1.Role of zinc finger protein 1 in rat liver fibrosis and as related to TGFb expression.
Yujuan SHEN ; Yongping CHEN ; Zhenzhen LIU ; Xiaodong WANG ; Chunlei DAI ; Zhuo LIN
Chinese Journal of Hepatology 2014;22(4):285-288
OBJECTIVETo determine the role of zinc finger protein 1 (ZEB 1) in liver fibrosis and in regards to expression of the tumor growth factor-beta (TGFb) signaling factor using a rat model system.
METHODSSprague-Dawley rats were randomly divided into a normal (control) group, liver fibrosis (model) group and a liver fibrosis + therapy (ZEB1 intervention) group. The model group and the ZEB1 intervention group were given intraperitoneal injections of dimethylnitrosamine (DMN) for the first 3 days of each week over a 7-week period; starting at week 5, the ZEB 1 intervention group was started on a routine of every other day tail vein injections of recombinant ZEB1. During this 7-week period, the control group was given intraperitoneal injections of 0.9% NaC1 alone on the DMN schedule. Liver tissues were collected for pathological examination (with hematoxylin-eosin and Masson staining) and for detection of TGFb1 and ZEB 1 expression (by RT-PCR and western blotting). Measurement data were compared between groups using the single-factor analysis of variance test, followed by the least significant difference LSD test. Count data were analyzed by Fisher's exact test.
RESULTSThe model group's liver tissues showed degeneration and necrosis, as well as obvious fibrous septa accompanied by pseudo lobules. The ZEB 1 intervention group's liver tissues showed a significantly higher degree of fibrosis (x²=21.63, P=0), with more coarse fiber cords. The expression of ZEB1 and TGFb1 was significantly higher in the model group than in the control group (both P less than 0.05). However, the ZEB 1 intervention group showed the highest levels of ZEB 1 and TGFb1 expression (vs. model group, P less than 0.05).
CONCLUSIONZEB 1 may promote the development of liver fibrosis in rats through a mechanism involving the TGFb/Smad signaling pathway.
Animals ; Homeodomain Proteins ; pharmacology ; Liver ; drug effects ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Transcription Factors ; pharmacology ; Transforming Growth Factor beta1 ; metabolism ; Zinc Fingers
2.Application of Quality Control Circle in Shortening Drug Dispensing Time in the Inpatient Pharmacy of Our Hospital
Yujuan ZHUO ; Ning LUO ; Liu YANG ; Yanru QIAO ; Liezheng CAO ; Lian LI
China Pharmacy 2018;29(5):591-595
OBJECTIVE: To shorten drug dispensing time of inpatient pharmacy in our hospital, and to enhance the quality of pharmaceutical care.METHODS: The activity of quality control circle (QCC) was carried out by using "shorten drug dispensing time of inpatient pharmacy" as subject. According to the steps of QCC activity, key links and factors that influence the of drug dispensing time in the inpatient pharmacy were found out. Countermeasures were formulated and implemented according to practical reasons. The activity was evaluated by using the drug dispensing time in every 100 medical orders before and after QCC activity. RESULTS: According to the links of "dispensing medical order" and "checking medical order" that spent longest time, the countermeasures were put forward to improve work efficiency, such as rational drug distribution and design, opmitization of the dispensing route, realizing electronic large infusion prescription by strengthening system informatization, strengthening the management of confusing drugs, strengthening staff training and establishing performance appraisal system. The drug dispensing time was shortened from 43. 99 min (before QCC) to 21. 77 min (after QCC), and target achievement rate was 99. 24%. QCC staff showed positive growth in means, responsibility, team cohesion and other aspects. CONCLUSIONS: QCC activity can shorten the time of drug dispensing, improve work efficiency and pharmacist' s ability of solving problem, and help to improve pharmaceutical care in the inpatient pharmacy.
3.Comparison of oncology outcomes and anal function among laparoscopic partial, subtotal and total intersphincteric resection for low rectal cancers.
Bin ZHANG ; Quanlong LIU ; Yujuan ZHAO ; Guangzuan ZHUO ; Shuhui YIN ; Jun ZHU ; Ke ZHAO ; Jianhua DING
Chinese Journal of Gastrointestinal Surgery 2017;20(8):904-909
OBJECTIVETo compare the oncology outcomes and anal function among laparoscopic partial, subtotal and total intersphincteric resection(ISR) for low rectal cancers.
METHODSFrom June 2011 to February 2016, a total of 79 consecutive patients with low rectal cancers underwent laparoscopic ISR with hand-sewn coloanal anastomosis at our department. According to the distal tumor margin, partial ISR (internal sphincter resection at the dentate line) was used to treat tumors with distance <1 cm from the anal sphincter (n=28), subtotal ISR was adopted for the tumors locating between the dentate line and intersphincteric groove (n=34), and total ISR (resection at the dentate line) was applied in the treatment of intra-anal tumors (n=17). Anal function was evaluated by a standardized gastrointestinal questionnaire, Wexner incontinence score and Kirwan's classification. Metaphase oncological results and postoperative anal function were compared among three groups, and.
RESULTSOther than the distance of tumor low margin to dentate line (P=0.000) and serum CEA level (P=0.040), no significant differences were noted in baseline data among 3 groups (all P>0.05). The median follow up was 21(8-61) months. The 3-year disease-free survival rates in laparoscopic partial, subtotal and total ISR groups were 91.1%, 88.9%, 88.2% (P=0.901) and the 3-year local relapse-free survival rates were 91.1%, 72.9%, 80.2%(P=0.658), whose all differences were not significant. Thirty-eight patients who did not receive neoadjuvant chemoradiotherapy and underwent ileostomy closure for at least 24 months completed the evaluation of anal function, including 14 cases in partial group, 15 cases in subtotal group and 9 cases in total group. Of 38 patients, 73.7%(28/38) was classified as good function (Wexner incontinence score ≤10) and no patient adopted a colostomy because of severe fecal incontinence(Kirwan classification=grade 5). Furthermore, there were no significant differences in Wexner incontinence score and Kirwan classification among 3 groups (all P>0.05). However, patients with chronic anastomotic stoma stenosis showed worse anal function than those without stenosis [Wexner incontinence score: 18(9-20) vs 6(0-18), P=0.000; Kirwan grading: 3(2-4) vs. 2(1-4), P=0.002].
CONCLUSIONSAs the ultimate sphincter-saving technique, laparoscopic ISR can result in better oncologic outcomes and better anal function for patients with low rectal cancers. The different procedures of ISR may not affect the efficacy, but chronic anastomotic stoma stenosis deteriorates incontinence status.
4.Treatment of complications after laparoscopic intersphincteric resection for low rectal cancer.
Bin ZHANG ; Ke ZHAO ; Quanlong LIU ; Shuhui YIN ; Yujuan ZHAO ; Guangzuan ZHUO ; Yingying FENG ; Jun ZHU ; Jianhua DING
Chinese Journal of Gastrointestinal Surgery 2017;20(4):432-438
OBJECTIVETo summarize the perioperative and postoperative complications follow laparoscopic intersphincteric resection (LapISR) in the treatment of low rectal cancer and their management.
METHODSAn observational study was conducted in 73 consecutive patients who underwent LapISR for low rectal cancer between June 2011 and February 2016 in our hospital. The clinicopathological parameters, perioperative and postoperative complications, and clinical outcomes were collected from a prospectively maintained database. Perioperative and postoperative complications were defined as any complication occurring within or more than 3 months after the primary operation, respectively.
RESULTSForty-nine(67.1%) cases were male and 24(32.9%) were female with a median age of 61(25 to 79) years. The median distance from distal tumor margin to anal verge was 4.0(1.0 to 5.5) cm. The median operative time was 195 (120 to 360) min, median intra operative blood loss was 100 (20 to 300) ml, median number of harvested lymph nodes was 14(3 to 31) per case. All the patients underwent preventive terminal ileum loop stoma. No conversion or hospital mortality was presented. The R0 resection rate was 98.6% with totally negative distal resection margin. A total of 34 complication episodes were recorded in 21(28.8%) patients during perioperative period, and among which 20.6%(7/34) was grade III(-IIII( according to Dindo system. Anastomosis-associated morbidity (16.4%,12/73) was the most common after LapISR, including mucosa ischemia in 9 cases(12.3%), stricture in 7 cases (9.6%, 4 cases secondary to mucosa necrosis receiving anal dilation), grade A fistula in 3 cases (4.1%) receiving conservative treatment and necrosis in 1 case (1.4%) receiving permanent stoma. After a median follow up of 21(3 to 60) months, postoperative complications were recorded in 12 patients (16.4%) with 16 episodes, including anastomotic stenosis (8.2%), rectum segmental stricture (5.5%), ileus (2.7%), partial anastomotic dehiscence (1.4%), anastomotic fistula (1.4%), rectovaginal fistula (1.4%) and mucosal prolapse (1.4%). These patients received corresponding treatments, such as endoscopic transanal resection, anal dilation, enema, purgative, permanent stoma, etc. according to the lesions. Six patients (8.2%) required re-operation intervention due to postoperative complications.
CONCLUSIONAnastomosis-associated morbidity is the most common after LapISR in the treatment of low rectal cancer in perioperative and postoperative periods, which must be strictly managed with suitable methods.
Adult ; Aged ; Anal Canal ; surgery ; Anastomosis, Surgical ; adverse effects ; Blood Loss, Surgical ; statistics & numerical data ; Colectomy ; adverse effects ; Constriction, Pathologic ; etiology ; therapy ; Digestive System Surgical Procedures ; adverse effects ; Female ; Humans ; Ileostomy ; adverse effects ; Intestinal Mucosa ; pathology ; Ischemia ; etiology ; Laparoscopy ; adverse effects ; Lymph Node Excision ; statistics & numerical data ; Male ; Margins of Excision ; Middle Aged ; Necrosis ; etiology ; Operative Time ; Postoperative Complications ; etiology ; therapy ; Rectal Neoplasms ; complications ; surgery ; Rectovaginal Fistula ; etiology ; therapy ; Surgical Stomas ; Treatment Outcome
5.Risk factors for anastomotic leakage after laparoscopic intersphincteric resection for low-lying rectal cancer
Bin ZHANG ; Guangzuan ZHUO ; Yujuan ZHAO ; Ke ZHAO ; Yong ZHAO ; Jun ZHU ; Guowei NI ; Zhan CHEN ; Jianhua DING
Chinese Journal of General Surgery 2020;35(1):8-12
Objective To investigate the risk factors for anastomotic leakage (AL) after laparoscopic intersphincteric resection (Lap-ISR) for patients with low-lying rectal cancer.Methods This retrospective study was conducted in the Characteristic Medical Center of PLA Rocket Force from Jun 2011 to Nov 2018.151 patients undergoing Lap-ISR were enrolled for this study.Results All patients in this series had a defunctioning ileostomy.The overall leakage rate was 17.2% (26/151),including peri-operative AL (n =20) and delayed AL (n =6).In accordance with the grading system of the International Study Group of Rectal Cancer,there were 24 patients (15.9%) with AL Grade B (requiring active therapeutic intervention) and two patients (1.3%) with AL Grade C (requiring re-laparotomy).Univariate analysis showed that BMI (≥ 25 kg/m2),tumor annularity (≥ 3/4) and operation time (≥ 240 min) were associated with AL (P < 0.05).Multivariate analysis showed that operation time (≥ 240 min,OR =7.390,95% CI:2.483-21.988,P =0.000),tumor annularity (≥ 3/4,OR =6.233,95% CI:1.932-20.107,P=0.002) and higher BMI (≥ 25 kg/m2,OR=3.523,95% CI:1.275-9.738,P=0.015)were independently predictive of AL Conclusion Tumor annularity,operation time and higher BMI are independently associated with symptomatic AL after Lap-ISR.
6.Clinical and genetic analysis of 5 pediatric patients with hemiplegic migraine presenting as acute encephalopathy
Xiuwei ZHUO ; Shuhong REN ; Shuai GONG ; Weihua ZHANG ; Jiuwei LI ; Yujuan ZHANG ; Changhong DING
Chinese Journal of Pediatrics 2021;59(4):316-321
Objective:To analyze the clinical and genetic characteristics, diagnosis and treatment of hemiplegic migraine (HM) manifested as acute encephalopathy in children, so as to improve the understanding of this disease.Methods:The clinical data of 5 children diagnosed with HM characterized by acute encephalopathy who were admitted to Beijing Children′s Hospital affiliated to Capital Medical University from August 2018 to June 2020 were retrospectively analyzed.Results:Among the 5 cases, 3 were males and 2 females with an age of 9.7 (3.9-12.7) years. The age of disease onset was 7.0(2.1-12.7) years. The peak symptoms of 5 children showed encephalopathy such as drowsiness and coma, as well as other clinical manifestations including headache, visual abnormality, hemiplegia, aphasia, convulsions, and fever, etc. The time to reach the peak was on the 2nd-6th day of the course of the disease. Before the onset of the disease 2 cases were found to have mild brain trauma and 2 cases had similar attacks in the past. Brain magnetic resonance imaging (MRI) showed hemispheric or partial cerebral cortex swelling and restricted diffusion of subcortical white matter in all cases, and cerebellar atrophy in 3 cases. All children received symptomatic treatment, and 2 of them were also treated with low-dose corticosteroids in the meantime. Finally all cases recovered clinically from the attack, but one had atrophic changes left in the affected area on brain MRI. Whole exon sequencing revealed variations of CACNA1A gene in all cases, among which 4 were de novo mutations and 1 case inherited from the mother who had migraine without aura. After the diagnosis, the 5 children were treated with long-term flunarizine and followed up for 22(7-29) months by telephone or in the outpatient clinic. Before the last follow-up, none of them showed weakness or encephalopathy, but one still had intermittent headaches and occasional transient right limb numbness.Conclusions:Hemipleg is often accompanied by impaired consciousness in addition to headache, hemiplegia, aphasia, visual abnormality, etc. Most patients recover completely after a short period, while a few recover slowly and may suffer sequelae such as brain atrophy and cognitive impairment and even death. CACNA1A gene variation is the most common genetic variation. Flunarizine could prevent recurrence of severe attack.
7.Risk factors of coloanal anastomotic stricture after laparoscopic intersphincteric resection for low rectal cancer
Bin ZHANG ; Guangzuan ZHUO ; Lei TIAN ; Ke ZHAO ; Yong ZHAO ; Yujuan ZHAO ; Jun ZHU ; Tao ZHANG ; Jianhua DING
Chinese Journal of Gastrointestinal Surgery 2019;22(8):755-761
Objective To evaluate the risk factors of coloanal anastomotic stricture after laparoscopic intersphincteric resection (Lap?ISR) for patients with low rectal cancer. Methods A retrospective case?control study was performed to collect clinicopathological data from a prospective database (registration number: ChiCTR?ONC?15007506) at the Department of Colorectal Surgery, the Characteristic Medical center of PLA Rocket Force. From June 2011 to August 2018, a total of 144 consecutive patients with low rectal cancer who underwent Lap?ISR were enrolled in the study. Inclusion criteria: (1) reconstruction of digestive tract by end?to?end hand?made coloanal anastomosis (HCAA); (2) distance from lower tumor margin to anorected sphincter ring<1 cm and distance from lower tumor margin to intersphincteric groove ≥ 1 cm; (3) T1?3 stage tumor with expected negative circumferential resection margin evaluated by preoperative MRI or 3D endoanal ultrasound; (4) rectal cancer confirmed as well?or moderately ? differentiated adenocarcinoma; (5) preoperative Wexner incontinence score >10 points. Exclusion criteria: (1) follow?up period less than 3 months; (2) multiple primary cancers; (3) undergoing colonic J?pouch, coloplasty or reconstruction of end?to?side coloanal anastomosis; (4) death within perioperative period (within 3 months after surgery). Coloanal anastomotic stricture was diagnosed if the index finger or 12 mm electronic colonoscope had obvious resistance through the anastomosis or new rectum, or could not pass, accompanied by clinical symptoms such as difficult defecation and anal incontinence. Degree of anastomotic stricture was divided into 3 grades: grade A required anal enlargement, laxative or enema to assist defecation without active surgical treatment; grade B required surgery or endoscopic intervention; grade C required definitive ostomy, including unreducible preventive ileostomy or permanent colostomy. Univariate and multivariate analysis were used to evaluate the effects of 28 variables, including baseline data (age, gender, body mass index, neoadjuvant therapy, etc.), tumor?related factors (distance between tumor low margin and anal edge, maximum diameter of tumor, TNM staging, etc.), surgery?related factors (operation time, intraoperative blood loss, ISR procedure, anastomotic height, etc.) and anastomotic leakage, on the postoperative coloanal anastomotic stricture. Univariate analysis used χ2 test or Fisher′s exact test, then factors with P<0.05 were further included in multivariate analysis using logistic regression. Results A total of 144 patients were enrolled in the study, including 90 males and 54 females with a median age of 59 years and median BMI of 24.88 kg/m2. R0 resection rate was 96.5% (139/144). Median tumor distal resection margin was 1.5 (0.5 to 3.0) cm. Median follow?up was 31.5 (4 to 86) months. Coloanal anastomotic stricture was observed in 19 patients (13.2%), including 3 cases (2.1%) of grade A, 9 cases (6.2%) of grade B, and 7 cases (4.9%) of grade C. The median interval from the initial surgery to diagnosis of anastomotic stricture was 7 (1 to 31) months. Univariate analysis showed that male (χ2=6.795, P=0.009), radiotherapy (χ2=13.330, P=0.001), operation type of ISR (χ2=7.996, P=0.013), and anastomotic leakage (χ2=10.198, P=0.004) were associated with the postoperative coloanal anastomotic stricture. Multivariate analysis further indicated that male (OR=5.975, 95% CI:1.209?29.534, P=0.028), postoperative radiotherapy (OR=8.748, 95% CI: 2.397?31.929, P=0.001), and anastomotic leakage (OR=6.313, 95% CI: 1.834?21.734, P=0.003) were independent risk factor of postoperative coloanal anastomotic stricture. Conclusion For male patients, or patients with postoperative radiotherapy or anastomotic leakage, close follow?up should be carried out to prevent postoperative coloanal anastomotic stricture following Lap?ISR.
8.Risk factors of coloanal anastomotic stricture after laparoscopic intersphincteric resection for low rectal cancer
Bin ZHANG ; Guangzuan ZHUO ; Lei TIAN ; Ke ZHAO ; Yong ZHAO ; Yujuan ZHAO ; Jun ZHU ; Tao ZHANG ; Jianhua DING
Chinese Journal of Gastrointestinal Surgery 2019;22(8):755-761
Objective To evaluate the risk factors of coloanal anastomotic stricture after laparoscopic intersphincteric resection (Lap?ISR) for patients with low rectal cancer. Methods A retrospective case?control study was performed to collect clinicopathological data from a prospective database (registration number: ChiCTR?ONC?15007506) at the Department of Colorectal Surgery, the Characteristic Medical center of PLA Rocket Force. From June 2011 to August 2018, a total of 144 consecutive patients with low rectal cancer who underwent Lap?ISR were enrolled in the study. Inclusion criteria: (1) reconstruction of digestive tract by end?to?end hand?made coloanal anastomosis (HCAA); (2) distance from lower tumor margin to anorected sphincter ring<1 cm and distance from lower tumor margin to intersphincteric groove ≥ 1 cm; (3) T1?3 stage tumor with expected negative circumferential resection margin evaluated by preoperative MRI or 3D endoanal ultrasound; (4) rectal cancer confirmed as well?or moderately ? differentiated adenocarcinoma; (5) preoperative Wexner incontinence score >10 points. Exclusion criteria: (1) follow?up period less than 3 months; (2) multiple primary cancers; (3) undergoing colonic J?pouch, coloplasty or reconstruction of end?to?side coloanal anastomosis; (4) death within perioperative period (within 3 months after surgery). Coloanal anastomotic stricture was diagnosed if the index finger or 12 mm electronic colonoscope had obvious resistance through the anastomosis or new rectum, or could not pass, accompanied by clinical symptoms such as difficult defecation and anal incontinence. Degree of anastomotic stricture was divided into 3 grades: grade A required anal enlargement, laxative or enema to assist defecation without active surgical treatment; grade B required surgery or endoscopic intervention; grade C required definitive ostomy, including unreducible preventive ileostomy or permanent colostomy. Univariate and multivariate analysis were used to evaluate the effects of 28 variables, including baseline data (age, gender, body mass index, neoadjuvant therapy, etc.), tumor?related factors (distance between tumor low margin and anal edge, maximum diameter of tumor, TNM staging, etc.), surgery?related factors (operation time, intraoperative blood loss, ISR procedure, anastomotic height, etc.) and anastomotic leakage, on the postoperative coloanal anastomotic stricture. Univariate analysis used χ2 test or Fisher′s exact test, then factors with P<0.05 were further included in multivariate analysis using logistic regression. Results A total of 144 patients were enrolled in the study, including 90 males and 54 females with a median age of 59 years and median BMI of 24.88 kg/m2. R0 resection rate was 96.5% (139/144). Median tumor distal resection margin was 1.5 (0.5 to 3.0) cm. Median follow?up was 31.5 (4 to 86) months. Coloanal anastomotic stricture was observed in 19 patients (13.2%), including 3 cases (2.1%) of grade A, 9 cases (6.2%) of grade B, and 7 cases (4.9%) of grade C. The median interval from the initial surgery to diagnosis of anastomotic stricture was 7 (1 to 31) months. Univariate analysis showed that male (χ2=6.795, P=0.009), radiotherapy (χ2=13.330, P=0.001), operation type of ISR (χ2=7.996, P=0.013), and anastomotic leakage (χ2=10.198, P=0.004) were associated with the postoperative coloanal anastomotic stricture. Multivariate analysis further indicated that male (OR=5.975, 95% CI:1.209?29.534, P=0.028), postoperative radiotherapy (OR=8.748, 95% CI: 2.397?31.929, P=0.001), and anastomotic leakage (OR=6.313, 95% CI: 1.834?21.734, P=0.003) were independent risk factor of postoperative coloanal anastomotic stricture. Conclusion For male patients, or patients with postoperative radiotherapy or anastomotic leakage, close follow?up should be carried out to prevent postoperative coloanal anastomotic stricture following Lap?ISR.
9. Risk factors of coloanal anastomotic stricture after laparoscopic intersphincteric resection for low rectal cancer
Bin ZHANG ; Guangzuan ZHUO ; Lei TIAN ; Ke ZHAO ; Yong ZHAO ; Yujuan ZHAO ; Jun ZHU ; Tao ZHANG ; Jianhua DING
Chinese Journal of Gastrointestinal Surgery 2019;22(8):755-761
Objective:
To evaluate the risk factors of coloanal anastomotic stricture after laparoscopic intersphincteric resection (Lap-ISR) for patients with low rectal cancer.
Methods:
A retrospective case-control study was performed to collect clinicopathological data from a prospective database (registration number: ChiCTR-ONC-15007506) at the Department of Colorectal Surgery, the Characteristic Medical center of PLA Rocket Force. From June 2011 to August 2018, a total of 144 consecutive patients with low rectal cancer who underwent Lap-ISR were enrolled in the study. Inclusion criteria: (1) reconstruction of digestive tract by end-to-end hand-made coloanal anastomosis (HCAA); (2) distance from lower tumor margin to anorected sphincter ring < 1 cm and distance from lower tumor margin to intersphincteric groove ≥ 1 cm; (3) T1-3 stage tumor with expected negative circumferential resection margin evaluated by preoperative MRI or 3D endoanal ultrasound; (4) rectal cancer confirmed as well- or moderately-differentiated adenocarcinoma; (5) preoperative Wexner incontinence score >10 points. Exclusion criteria: (1) follow-up period less than 3 months; (2) multiple primary cancers; (3) undergoing colonic J-pouch, coloplasty or reconstruction of end-to-side coloanal anastomosis; (4) death within perioperative period (within 3 months after surgery). Coloanal anastomotic stricture was diagnosed if the index finger or 12 mm electronic colonoscope had obvious resistance through the anastomosis or new rectum, or could not pass, accompanied by clinical symptoms such as difficult defecation and anal incontinence. Degree of anastomotic stricture was divided into 3 grades: grade A required anal enlargement, laxative or enema to assist defecation without active surgical treatment; grade B required surgery or endoscopic intervention; grade C required definitive ostomy, including unreducible preventive ileostomy or permanent colostomy. Univariate and multivariate analysis were used to evaluate the effects of 28 variables, including baseline data (age, gender, body mass index, neoadjuvant therapy, etc.), tumor-related factors (distance between tumor low margin and anal edge, maximum diameter of tumor, TNM staging, etc.), surgery-related factors (operation time, intraoperative blood loss, ISR procedure, anastomotic height, etc.) and anastomotic leakage, on the postoperative coloanal anastomotic stricture. Univariate analysis used χ2 test or Fisher′s exact test, then factors with