial excision and received local radiotherapy,the total dose of radiotherapy was 40 Gy,2 Gy/fraction.While the control group received arthroscopic operation alone.Patients were followed up,which the average time of 38.7 months (12-72 months).The recurrence rate,clinical KSS score and function score were compared between the two groups.Results The efficiency of treatment group (14/16) was significantly higher than the control group (11/14) (x2 =9.87,P ＜ 0.05),and the recurrence rate of the former(1/16) was significantly lower than that latter (3/14) (x2 =1.83,P ＜ 0.05).Furthermore the KSS clinical score and function score were improved significantly in the treatment group(x2 =15.00,15.78,P ＜ 0.05).The main side effect of radiation therapy were leucopenia,limb edema,skin pigmentation and so on,meanwhile the treatment group was well tolerated.Conclusions Arthroscopic operation combined with postoperative radiotherapy is safe and effective in treatment of DPVNS,and it is worthy of clinical application.

Objective To determine the margins of planning target volume (MPTV) in primary cervical cancer patients with tomotherapy and evaluate the importance of automatic registration(AR) plus manual registration.Methods The setup errors of 29 primary cervical cancer patients receiving external radiation from June 2012 to Dec 2014 were measured by megavoltage computed tomography (MVCT),which were performed at least two times weekly before treatment and were registered with the planning CT.The setup errors between automatic registration and total shift (TS) including both AR and manual registration were compared MPTV was calculated.Results Setup errors were collecte from 443 sets of MVCT in 29 patients.AR and total shift (TS) values in the x,y,z directions and rotation angle were (-0.9±2.3),(0.0±3.1),(1.0±2.6) mm,0.2° ±0.8° and (-0.8±1.8),(-0.4±3.4),(l.4 ± 2.5) mm,0.1° ± 0.5°,respectively.There were statistically significant differences between the two groups in all directions except for the x axis (t =5.1,-5.2,3.2,P ＜ 0.05).MPTV were 4.6,5.7,3.3 mm in the x,y,z directions,respectively.Conclusions Manual registration is necessary after automatic registration in cervical cancer patients with tomotherapy.For patients with cervical cancer treated by tomotherapy,planning target volume MPTV parameters are suggested to be 5,6,4 mm in the x,y,z directions.

Objective To evaluate the value of amplification and sequencing of 16S rRNA gene in the identification of clinical rare pathogenic bacteria,and guide the diagnosis and treatment for related clinical infection.Methods 12 bacterial isolates that were difficult,or unable to be identified with conventional laboratory methods,or with special phenotypes were collected. The 16S rRNA gene was amplified by polymerase chain reaction (PCR),then sequenced for identifying bacterial species through BLAST comparison,clinical characteristics of related infection were ana-lyzed.Results 12 clinical isolates were all positive for PCR amplification (about 1500 bp),species were all identi-fied (similarity≥99% ),the identified strains were Listeriamonocytogenes(n= 2),Brucellamelitensis(n= 2),Fu-sobacteriummortiferum(n= 1),Rothiaaeria(n= 1),Nocardiafarcinica(n= 1),Staphylococcussaccharolyticus (n= 1 ),Rhizobiumradiobacter(n= 1 ),Prevotellabivia(n= 1 ),Ralstoniamannitolilytica(n= 1 ),and Atopobium vaginae(n= 1 ). The sensitivity of 16S rRNA gene amplification was high,and the minimum detection limit of Escherichiacoli ATCC 25922 was 1.5×101 CFU/mL. Clinical data of 12 patients revealed that these strains can cause multi-sites and multi-types of infection,after patients received targeted antimicrobial therapy,11 improved, and 1 died.Conclusion Sequencing for 16S rRNA gene can rapidly and accurately identify rare,anaerobic,and difficult cultured bacteria,provide laboratory evidence for etiological diagnosis and treatment of different types of infection.

Objective To observe the interfractional variation and actual dose for cervical cancer patients treated with tomotherapy.Methods Five patients who received tomotherapy were chosen from Aug 2013 to Feb 2014.A megavohage computed tomography (MVCT) scan was performed before treatment and then registered with the planning CT images.Dose distributions were recalculated and targets were contoured on the MVCT images.The differences between the actual radiation and planning were analyzed.Results In the patients received external radiotherapy, the decline in cervix volume and maximum diameter was 68.90％ and 26.91％ , respectively (t =5.21, 8.39, P ＜0.05).Cervix, uterus and CTV movement in left-right, anteroposterior, superoinferior were 1.43,-7.72, 0.02,-0.40,-1.24, -6.51,-0.43,-1.68and-0.22mm.The medianCTV V95％ was 99.40％ (95.96％-100％), and missing volume was 6.94 cm3 (0-32.30 cm3).Conclusions During radiotherapy for cervical cancer patients, the volume, position and doses are different between initial plan and actual radiation.Based on image guided radiation therapy (IGRT), missing targets are limited.

Objective To compare acute toxicity for stage Ⅱ-Ⅲ patients with rectal cancer irradiated with helical tomotherapy (HT) and conventional five-field intensity-modulated radiotherapy (5-IMRT).Methods The data of 84 stage Ⅱ-Ⅲ patients with rectal adenocarcinoma treated with concurrent chemoradiotherapy (CRT) were retrospectively analyzed.19 patients underwent postoperative CRT,and 65 patients underwent preoperative CRT.43 patients received radiotherapy with HT and 41 patients with 5-IMRT.The delineation on clinical target volume (CTV) and gross tumor target (GTV) was similar for two groups.The CTV to plan tumor volume (PTV) margins were 1.0 cm for patients with 5-1MRT and 0.5 cm for patients with HT.For all patients,a dose of 45.0-50.4 Gy,in daily fractions of 1.8 Gy,was delivered to PTV.For 45 patients with high risk factors,simultaneous integrated boost (SIB) was given to the tumor or tumor bed of a total dose of 55.0-60.0 Gy,in daily fractions of 2.1-2.3 Gy.Before treatment,the patients treated with HT underwent scanning by the tomotherapy-integrated megavoltage computed tomography (MVCT) scan modality and were positioned by co-registration of these images to the original kilovoltage planning CT image set.Concurrent capecitabine every day 1 600 mg/m2,twice daily on every day in the week.Results The rates of grade ≥ 2 acute cystitis were 7.0 ％ (3 cases) in HT group and 2.4 ％ (1 case) in 5-IMRT group (P =0.616),and ≥3 grade acute diarrhea were 4.7 ％ (2 cases) and 12.2 ％ (5 cases),respectively (P =0.259).≥2 grade leukopenia were 48.8 ％ (21 cases) and 19.5 ％ (8 cases),respectively (P =0.005),≥1 grade anemia were 34.9 ％ (15 cases) and 14.6 ％ (6 cases),respectively (P =0.032),and ≥1 grade thrombocytopenia were 23.3 ％ (10 cases) and 14.6 ％ (6 cases),respectively (P =0.314).Conclusions There is no significant difference in acute diarrhea and cystitis for patients treated with HT and 5-IMRT.Leukopenia and anemia in patients treated with HT are worse than those in patients with 5-IMRT,and thrombocytopenia is similar in the two groups.

Objective:To evaluate the effect of radical image-guided radiotherapy (IGRT) on the target dose in cervical cancer and investigate the appropriate application mode.Methods:Twenty patients with cervical cancer treated with helical tomotherapy (HT) in Seventh Medical Center of PLA General Hospital from 2012 to 2016 were selected. A megavoltage CT (MVCT) scan was performed before each treatment. The obtained MVCT images were used for dose parameter in the adaptive module of HT to obtain the actual dose (Plan 1) and the non-image-guided dose parameter was simulated (Plan 2). Each single dose distribution and the corresponding fused CT images were transferred to the software Mimvista 6.5 to obtain the total radiation dose parameter by dose superposition.Results:The motion of CTV, uterus and GTV in Plan 2 was significantly larger than that of Plan 1(all P<0.05), and the largest changes were seen in the ventrodorsal and uterine direction. The V 45Gy, V 50Gy, D 98% and D mean of CTV and uterus and V 50Gy of GTV in Plan 2 were significantly decreased compared with those in Plan 1(all P<0.05). The left-right motion of Plan 1 was negatively correlated with D 2% and D 98% of CTV and uterus (both P<0.05). The head-foot motion was negatively associated with V 45Gy and V 50Gy of GTV (both P<0.05). The ventrodorsal motion was negatively correlated with D 98% of uterus ( P<0.05). The left-right motion of Plan 2 was negatively correlated with D 2% of CTV and V 50Gy of uterus (both P<0.05). The head-foot motion was negatively associated with D 98% of CTV, and D 98%, D mean, V 45Gy and V 50Gy of uterus (all P<0.05). The ventrodorsal motion was negatively correlated with D 98% of CTV, D 98%, D mean, V 45Gy and V 50Gy of uterus, and D mean and V 45Gy of GTV (all P<0.05). Conclusions:In intensity-modulated radiotherapy for cervical cancer, the uterine body displacement is large and the low CTV area is mainly located in the uterine body. IGRT can significantly reduce the dosimetric deviation induced by organ movement.

PURPOSE: Lung adenocarcinoma (LA) is one of the major types of lung cancer. MicroRNAs (miRNAs) play an essential role in regulating responses of natural killer (NK) cells to cancer malignancy. However, the mechanism of miR-218-5p involved in the killing effect of NK cells to LA cells remains poorly understood. MATERIALS AND METHODS: The expression of miR-218-5p was examined by quantitative real-time polymerase chain reaction (qRT-PCR). Serine hydroxymethyl transferase 1 (SHMT1) level was detected by qRT-PCR or western blots. Cytokines production of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) were detected by ELISA. The killing effect of NK cells to LA cells was investigated using lactate dehydrogenase cytotoxicity assay kit. The interaction of miR-218-5p and SHMT1 was probed by luciferase activity assay. Xenograft model was established to investigate the killing effect of NK cells in vivo. RESULTS: miR-218-5p was enhanced and SHMT1 was inhibited in NK cells of LA patients, whereas stimulation of interleukin-2 (IL-2) reversed their abundances. Addition of miR-218-5p reduced IL-2-induced cytokines expression and cytotoxicity in NK-92 against LA cells. Moreover, SHMT1 was negatively regulated by miR-218-5p and attenuated miR-218-5p-mediated effect on cytotoxicity, IFN-γ and TNF-α secretion in IL-2-activated NK cells. In addition, miR-218-5p exhaustion inhibited tumor growth by promoting killing effect of NK cells. CONCLUSION: miR-218-5p suppresses the killing effect of NK cells to LA cells by targeting SHMT1, providing a potential target for LA treatment by ameliorating NK cells function.
Adenocarcinoma ; Blotting, Western ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Heterografts ; Homicide ; Humans ; Interleukin-2 ; Killer Cells, Natural ; L-Lactate Dehydrogenase ; Luciferases ; Lung Neoplasms ; Lung ; MicroRNAs ; Necrosis ; Real-Time Polymerase Chain Reaction ; Serine ; Transferases

OBJECTIVE To evaluate the quality of total flavonoids of Lamiophlomis rotata from different producing areas. METHODS Total flavonoids of S1-S15 batches of L. rotata from different producing areas were extracted by percolation and purified by polyamide column. The content of total flavonoids was determined by UV spectrophotometry， and its purity was calculated. HPLC fingerprint chromatograms and control fingerprint of total flavonoids from 15 batches of L. rotata were established with Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 version）. Their similarities were analyzed. Cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis were used to evaluate the quality of total flavonoids from 15 batches of L. rotata， and the main components which affected the quality of total flavonoids were analyzed. RESULTS The purities of total flavonoids from 15 batches of L. rotata were 55.82%-94.12%， with an average value of 77.72%； a total of five common peaks were identified in the fingerprint， and No. 3 peak was luteolin； the similarities between the fingerprint of each batch of samples and the control fingerprint were 0.925-1.000. By cluster analysis， S1 and S3-S9 were clustered into the first class， which were samples from Qinghai Province and Tibet Autonomous Region. S14 and S15 were clustered into the second class， which were samples from Yunnan Province. S10-S13 were clustered into the third class， which were all samples from Sichuan Province. S2 was clustered into the fourth class. The principal component analysis showed that the qualities of samples from the first and fourth classes were better； peaks 2， 3 and 5 were identified as the main components that caused the differences among different batches of samples by orthogonal partial least squares-discriminant analysis. CONCLUSIONS The qualities of total flavonoids of L. rotata from Qinghai Province and Tibet Autonomous Region are better.